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Candiolo Cancer Institute Fondazione del Piemonte per l’Oncologia

Istituto di Ricovero e Cura a Carattere Scientifico

Scientific Report 2015 - 2016

www.ircc.it

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Scientific Report 2015 - 2016

Contents

About Us 5

Mission 7

Research Synopsis 11

Mechanisms of Cancer Onset and Progression 13

From Molecular Biology to “Precision Medicine” 31

Investigational Clinical Oncology 53

Applied Clinical Research 59

Core Facilities 81

Clinical Services 83

Educational Activities 87

Grant Office and Research Administration 89

Electronic Data Processing Unit - Ethics Committee 90

List of Publications 91

4 | About Us

The Candiolo Cancer Institute

About Us | 5

THE CANDIOLO CANCER INSTITUTE - IRCCS

The Candiolo Cancer Institute is a private non-profit institution founded and supported by the Fondazione Piemontese

per la Ricerca sul Cancro-Onlus (FPRC) and operated by the Fondazione del Piemonte per l’Oncologia (FPO: co-founded

by FPRC and the Regione Piemonte). The Institute is a recognized IRCCS (Istituto di Ricovero e Cura a Carattere

Scientifico) by the Italian Ministry of Health. The Candiolo Cancer Institute works in synergy with the University of

Torino Medical School. Its mission is a significant contribution to fight cancer, by understanding the basics, and by

providing state-of-the-art diagnostic and therapeutic services. The interface between molecular biology and medice is

at the core of the Institute. FPRC provides enduring fund raising to complete and develop the Institute’s buildings and

technologies in order to foster research.

CANDIOLO CANCER INSTITUTE

6 | Mission

Donna Allegra Agnelli

President

Fondazione Piemontese

per la Ricesca sul Cancro

ONLUS (FPRC)

Marco Boglione

President

Fondazione del Piemonte

per l’Oncologia (FPO)

Giampiero Gabotto

CEO

(FPRC - FPO)

Mission| 7

MISSION

The Candiolo Cancer Institute is a biomedical and clinical research center entirely

devoted to the study and the treatment of cancer. Its mission is to transfer experi-

mental preclinical information into clinical practice, through the continuous flow of

knowledge from the fields of genetics, molecular and cell biology and pathology.

The Institute aims to offer a significant contribution to the defeat of the disease

through scientific research and clinical practice of excellence. In order to fulfill

these goals, the Institute: (i) capitalizes on knowledge by conducting scientific rese-

arch in oncology and – at the same time - by promoting fast transfer of knowledge

to the clinical practice; (ii) provides assistance in cancer prevention, including the

identification of genetic risk factors; (iii) performs diagnostic studies, using state-

of-the-art instrumentation and technology; (iv) provides a full cycle of treatment on

the premises, employing - besides the best conventional therapies - protocols for

novel targeted therapies and clinical trials for ‘precision medicine’.

Focus

Clinical and basic research makes the Institute of Candiolo a center of excellence focussed on the study, pre-

vention, and treatment of the dreadful complication of cancer, metastasis.

Molecular diagnosis and “precision” medicine

The success of targeted therapies is based on the rationale that the target molecule is ‘druggable’ - as a conse-

quence of the genetic anomaly - in the tumor but not in healthy tissues. Therefore the target molecule deactiva-

tion has consequences restricted to the neoplastic mass, with a minimum of ‘off-target’ consequences, leading

to generic organ damage. This notion has two important clinical outcomes: first of all, before treating patients

with a given targeted therapy, it is necessary to verify the presence of the genetic lesion “predictive” of sensiti-

vity to the drug. Second - in the perspective of targeted therapies - tumors will be classified not only according

to their site of origin and/or morphological features, but also by the molecular lesion(s) which earmark them

and, at the same time, make them vulnerable to a targeted treatment. Therefore, new therapies are not only

“targeted” but also “personalized”. This new approach is called “precision medicine”.

Clinical Research

At the Institute, oncologists, surgeons and radiotherapists cooperate with scientists to design clinical trials based on

molecular data, intended to verify hypotheses and generate novel ones. In order to make this cooperation productive,

the Institute manages a daily net of interactions involving formal aspects (seminars and meetings), training (re-

fresher courses and lessons), and operational efforts (contacts with pharmaceutical companies, and management

of regulatory instruments). Clinical Research is the last and more direct haven to improve the assistance to cancer

patients, providing them with the most appropriate, novel, safe and effective therapeutic approach, in accordance

with the genetic profile of their own tumor, as an ultimate means of increasing their life expectancy.

Prof. Paolo M.Comoglio MDScientific Director

8 | Mission

Investigational Clinical Oncology (INCO)

Cancer is a complex disease, tied to genetic lesions that increase in number over time, as a consequence of

genetic instability and exposure to environmental carcinogens. Thus, a cell clone proliferates and invades the

adjacent tissues without control. Cancer cells show several genetic anomalies that tend to increase as time

passes. Cancer’s heterogeneity makes it a hard-to-attack desease because of its multiple and continuously

changing target. The molecular lesions that cause and sustain most tumors are, however, finite in number (the

bona fide oncogenes). These findings have driven clinical pharmacology to commit to an epochal effort to cre-

ate drugs, called “targeted”, able to contrast the function of specific oncogenes. The strategy of the Institute

at Candiolo intends to make significant contributions to the field of cancer targeted therapy by: (i) identifying

pathologies and recruiting patients sensitive to the therapies currently in use; (ii) planning and performing - in

international networks - the related clinical trials; (iii) developing translational and preclinical research aimed

at designing new targeted therapies.

Translational Research (ECMO: Experimental Clinical Molecular Oncology)

Translational research is the “heart” of the Institute, bridging the gap between basic and clinical research. Ba-

sed on recent technological advances, genomic analysis makes possible –in a significant percentage of cases–

the identification of genetic alteration(s) with a “driver” role in tumor development. However, the contribution

of each lesion to the transformed phenotype remains elusive. Moreover, there is still insufficient knowledge of

the mechanisms that control the lack of response to targeted therapies, even in the presence of the molecular

target (primary resistance), as well as of the mechanisms that lead to a progressive attenuation of the response

after prolonged treatment (acquired resistance). ECMO research aims at the fulfillment of some ambitious Go-

als to integrate the traditional prognostic and diagnostic factors with a detailed characterization of the genetic

and functional alterations of the tumor; to identify new malfunctioning regulatory pathways in cancer; to isolate

and study cancer stem cells; to develop new preclinical platforms that can reliably disclose - and understand

in detail– the prospective results of clinical practice. The generation of this kind of knowledge is necessary to

design clinical trials that will no longer be based on empirical observations but on a strong rationale.

Basic Research

The current knowledge of the mechanisms of cancer onset and progression is provided by basic disciplines,

such as genetics, cell and developmental biology. Thanks to these studies, it is possible to classify tumors not

only according to the basis of their site of origin and histopathological features, but, notably, according to

the identification of the genetic lesion(s) that support their growth. Basic Research tasks at the Institute are

aimed at understanding the mechanisms that control normal cell functions responsible for proliferation, and

at analyzing how these mechanisms are corrupted during neoplastic transformation. The topics include signal

transduction, DNA duplication, cell division, differentiation, senescence, apoptosis and cell motility. Recent

studies suggest that anti-neoplastic therapy is really effective not only when it hits the appropriate molecular

target (vide infra), but especially when it hits the cells feeding the tumor mass. Indeed, most of the cells of the

neoplastic mass are quite innocuous and can be attacked with classical therapies, while only a small fraction

of them are resistant to treatment and able to regenerate the tumor. This small subpopulation includes the

“cancer stem cells” – strictly related to normal “stem cells” – which control the development of our organism

during embryonic life and allow us to renew worn parts in adult life. The up-to-date conceptualization of Cancer

defines it as a “somatic, genetic disease of the stem cells”.

Paolo M.Comoglio MD (Scientific Director)

Mission | 9

TWENTY YEARS OF RESEARCH AT THE INSTITUTEA story of courage and success

In the fall of 1996, even before the official opening, the most popular Turin daily newspaper dedicated a series

of full-page ads to the brand-new Candiolo Cancer Institute. It was a truly innovative advertising campaign in

terms of communication: big photos taking up the entire page and a short catch phrase at the bottom. One

of the most incisive pages recited something like, “In Candiolo, the light never goes out” and the photo captu-

red the bright windows on the fourth floor of the west wing, on an ordinary night, rather late. That lab on the

fourth floor had been installed in June of that year and was directed by a professor in his 50s, with a solid and

successful research experience behind him and ambitious ideas for the future. He could count on the work of a

dozen young researchers, as ambitious as he was.

Twenty years have passed since then. The lab on the fourth floor does no longer exist: in the summer of 2015, it

was relocated to the three large floors of the new research tower; the director of the small lab has become the

scientific director of the Institute; the young researchers have reached their 50s (some more, some less) and

have forged their ways within the international scientific community with brilliant discoveries and successes.

Meanwhile, new research labs were opened up and new scientists were recruited (often from abroad), giving

their precious contribution to the image and the visibility of the Institute with their own discoveries and succes-

ses. Young generations have been formed and are now about to take flight. Over the years, serial installments

of state-of-the-art technologies have rapidly grown old – as it often happens with this kind of instruments – but

have been promptly replaced with the latest models available. Research has developed and modified year after

year, answering and following the needs of the scientific community: the study of the invasion processes and

metastases, and of the mechanisms that generate new tumor vessels, first evolved into the exploration of the

molecular bases underlying response to non-chemotherapy targeted drugs, and then into the analysis of how

the tumors that initially respond to therapy become resistant after a certain period of treatment.

At the same time, the Institute has grown in terms of clinical assistance: in rapid sequence in-patient bed

areas, the ORs, the Outpatient Surgery and Oncology Units, and the Pathology and Clinical Biochemistry Divi-

sions were opened; the patients were ever increasing; the most advanced instrumentation (and often the only

one available in the whole Italian landscape) for diagnosis and therapy was set up. Over time, researchers and

clinicians started talking each other and working together, finding a common language to identify shared goals.

On the one hand, researchers acknowledged the unmet medical needs that had to be addressed by experimen-

tal approaches. On the other hand, clinicians recognized the value of molecular analysis to inform diagnostic

and therapeutic approaches. As a reward for all these efforts, in 2013, along came the acknowledgment of the

Institute as a Comprehensive Research Center (IRCCS). Candiolo is now one of the few centers of excellence

meticulously evaluated by the Italian Ministry of Health, which monitors the fulfilment of extremely high stan-

dards of scientific production and healthcare.

The lights of Candiolo are still on when it’s dark outside. Sometimes they faded, when the night was falling,

to honor the rest of those who lost the battle; but over these twenty years, they have shone on the all-watched

nights of the researchers performing their long experiments and on the night shifts of doctors and nurses, and

have brightened the way home for the many who made it through.

Livio Trusolino MD

10 | Mission

1996, The Pioneers

Research Synopsis | 11

RESEARCH SYNOPSIS

Topic 1: The mechanisms of onset and progression of cancer1.1 Role of Semaphorins in invasion, metastasis and angiogenesis1.2 The plasma membrane in cell migration and invasiveness1.3 Tumor angiogenesis Topic 2: From Molecular Biology to ‘Precision Medicine’2.1 Oncogenes and growth factors 2.2 Genetics of the response to anti-cancer therapy2.3 Preclinical models of ‘personalized’ therapies2.4 Resistance to targeted therapy Topic 3: Investigative clinical research: rationale for the planning of clinical trials3.1 HERACLES and ARES: ‘Precision medicine’ trials for metastatic colon carcinoma3.2 HERLAP: ‘Personalized’ treatment of mammary carcinoma3.3 AGNOSTOS: Therapy of metastatic cancers of unknown primary origin Topic 4: Applied clinical research4.1 Colorectal cancer: onset and progression4.2 Implementation of current therapeutic strategies4.3 Novel approaches to surgical oncology4.4 Radiotherapy, imaging and laboratory medicine

12 | The mechanisms of cancer onset and progression

The cell ’scattering’: a complex biological phenomenon resulting from

activation of a cascade of events triggered by the MET oncogene

The mechanisms of cancer onset and progression | 13

Research Topic 1: Mechanisms of cancer onset and progression

The current knowledge of mechanisms of cancer onset and progression has been provided by basic disciplines, such as cell biology, developmental biology and genetics. The tasks of Research Topic 1 are aimed at understanding the mechanisms controlling the normal cell functions responsible for the regulation of cell proliferation and at analyzing how these mechanisms are corrupted during neoplastic transformation. The topics include signal transduction, DNA duplication and cell division processes, differentiation, senescence, apoptosis and cell invasion. Special attention is given to the microenvironment and to the relations of the neoplastic cell with adjacent cells. The angiogenic process and the vascularization of the tumor mass have been studied in detail in the past.

Research Topic 1 is focussed on:1.1 The role of Semaphorins in invasiveness, metastases and angiogenesis1.2 The plasma membrane in metastasis dissemination1.3 Tumor angiogenesis


Semaphorins: structure & functionsThe elucidation of the functional role and molecular mechanisms through which Semaphorin genes control the invasiveness of tumor cells and the formation of metastasis will prompt the development of innovative diagnostic and therapeutic approaches

TeamMichael Rehman, Sabrina Rizzolio, Lorena Capparuccia, Chiara Battistini, Gabriella Cagnoni, Gurrapu Sreeharsha, Massimo Accardo

Research topic:

Semaphorins and Semaphorin Receptors in Cancer Progression.

Background:

Semaphorins and their receptors (Neuropilins and Plexins), beyond their role in embryo development and

morphogenesis, are relevant players in cancer progression and emerging targets for innovative therapeutic ap-

proaches. Semaphorins form a conserved family of over 20 members in vertebrates, including transmembrane

molecules and secreted members. Secreted semaphorins often act to restrict cell migration; for instance, they

can negatively regulate angiogenesis, preventing its pathological role in cancer. Certain semaphorins can play

diverse functional roles in different cell populations, due to the involvement of distinctive receptor complexes

and signaling cascades. Our lab demonstrated that Sema3A and Sema3E are relevant targets for innovative

therapeutic approaches, as they can regulate cancer cells and stromal cells in concomitant manner in the tu-

mor microenvironment. In addition to their role as semaphorin and VEGF receptors, we and others found that

neuropilins represent receptor hubs on the cell surface, controlling the function of multiple signaling cascades

that support tumor growth, such as EGFR and integrins.

Research achievements:

The laboratory has been working on several aspects of the research topic. We studied the function of Plexin-D1

and Neuropilin-1 receptors in cancer cells and in the tumor microenvironment, and found that they can play

multiple roles in cancer progression. For instance, oncogene addicted tumor cells use Nrp1 to uphold survival

signaling cascades in response to targeted therapies. Moreover, Nrp1 mediates Sema3A-dependent recruitment

and localization of tumor associated macrophages in hypoxic tumor areas, where they can deploy their pro-

angiogenic and immune-suppressor activity. Beside secreted family members, transmembrane semaphorins

are also currently investigated in the lab, such as Sema4C and Sema6A. We accomplished novel and exciting

results about their role in cancer cells, regulating their intrinsic metastatic potential and therapeutic response

to targeted therapies. The lab is also a leader in semaphorin research at international level, contributing expert

know-how and reagents to frontline collaborative studies.

Conclusions and perspectives:

Semaphorins and their receptors are novel and exciting players in cancer progression. By understanding the

specific role of different family members in human cancers, we could validate novel predictive markers of

progression or response to innovative targeted therapies. Moreover, certain semaphorins (and receptors) have

already been put forward as potential targets for functional interference approaches, aimed at regulating angio-

genesis, cancer cell invasiveness, and metastatic progression.

Selected references


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Luca Tamagnone MDLaboratory of Cancer Cell Biology

Associate ProfessorUniversity of Torino Medical School

E-mail: [email protected] Phone. +39.011.9933204

Fax. +39.011.9933225

Battistini C, Tamagnone L. Transmembrane semaphorins, forward and reverse signaling: have a look both ways.Cell Mol Life Sci. 2016 73:1609-1622

Serini G, Tamagnone L. Bad vessels beware! Semaphorins will sort you out! EMBO Mol Med. 2015 26;7:1251

Cagnoni G, Tamagnone L. Semaphorin receptors meet receptor tyrosine kinases on the way of tumor progression. Oncogene 2014, 33:4795-802

Biographical Sketch:

Luca Tamagnone, MD, PhD started his research activity with Paolo Comoglio. During his training in the lab of

Kari Alitalo, he discovered new genes encoding tyrosine kinases, such as RYK and BMX (published in Oncogene,

1993 and 1994). Returning to Italy, Dr. Tamagnone started working on a new family of receptors, the Plexins

(described in PNAS-USA, 1996). Three years later he published the identification of plexin ligands, the Se-

maphorins (in Cell, 1999). As tenured Associate Professor at the University of Torino, and Young Investigator of

the European Molecular Biology Organization, in 2001, Dr. Tamagnone started his own lab in the Institute for

Cancer Research of Candiolo-Torino (IRCC). He focused his attention on semaphorin/plexin activities in cancer

cells, identifying fundamental signaling mechanisms and functional properties in the regulation of tumor pro-

gression (e.g. Neuron, 2001; Nat Cell Biol, 2002; Nature, 2003; FASEB J, 2004; J Cell Sci, 2005). By exploiting

experimental models in vivo, he found that semaphorin signals regulate multiple steps of tumor progression:

from tumor growth, to angiogenesis, to the recruitment of tumor-associated leucocytes, to the formation of

distant metastases (e.g. J Exp Med, 2008; J Clin Invest, 2010; EMBO Mol Med, 2012), validating their relevan-

ce as targets for molecular therapy. Moreover, recent findings have revealed novel functions of the semaphorin

co-receptor Neuropilin-1 in the regulation of EGFR oncogene signalling (Cancer Res, 2012), as well as in the

recruitment of tumor associated macrophages (Cancer Cell, 2013).


Dynamic control of cell adhesion: ‘normalization’ of tumor blood vesselsThe pharmacological modulation of integrin function can be therapeutically exploited to improve the biodistribution of anti-neoplastic drugs and counteract the hypoxia-driven metastatic dissemination of cancer cells

TeamDonatella Valdembri, Chiara Camillo, Noemi Gioelli, Giulia Mana, Chiara Sandri, Giulia Villari

Research topic:

Grasping the molecular basis of cell adhesion dynamics to design effective ‘vascular normalizing’ drugs for anti-

cancer therapy.

Background:

Tumor blood vessels are structurally and functionally aberrant, thus they hamper the delivery of anti-cancer drugs and

cause hypoxia-driven metastatic dissemination. Normalizing the vasculature of tumors could hence sizably improve

anti-cancer therapy. The binding of vascular endothelial cells (ECs) to the extracellular matrix (ECM) is mediated by

integrins, a class of adhesive receptors that can assume active or inactive conformations, respectively characterized

by high or low affinity binding to ECM ligands. Blood fluid shear stress is the main determinant of normal vascular

architecture and function, its foremost outcomes being that of triggering integrin activation and fostering EC-to-ECM

adhesion. As a direct consequence, ECs modify their adhesive interactions and reciprocal positions in space, finally gi-

ving rise to a remodeled, mature, and functional vascular tree. In this framework, inhibitors of integrin function should

warrant a swift responsivity of EC-to-ECM adhesion to variations in blood flow-elicited forces. Therefore, pharmacologi-

cal modulation of integrin function might be therapeutically exploited to ‘normalize’ the tumor vasculature.


We have previously shown how: (i) the chemorepulsive guidance cue semaphorin 3A (Sema3A) and its receptors

plexins and neuropilin-1 (Nrp1) allow correct vascular morphogenesis by negatively modulating the conformational

activation of integrins and inducing active/ECM bound integrin endocytosis; (ii) the abrogation of Sema3A-depen-

dent integrin-inhibitory signals underpins the alterations that characterize tumor blood vessels; (iii) somatic gene

transfer of Sema3A restores the physiological inhibition of endothelial integrins and effectively prevents cancer

vascular abnormalities, thus improving the penetration of anti-neoplastic drugs and impairing metastases. In colla-

boration with the Laboratories of Transgenic Mouse Models and Cancer Cell Biology of our Institute, we generated

a parenterally-deliverable mutant Sema3A* protein that, thanks to its ability to bind with high affinity the signaling

receptor plexin A4, is much more active than its wild type counterpart both on cultured ECs and in mouse models

of pancreatic cancer. More recently, we identified: i) a novel Sema that is endowed with an unpredictable in vivo

pro-angiogenic activity ii) a novel signaling pathway that, by coordinating the endocytosis of active integrins and

the exocytosis of freshly synthesized ECM, controls blood vessel formation and patterning both in vitro and in vivo.


Our data provide evidence of how the negative pharmacological modulation of endothelial integrins by Sema3A

can be therapeutically exploited to improve the biodistribution of anti-neoplastic drugs and counteract the hypoxia-

driven metastatic dissemination of cancer cells. It will therefore be crucial to further characterize the mechanisms

and molecular determinants responsible for: (i) the inhibition of integrin-dependent EC adhesion by Sema3A; (ii)

the selective Nrp1-dependent control of active integrin traffic in ECs.

Selected references


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Guido Serini MD, PhD Laboratory of Cell Adhesion Dynamics

Assistant ProfessorUniversity of Torino Medical School


Fax. +39.011.9933524

Serini G, Tamagnone L. Bad vessels beware! Semaphorins will sort you out! EMBO Mol Med. 2015, 7:1251-3.

Valdembri D, Serini G. Regulation of adhesion site dynamics by integrin traffic. Curr Opin Cell Biol. 2012, 24:582-91.

Sandri C, Caccavari F, Valdembri D, Camillo C, Veltel S, Santambrogio M, Lanzetti L, Bussolino F, Ivaska J, Serini G. The R-Ras/RIN2/Rab5 complex controls endothelial cell adhesion and morphogenesis via active integrin endocytosis and Rac signaling. Cell Res. 2012, 22:1479-501.


Over the last two decades, Guido Serini has investigated the role of integrin-mediated cell adhesion in the

control of tumor progression and angiogenesis. He firstly reported (J. Natl. Cancer Inst. 1996) how the neo-

expression of alpha6-beta4 integrin, now well-known for its pro-metastatic activity, as a hallmark of the tran-

sition of human benign adenomas into malignant carcinomas. He also demonstrated (J. Cell Biol. 1998) how

the generation of pro-invasive and pro-angiogenic cancer associated fibroblasts requires integrin-triggered si-

gnals. He then focused on the mechanisms that, by tuning the conformational activation of integrins, control

physiological angiogenesis and are disrupted in cancer, finally resulting in hypoxia-driven metastatization. After

reporting that autocrine VEGF is critical for integrin activation and vascular network formation (EMBO J., 2003),

he showed for the first time how endothelial class 3 semaphorins (SEMA3) signal through plexin receptors to

inhibit integrins and allow vascular morphogenesis (Nature 2003). Next, together with Dr. Giraudo, he discove-

red that SEMA3A is present in endothelial cells of pre-malignant lesions, is lost during tumor progression, and,

when therapeutically reintroduced, is able to normalize the vasculature, to inhibit tumor growth, and to extend

survival (J. Clin. Invest., 2009). He recently unveiled how vascular morphogenesis relies not only on integrin

activation, but also on dedicated signaling pathways that control active integrin traffic back and forth from

adhesion sites (PloS Biol. 2009; Cell Res. 2012; Curr. Opin. Cell Biol. 2012)


New targets for anti-angiogenic therapyThe better understanding of the anti-metastatic and normalizing effects of Sema3s on tumor blood vessels, will allow to design new strategies in order to overcome the resistance to anti-angiogenic therapy

Team Federica Maione, Yaqu Qiu, Marco Soster, Alessia Visentin

Research topic:

Semaphorins: new tools to “normalize” the tumor microenvironment and to halt metastasis formation.

Background:

Angiogenesis is required for invasive tumor growth and metastasis. It is well described that tumor vessel normalization

represents a remarkably advantageous anti-cancer strategy, reducing tumor hypoxia and also being able to favor che-

motherapy delivery and response to radiotherapy. It is critical therefore to identify new “pro-normalizing” modulators to

define new anti-angiogenic combinatory regimens to block tumor growth. In these years, several studies have showed

that class3 semaphorins (Sema3s) - that act via receptor complexes binding neuropilins 1 and 2 (Nrp1/2) and transdu-

cing the signal by plexins (Plxns) - represent new targets to inhibit tumor angiogenesis and cancer growth.


Sema3A has been uncovered as a new vessel normalizing and anti-metastatic agent in mouse models of spontaneous

pancreatic neuroendocrine tumors (RIP-Tag2) of cervical carcinomas (HPV16/E2), of pancreatic adenocarcinoma

(PDAC) and of breast cancer (4T1). Interestingly, the treatment of mice with adeno-associate virus AAV-8-Sema3A

counteracted the resistance to the anti-angiogenic therapies by normalizing the tumor vasculature, inhibiting hypoxia

and several hypoxia-induced pro-metastatic signaling pathways. Sema3A was able to recruit into tumors a sub-popula-

tion of CD11b+, Nrp-1+, GR-1- and Tie-2- monocyte (NEMs) inhibiting tumor progression and to normalize the tumor

vasculature. Notably, Sema3A acted directly on tumor cells. In fact, inhibited HGF-induced Met phosphorylation and

impaired the chemo-invasion of several met-addicted tumor cell lines, by interfering with Met recycling and interna-

lization. Remarkably, Met phosphorylation was strongly inhibited both in vessels and cancer cells in Sema3A-treated

PDAC mouse model. In collaboration with the Laboratories of Cell Adhesion Dynamics of our Institute, we generated a

furin-resistant mutant Sema3A* protein that, binding with high affinity with plexin A4, efficiently inhibited metastasis

formation and normalized the tumor vasculature in mouse models of pancreatic cancer. We uncovered together a novel

Sema that is endowed with an unpredictable in vivo pro-angiogenic activity.


Our findings provide evidences that Sema3A, by acting on several tumor cell types and activating the immune-system,

blocks tumor growth and metastasis spreading in different tumor types. Based on these findings the main focus of

research are the following: (i) investigation of the molecular mechanisms by which Sema3A turns off the HGF/Met

pathway in different stroma cell types and cancer cells; (ii) evaluation of new therapeutic combinatorial strategies to

enhance with Sema3A (and other Semas) the anti-tumor immune response in several mouse models of cancers; (iii)

evaluation of the anti-metastatic and pro-vessel normalizing effects of mutant Sema3A* protein in different human

mouse models of pancreatic, breast, cervical, and colon cancer.

Selected references


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Enrico Giraudo PhDLaboratory of Transgenic Mouse Models

Assistant Professor University of Torino Medical School


Fax. +39.011.9933524

Maione F.,et al. The cholesterol biosynthesis enzyme oxidosqualene cyclase is a new target to impair tumour angiogenesis and metastasis dissemination. Sci. Rep. 2015, 5,9054.

Serini G.,et al. Class 3 semaphorins: physiological vascular normalizing agents for anti-cancer therapy. J. Intern. Med. 2013, 273:138-55.

Maione F.,et al. Semaphorin 3A overcomes cancer hypoxia and metastatic dissemination induced by antiangiogenic treatment in mice. J. Clin. Invest. 2012, 122:1832-48.


Enrico Giraudo has contributed with a high quality track record of publications to the research field of tumor

angiogenesis. Since the beginning, he has contributed to the identification of the molecular mechanisms that

regulate the motility and differentiation of angiogenic endothelial cells (J. Biol. Chem, 1998; Nature Med. 1996;

J Biol Chem. 2003). His main focus has been the study of the mechanisms regulating tumor angiogenesis and

cancer progression in transgenic mouse models of tumorigenesis. He described the molecular zip-code de-

ciphering blood and lymphatic heterogeneity during tumorigenesis (Cancer Cell. 2003). Then, he uncovered an

important role of matrix metallo-proteases (MMP)-9 and macrophages on tumor angiogenesis in a transgenic

model of cervical cancer, by employing an amino-bisphosphonate Zoledronic acid able to efficiently inhibit can-

cer growth (J Clin Invest. 2004). More recently he demonstrated that the axon guidance cue Semaphorin 3A is

an endogenous angiogenic inhibitor and that its therapeutically re-expression in cancers impairs angiogenesis,

tumor growth and induced vessel normalization in mouse models of cancer (J Clin. Invest. 2009). On these

bases, he showed that Sema3A was able to overcome resistance to anti-angiogenic therapies, by normalizing

tumor vessels and inhibiting hypoxia (J. Clin. Invest. 2012).


Role of membrane receptor endocytosisin the dissemination of metastatic cellsKnowing the mechanisms that control the downregulation of receptors involved in metastatic cell dissemination provides the rationale for the generation of innovative anti -neoplastic drugs

Team Daniele Avanzato, Emanuela Pupo, Nadia Ducano

Research topic:

Role of endocytosis in the regulation of signaling by oncogenic tyrosine kinase receptors.

Background:

The increased activity of receptor tyrosine kinases (RTKs) is recognized as one of the most common alteration

in cancer. This is mainly achieved by the occurrence of activating mutations, or by gene amplification. Beside

these mutational events, other mechanisms may occour to increase the RTKs activity. Among them endocytosis

and receptor degradation play a relevant function. Although mutations in endocytic genes have crucial roles in

the manifestation of the neoplastic phenotype, the possibility to exploit endocytic proteins as targets of phar-

macological intervention, alone or in combination with other treatments, are still poorly investigated. Research

in our unit addresses the role of key endocytic players, the small GTPase Rab5 and its negative regulator RN-tre,

as well as the effects of endocytosis inhibition, in cancer cell proliferation and invasion.


We found that withdrawal of tyrosine kinase inhibitors can exert rebound cancer cell proliferation and in vivo

tumor growth. Cancer cells bearing the amplification of the MET receptor depend on MET for growth and sur-

vival. Treatment with a specific kinase inhibitor halts their proliferation, but, upon inhibitor discontinuation,

MET phosphorylation and downstream activation resume with higher levels if compared to untreated cells. This

generates a “rebound” effect pushing quiescent cancer cells back into the cell cycle both in vitro and experi-

mental tumor models in vivo. We investigated the function of RN-tre in breast cancer cells. We found that RN-tre

silencing reduces AKT phosphorylation, while its overexpression in a normal mammary cell line promotes 3D

overgrowth of acinar-like sphere.


Database analyses has revealed that RN-tre is amplified/overexpressed in 16% of breast tumors. Based on its

function in receptor signaling and cell adhesion, we hypothesize that RN-tre overexpression in breast cancer

cells might confer proliferative advantages by upregulating oncogenic tyrosine kinase receptors, and/or promo-

ting anchorage-independent growth and cell survival.

Based on our findings on MET kinase inhibition, we plan to address the effects of derailed endocytosis in tumor

recurrence. We will study the rebound effects on proliferation of cancer cells that were initially subjected to ATP-

competitive RTK inhibitors and then released from the inhibition, trying to mimic what might happen in patients

treated with similar drugs in a time window. We will address whether alternate or metronomic treatment with

RTKs inhibitors and therapeutic antibodies might counteract this harmful effect in vivo.

Selected references


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Letizia Lanzetti PhDLaboratory of Membrane Trafficking

Assistant Professor University of Torino Medical School

E-mail: [email protected]. +39.011.9933508

Fax. +39.011.9933524

Serio G, et al. Small GTPase Rab5 participates in chromosome congression and regulates localization of the centromere-associated protein CENP-F to kinetochores. Proc Natl Acad Sci U S A. 2011; 108:17337-42.

Palamidessi A, et al. The GTPase-activating protein RN-tre controls focal adhesion turnover and cell migration.Curr Biol. 2013 ;23:2355-64

Rolland Y.,et al. The CDC42-interacting protein 4 controls epithelial cell cohesion and tumor dissemination.Dev Cell. 2014 ;30:553-68.


Letizia Lanzetti completed her training at the Department of Medical Genetic in Torino where she participated

in the isolation of a novel gene family: the endophilins. Next, she joined the European Institute of Oncology in

Milan to study the impact of endocytosis on the signal transduction cascade. Her work pioneered the existence

of crosstalk between these processes as described in two of her major publications (Nature, 2000; Nature,

2004). In 2008 she established her laboratory at the Candiolo Cancer Institute. Here, she revealed a novel fun-

ction for endocytic proteins in cancer cell division (PNAS 2011), and in cell migration and invasion (Curr. Biol.

2013).

Her work has led to the identification of molecular pathways that control signaling and trafficking of receptors

involved in cell transformation and invasion. Currently, she is widening her research looking at the feasibility of

targeting endocytic molecules, in combination with other treatments, in cancer therapy.

RN-tre overexpression causes overproliferation in 3D matrices

DAPI + caspase3 caspase 3 HA

CTR

RN-tre


From neuronal biology to neoplastic progressionThe identification of ‘molecular keys’ that mediate the relationship between tumor cells, blood vessels and nerves will contribute to the definition of a new class of therapeutic targets

Team Laura Bizzozero, Margherita Pergolizzi, Elena Riccitelli

Research topic:

Neuroligin: a novel modulator of cell transformation and cancer diffusion through nerves.

Background:

Neuroligin (NLGN) is a neuronal and tumoral adhesion protein. Tumor-nerve interactions are a clinically signi-

ficant, but often underestimated way of cancer diffusion, including colorectal cancer (CRC). Tumor and nerve

relations occour through Perineural Invasion (PNI), namely the invasion of nearby nerves by cancer cells, and

neo-neurogenesis, or the stimulation of neurite outgrowth by cancer through soluble signals. These aspects

represent different conditions of a wide range of molecular interactions that are nevertheless poorly defined.

There is no specific therapy able to target tumor nerve interactions. Therefore, new molecular players and

therapies are needed.


This laboratory has been working on the extra-neuronal activities of NLGN for the last ten years. Since 2013

we have focused on the following questions on the role of tumoral/nervous NLGN (mainly in CRC): (a) How

does it impact tumor “autonomous” cell behaviour? (b) Does it modulate PNI and neo-neurogenesis? The

results obtained to date show that NLGN inhibits tumor cell proliferation, anchorage-independent growth and

in vivo tumor growth but, conversely, promotes PNI.


If broadly confirmed, our data reveal NLGN as a double-faced cue (i.e. growth suppressing but pro- invasive).

We now want to exploit the knowledge coming from the neuronal field, the large amount of reagents accu-

mulated in this laboratory, and both in vitro and in vivo experimental settings (mouse xeno- and ortho- tran-

splants) in order to fully answer the questions stated above.

Selected references


Can

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Marco Arese PhDLaboratory of Neurovascular BiologyAssociate Professor of BiochemistryUniversity of Torino Medical School


Fax. +39.011.9933524

Samarelli AV, et all. Neuroligin 1 induces blood vessel maturation by cooperating with the α6 integrin. The Journal of biological chemistry. 2014, 289:19466-76

Rissone A, et al. The synaptic proteins neurexin and neuroligin synergize with extracellular matrix-binding vascular endothelial growth factor a during zebrafish vascular development. Arterioscler Thromb Vasc. Biol. 2012, 32:1563-72. Bottos A, Rissone A, Bussolino F, Arese M. Neurexins and neuroligins: synapses look out of the nervous system. Cell Mol Life Sci. 2011, 68:2655-66.


Marco Arese has dedicated his early research activity to the study of various mediators of intercellular communi-

cation that involve cancer and vascular cells (J Clin Invest. 1995). In the laboratory of Prof. Daniel B. Rifkin, at the

New York University Medical Center (USA), he examined the mechanisms of action of two growth factors: Fibro-

blast Growth Factor-2 (Mol Biol Cell. 1999) and transforming growth factor-beta (TGF-beta). For the last fifteen

years, he has devoted himself to studying the “molecular parallels” between the nervous system and vascular and

tumoral tissues, with the aim of finding new targets that drive the cooperation between the tumor and its “micro-

environment” (specifically, infiltrating vessels and nerves). In turn, this cooperation is known to fuel disease pro-

gression. Initially, he discovered that two synaptic proteins, Neurexin and Neuroligin, have widespread functions

in the vascular system, including tumor angiogenesis (Proc Natl Acad Sci U S A. 2009; Arterioscler Thromb Vasc

Biol, 2012). He is currently focused on the role these two nervous proteins - still widely underestimated - play in

the tumor/nerves connections.


Cellular and molecular mechanisms sustaining tumor angiogenesisAll target therapies, including antiangiogenic therapies, are far from achieving the wished results. However, antiangiogenic regimens are an important tool for solid tumor treatment. Our studies are aimed at understanding the complexity of angiogenic processes in physiology

TeamSerena Marchiò, Elena Astanina, Valentina Comunanza, Gabriella Doronzo, Lucia Napione, Alessio Noghero, Davide Corà, Maria Alvaro, Anna Gualandris, Stefania Rosano, Emanuele Middonti, Emanuela Doronzo

Research topic:

Identification of early biomarkers of pancreatic ductal carcinoma (PDAC).

Background:

Elegant tracking studies of the genetic evolution of PDACs indicate at least 15 years between the occurrence of

the initiating mutation and the acquisition of metastatic ability. This result demonstrates a broad time window

of opportunity for early detection to prevent deaths from metastatic disease. We reasoned that an early bio-

marker had to play a necessary role in the onset of the disease and it had to be preferentially released from the

cells. Therefore we performed a whole transcriptomic analysis of the early in situ carcinomas (PanIN) isolated

by laser capture microdissection from a genetically engineered mouse model characterized by the expression

of KRASG12V in acinar cells through the use of the elastase promoter (Elas-K-RasG12V).


RNAs highly expressed in PanIN encode proteins involved in axon guidance cues and synaptogenesis that

have been recently demonstrated to be altered in PDAC. We focused on neuroligins (Nlg), a family of adhesive

molecules regulating synaptic activity. Anti-pan-Nlgs antibody showed a faint membrane signal of acinar and

ductal cells whereas PanIN cells displayed a stronger staining that dropped in PDAC. Similar results were obtai-

ned in human lesions. By analysing the expression of the 5 Nlgs we visualized a minute population of acinar

cells carrying KRasG12V and expressing Nlg-2 associated with stemness markers (DCLK1, CD24, CD44, and

CXCR4). The current hypothesis of the pathogenesis of PDAC relies on the differentiation of acinar precursors

carrying KRAS mutation into ductal cells (acinar-ductal metaplasia). We demonstrated that KRasG12V acinar

cells isolated from Elas-KRasG12V underwent in vitro acinar ductal metaplasia with an increased expression

of Nlg-2. An isogenic cell line carrying KRasG12V overexpressed Nlg-2 if compared with wild-type cells. Loss-

of-function experiments showed that Nlg-2 ablation halted acinar-ductal metaplasia, supporting the idea that

Nlg-2 is necessary in the early phase of PDAC. Then, we considered the possibility that Nlg-2 was shed by the

cells, supported by the presence of a Disintegrin And Metalloproteinase (ADAM) proteolytic cleavage site in

the extracellular domain. Nlg-2 accumulated in the serum-free supernatant of pancreatic PT45 human cell line

overexpressing Nlg-2. This phenomenon is blocked by ADAM inhibitors TAPE-2 and GM 6001.


We reported that Nlg2 (i) has an early role in PDAC by promoting the acinar-ductal metaplasia of precursors; (ii) is

shed by an ADAM-dependent mechanism. To further support the role of Nlg2 as an early biomarker in PDAC, we will

continue the following activities: (i) demonstration of the role of Nlg-2 in stemness features of acinar precursors

undergoing acinar-ductal metaplasia; (ii) analysis of PDAC progression in Elas-K-RasG12V model crossed-back with

Nlg-2-/- mice (iii) generation of mAb anti-Nlg2 to set-up an ELISA for Nlg-2 quantification in biological fluids; (iv)

increase of human samples to analyse Nlg-2 expression and activities.

Selected references


Can

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stitute

Federico Bussolino MDLaboratory of Vascular Oncology

Full Professor of BiochemistryUniversity of Torino Medical School


Fax. +39.011.9933524

Bussolino F, Giraudo E, Serini G. Class 3 semaphorin in angiogenesis and lymphangiogenesis. Chem Immunol Allergy. 2014, 99:71-88

Corà D, Astanina E, Giraudo E, Bussolino F. Semaphorins in cardiovascular medicine. Trends Mol Med. 2014, 20:589-98.

Marchiò S, Astanina E, Bussolino F. Emerging lymphae for the fountain of life. EMBO J. 2013,32:609-11

Napione L et al. IL-12-dependent innate immunity arrests endothelial cells in G0-G1 phase by a p21(Cip1/Waf1)mediated mechanism. Angiogenesis. 2012, 15:713-25


Federico Bussolino began his scientific career by investigating the cross-talks between the mediators of inflamma-

tion and vascular cells with the goal of understanding the cellular traffic across vascular walls. Then (1988), he was

captured by the discovery of colony stimulating factors and their strict specificity during hematopoiesis. He has gone

beyond this concept, showing that these molecules activate a pro-survival and pro-angiogenic program in endothelial

cells (Nature, 1989). After 20 years, the proangiogenic roles of these molecules have clarified their fundamental

role in the bone marrow vascular niche, which is fundamental in the maturation of blood cells. Then he moved to

tumor angiogenesis showing the role of the MET receptor in cancer-associated vascularization (J Cell Biol, 1992).

This seminal observation supported the concept that MET is pivotal in regulating cancer invasive growth. In 1996,

he began to work on VEGFs showing that HIV-1 Tat activates the VEGF pathway. This result emphasizes the ability of

the virus to directly support the progression of tumors appearing in AIDS (Nat Med., 1996; Nat Med, 2002). He fir-

stly demonstrated (Embo J.,1999) the existence of a dynamic platform constituted by adhesive and tyrosine-kinase

receptors. In 2003, he funded a successful interdisciplinary PhD program on complexity in biology that has antici-

pated the concepts of Systems Biology. In 2003, he contributed to open a frontier in angiogenesis, by connecting

the 3D architecture of the vascular bed with the mechanisms that wire neurons. (Nature, 2003). Recently, he has

added another piece of information about the cues shared by the brain and vascular system by demonstrating that

two synaptic proteins, neurexin and neuroligin, regulate the assembly of vascular units.


Cell migration in tumor angiogenesis and invasionThe identification of molecular mechanisms involved in tumor cell migration will allow the prevention of metastatic dissemination by pharmacological inhibition

Research topic:

Molecular mechanisms that differentiate invasive cancer cells from their non-motile counterparts, and the rela-

tionship between tumor and non-tumor cells during cell migration.

Background:

Cancer cells spread from the initial site of tumor growth acquiring an invasive phenotype characterized by both

the loss of cell-cell interactions and increased cell motility. Cells undergoing this epithelial to mesenchymal

transition are then able to move and spread throughout the entire body as isolated and highly motile cells. Re-

cent evidence shows that tumor cells also move as groups both in normal development and in cancer models,

in a process named “collective migration”. Even vascular endothelial cells collectively migrate into tumor mass

forming new vessels in a process called “tumor angiogenesis”.


We are developing new three-dimensional (3D) culture systems that in combination with conventional cell bio-

logy approaches, allow the study of collective and directional cell migration. A new model for in vitro tumor

angiogenesis studies has been described and cell spheroid cultures from breast and prostate cell lines have

been developed. With these three-dimensional models we have been able to discover new mechanisms of PI3K

regulation of collective migration and the role of the endothelial podosome in tumor angiogenesis.


We believe that spheroids/organoids 3D culture from cells and tissues could overcome limitations of established

cell lines, such as adaptation to 2D growth, providing better models to study collective migration, epithelial-to

mesenchymal transition and tumor vascularization, and the identification of molecular mechanisms involved in

these processes. Moreover, we are establishing 3D cell cultures from patient-derived xenograft (PDX) of colon

cancer with a high percentage of in vitro engraftment. By combining our know-how in 3D cultures with the PDX

platform developed in the Institute, we will implement an unprecedented in vitro platform for genetic functional

and drug screening purposes.

Team Laura Di Blasio, Alberto Puliafito, Giulia Chiaverina, Paolo Gagliardi, Desiana Somale

Selected references


Can

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Luca Primo PhDLaboratory of Cell Migration



Fax. +39.011.9933524

di Blasio L.,et al. PDK1 regulates focal adhesion disassembly by modulating endocytosis of avß3 integrin. J Cell Sci. 2015,128:863-77

Puliafito A., et al. Three-dimensional chemotaxis-driven aggregation of tumor cells. Sci Rep. 2015, 5:15205

Gagliardi PA, di Blasio L, Primo L. PDK1: A signaling hub for cell migration and tumor invasion. Biochim Biophys Acta. 2015, 1856:178-88


Luca Primo has a remarkable experience in the field of vascular cell biology. He started his research experience

characterizing the thrombospondin receptor CD36. He showed that this membrane protein was involved in the

pathogenesis of cerebral malaria (Blood, 1993; J Immunol, 2001) and angiogenesis (Faseb J., 2005) During

his post-doc he contributed to unveil the molecular mechanisms of several angiogenic growth factors, such as

VEGF-A (JBiolChem, 1998) and GM-CSF (Blood, 1997). He then moved to the Candiolo Cancer Institute where

he focused his research efforts on the PI3K signalling pathway in angiogenesis and cell motility (Oncogene,

2000). During a research experience at the Tuft University of Boston, he started to study the protein kinase

PDK1, a downstream effector of PI3K. He discovered the central role of PDK1 in cell directional motility (JCell-

Biol, 2007) and he elucidated the function PDK1 in breast cancer growth (Neoplasia, 2012) and invasion (JCell-

Biol, 2014). By gene expression analysis in endothelial cells he discovered the pro-angiogenic role of integrin

α6 (Cancer Res, 2010) and the effects of miR126 on Angiopoietin1 signaling (BBA-MolCellRes, 2012). Moreo-

ver, he developed a new model for the three-dimensional study of tumor angiogenesis, the Human Arterial Ring

assay, which can be exploited for drug screening and gene function analysis on human vessels (Blood, 2013).

More recently, he described regulatory mechanisms of integrin endocytosis (Traffic, 2010; JCellSci, 2015) and

the role of endothelial podosomes in angiogenesis (NatCellBiol, 2014). Luca Primo is Associate Professor of

Biochemistry and group leader of the Laboratory of Cell Migration.

Different hallmarks of tumor invasion are controlled by PDK1

BM BM

InvadopodiaFormation

PDK1 REGULATES TUMOR CELLINVASION BY PROMOTING

Collectiveinvasion

Amoeboid invasion


Exploiting glucose ‘addiction’ of KRAS-mutant colorectal tumors for cancer therapyThe development of novel approaches such as anti-metabolic therapy can open new paths to the treatment of aggressive forms of colorectal cancer

Team Manuela Cazzanti, Alessia Mira, Virginia Morello

Research topic

Enhanced glucose uptake and increased glycolysis are hallmarks of tumor cells. Building on key preliminary

results generated in the past years, the Laboratory of Cancer Metabolism is studying a strategy for exploiting

these peculiar metabolic characteristics for cancer therapy.

Background

KRAS-mutant colorectal tumors display an exceptionally high glycolysis rate, which makes them ‘addicted’ to

glucose. This dependency suggests a potential therapeutic approach based on anti-glycolytic agents, including

drugs used for treating diabetes. However, preliminary experiments conducted in our lab suggest that Hepa-

tocyte Growth Factor (HGF), a cytokine that is abundant in the tumor micro-environment, can rescue KRAS-

mutant CRC cells from starvation-induced apoptosis. Based on this evidence, we hypothesized that inhibition of

HGF or its receptor MET would render KRAS-mutant tumors more sensitive to anti-metabolic therapy.

Achievements

We selected a panel of metabolism-targeting drugs that affected glucose metabolism by different mechanisms

and at different levels, including metformin and phenformin (both inhibitors of mitochondrial complex I), dich-

loroacetate (an inhibitor of pyruvate dehydrogenase kinase), and WZB-117 (an inhibitor of the facilitative gluco-

se transporter GLUT1). Pharmacological analysis of these drugs in isogenic KRAS CRC cell lines revealed that

CRC cell clones expressing mutant KRAS are dramatically more sensitive than their wild-type KRAS counterpart

to glucose deprivation or to treatment with metformin, phenformin, dichloroacetate or WZB-117. Apoptosis

analysis showed that all of these anti-metabolic agents were causing a pronounced increase in the percentage

of apoptotic cells, which could be prevented or mitigated by recombinant HGF. In an orthotopic mouse model

of KRAS-mutant CRC using human HGF knock-in SCID mice, metformin inhibited tumor growth and inhibited

hepatic and pulmonary metastasis. Moreover, anti-metabolic drugs cooperated with the HGF-neutralizing anti-

body ficlatuzumab in inhibiting tumor growth and in suppressing metastasis.

Conclusions and perspectives

These preliminary data strongly suggest that KRAS-mutant CRC cell metabolic vulnerability can be exploited

therapeutically by interfering with glucose metabolism. They also point at micro-environment-derived HGF as a

major source of resistance to any kind of therapy that causes metabolic stress to KRAS-mutant CRC cells. Re-

search in the Laboratory of Cancer Metabolism focuses on how to transfer this knowledge into clinical practice.

Selected references


Can

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stitute

Paolo Michieli PhDLaboratory of Cancer Metabolism



Fax. +39.011.9933225

Hultberg A, et al., Depleting MET-Expressing Tumor Cells by ADCC Provides a Therapeutic Advantage over Inhibiting HGF/MET Signaling. Cancer Res. 2015; 75:3373-3383

Pennacchietti S, et al.,Microenvironment-derived HGF overcomes genetically determined sensitivity to anti-MET drugs. Cancer Res. 2014; 74:6598-6609.

Basilico C, et al., Four individually druggable Met hotspots mediate HGF-driven tumor progression. J Clin Invest. 2014; 124:3172-86.


Paolo Michieli graduated in Biological Sciences at the University of Milano in 1990. After a few years of training

at the Mario Negri Institute for Pharmacological Research and the Istituto Nazionale dei Tumori, both in Milano,

he carried out his post-doc in the Laboratory of Cellular and Molecular Biology at the National Cancer Institute

in Bethesda, Maryland. During this period he contributed to several scientific publications in the field of on-

cogenes, growth factors and cell cycle. In 1997 he moved back to Italy to the newly founded Candiolo Cancer

Institute in Candiolo, Torino. Since then he has been Group Leader of a small team committed to experimenting

novel therapeutic strategies in oncology, with particular reference to the molecular targeting of growth factors

and their receptors. In 2001 he specialized in Clinical Pathology at the University of Torino Medical School.

In 2008 he received a Ph.D. in Cellular Sciences and Technologies from the same University, and in December

2011 he was appointed tenured Assistant Professor at the Department of Oncology.

30 | From Molecular Biology to “Precision Medicine”

Cancer Stem Cell Laboratory 2015

From Molecular Biology to “Precision Medicine” | 31

Research topic 2From Molecular Biology to “Precision Medicine”

Research topic 2 is the core scientific basis for most of the Institute’s research projects.

Advances in genome sequencing and expression analysis have made it possible to quickly identify genetic alterations with a potential “driver” role in tumor development, with unprecedented informative power. Thanks to these studies, today it is possible to classify tumors, not only according to their site of origin and histopathological features, but also – and especially – according to the identification of the genetic lesions supporting their growth. Genetic lesions’ identification is crucial to kill tumors by using targeted therapies. However, the contribution of each lesion to the transformed phenotype remains elusive. Moreover, there is still insufficient knowledge of the mechanisms that control the lack of response to targeted therapies, even in the presence of a molecular target (primary resistance), as well as of the mechanisms that lead to progressive attenuation of the response after prolonged treatment (acquired resistance). Translational Research is a multi-disciplinary task based on the expertise of a new generation of scientists, specifically trained to translate molecular information into more effective treatments. The goals are ambitious: to integrate the traditional prognostic and diagnostic factors with a very detailed characterization of the genetic and functional alterations of the tumor; to identify new malfunctioning regulatory pathways in cancer; to isolate and study cancer stem cells; to develop new preclinical platforms that can reliably disclose – and understand in detail – the prospective results of clinical practice. The generation of this kind of knowledge is instrumental to instructing clinical trials that will be no longer based on empirical information but on a strong rational connotation.

Research Topic 2 concerns:2.1. Oncogenes and growth factors2.2. Genes responsible for response to antineoplastic drugs2.3. Preclinical models of “personalized” therapies2.4. Resistance to targeted therapies


Oncogenes involved in invasive growthGenetics and molecular biology underlying the metastatic process are largely unknown. It is mandatory to identify and to validate therapeutic targets to develop long-sought (and still missing) effective therapies

TeamAnna Cristina Basilico, Silvia Benvenuti, Alessandra Gentile, Federica Verginelli, Maria Rita Virzì, Chiara Modica, Melissa Milan

Research topic:

Oncogenes involved in invasive growth

Background:

Growing evidence prompted us to hypothesize that the metastatic process involves members of the gene

‘super-family’ of tyrosine kinase receptors for ‘scatter factor’, homologues to the Met oncogene (Ron, Axl, Mer

and Tyro). Recently, other poorly characterized genes of the family have been identified, Ror1 and Ror2, that can

possibly interfere with the response to drugs directed against Met (entered in the clinic), generating resistance.


In the past we have shown that the Met oncogene encodes the receptor for the ‘scatter factor’ HGF. A series of

studies - performed for two decades in this and other laboratories - revealed the involvement and the importan-

ce of this oncogene in human cancers, demonstrating its key role in the control of ‘invasive growth’, a genetic

program driving the metastatic process. We contributed to the development of drugs that inactivate the Met

tyrosine kinase: either chemical inhibitors or monoclonal antibodies. These drugs inactivate cancer stem cells

in preclinical models of glioblastoma, colorectal and gastric carcinomas, and provide promising results in cli-

nical trials.


We are now working on the characterization of tyrosine kinase receptors of the Met oncogene family (Ron, Axl,

Mer and Tyro, Ror1-2) at genetic, biochemical and functional levels. In particular, we are studying their possi-

ble interference - positive or negative - with the intracellular signaling triggered by Met during execution of the

invasive growth program. Emphasis is given to the study of a hyper-metastatic syndrome, the so-called CUP,

metastatic cancer from unknown primary origin.

Selected references


Can

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Paolo Comoglio MDLaboratory of Molecular Therapy Exploratory Research

Full ProfessorUniversity of Torino Medical SchoolE-mail: [email protected]

Phone. +39.011.9933601 Fax. +39.011.962.1525

Boccaccio C, Comoglio PM. MET, a driver of invasive growth and cancer clonal evolution under therapeutic pressure. Curr Opin Cell Biol. 2014 31:98-105.

Boccaccio C, Luraghi P, Comoglio PM. MET-mediated resistance to EGFR inhibitors: an old liaison rooted in colorectal cancer stem cells. Cancer Research. 2014. 74:3647-51

Benvenuti S, Gentile A, Lazzari L, Arnesano A, Trusolino L, Comoglio PM. An ‘in-cell trial’ to assess the efficacy of a monovalent anti-MET antibody as monotherapy and in association with standard cytotoxics. Mol Oncol. 2014, 8:378-88

Biographical Sketch

Paolo Comoglio has a long and distinguished record in the field of research on tyrosine kinase receptors and

related oncogenes. He developed the first anti-phosphotyrosine antibody (EMBO J., 1984); through this tool he

identified the tyrosine kinase encoded by the rearranged Bcr-Abl oncogene, responsible for the onset of Chronic

Myeloid Leukemia (Mol Cell Biol. 1986)

He focused his research efforts on the full mechanistic insight of the genetics, biochemistry and biology of

the Met receptor, with a special emphasis on the role of this oncogene in human cancers. After a long standing

career he is now acknowledged as a leader in the field. He identified the tyrosine kinase receptor encoded by

the Met oncogene (Nature, 1989), discovered that Hepatocyte Growth Factor (HGF) is the cognate ligand (On-

cogene, 1991) and cloned the Plexin genes (Cell, 1999), recognizing their role as receptors for Semaphorins,

signals for cell-cell repelling clues. He identified and elucidated the functions of two tyrosine kinase receptors,

encoded by genes structurally related to Met: Ron, (EMBO J, 1994), and an homologous kinase-dead receptors,

Ror ( Cancer Res, 2011).

On the translational front he generated the anti-Met antibody DN30, and its monovalent counterpart, to be used

in cancer therapy, either conventional or by ‘gene transfer’ (Cancer Res, 2008).

Paolo Comoglio expounded and introduced a number of insights that are now largely accepted and widespread,

notably the concept of ‘invasive growth’, a genetic program otherwise ‘physiological’ but ‘usurped’ by cancer

cells to progress toward metastasis (Nature Rev. Cancer, 2002)


Preclinical models of oncogene ‘addiction’We aim to identify ‘dominant’ genetic aberrations that drive tumorigenesis and put them in the context of concurrent ‘recessive’ alterations that may act as modifiers of response to targeted therapies

Team Andrea Bertotti, Barbara Lupo, Francesco Galimi, Francesco Sassi, Giorgia Migliardi, Francesca Cottino, Simonetta Maria Leto, Eugenia Zanella

Research topic:

Translational models of cancer precision medicine.

Background:

Advances in technologies have allowed the attainment of powerful insights into the molecular determinants of

human tumours, and in many cases inactivation of individual oncogenic drivers results in tumor regression.

However, this knowledge has been translated into effective treatments very slowly, due to difficulty in predicting

how the complex mutational background and the adaptive resilience of cancer cells can influence the activity

of the dominant oncogene and modify response to therapies.


Our projects aim at exploring the mechanisms of tumor dependency on oncogenic drivers and at understan-

ding how such dependency is affected by genomic or functional modifiers, with an emphasis on colorectal

cancer (CRC). Our experimental pipeline entails the deployment of multi-dimensional data for discovery and

hypothesis validation, followed by cell-based mechanistic investigation and preclinical validation in patient-

derived tumorgrafts – ’xenopatients’. By this approach, we have contributed to finding that hyperactivation of

the HER2 and MET oncogenes correlate with resistance to anti-EGFR therapies (cetuximab and panitumumab).

To examine the global effects of somatic genetic changes in CRC on sensitivity to EGFR-targeted therapies, we

performed complete exome sequence and copy number analyses of 129 tumors and analyzed their response

to anti-EGFR blockade in xenopatients. We identified several alterations as potential mechanisms of resistance

to this therapy as well as genetic lesions segregating in tumors with increased sensitivity to anti-EGFR therapy.

Resistance to EGFR blockade could be overcome in xenopatients through combinatorial therapies targeting

actionable genes. A typical response of CRC patients sensitive to EGFR neutralization is stable disease rather

than massive regression. Mining of candidate gene expression outliers – coupled with retrospective analysis

of patient material and in vitro functional studies – identified IGF2 as a predictor of poor sensitivity to cetuxi-

mab and as a new target. One limitation of xenopatients is that human tumour stroma is substituted by host

(murine) components. However, this drawback can be exploited analytically to dissect the representation of

human (cancer-cell specific) versus mouse (stromal-cell specific) traits in CRCs. By doing so, we contributed to

re-categorising the transcriptional classification of CRC by demonstrating that a subtype displaying features of

epithelial-to-mesenchymal transition and poor prognosis is not an expression of aggressive cancer cell dediffe-

rentiation (as originally argued) but rather a read-out of abundant stromal content.


Our studies provide a systematic functional approach to evaluate response to targeted therapies in human can-

cer, highlight new mechanisms of responsiveness to anti-EGFR therapies, and provide a new vocabulary for the

molecular management of colorectal cancer with immediate clinical implications.

Selected references


Can

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Livio Trusolino MDLaboratory of Precision Medicine

Associate Professor University of Torino Medical School


Fax. +39.011.9933225

Bertotti A, et al.The genomic landscape of response to EGFR blockade in colorectal cancer. Nature. 2015 8;526:263-7

Zanella ER, et al. IGF2 is an actionable target that identifies a distinct subpopulation of colorectal cancer patients with marginal response to anti-EGFR therapies. Sci Transl Med. 2015 28;7:272ra12.

Trusolino L, Bertotti A. Compensatory pathways in oncogenic kinase signaling and resistance to targeted therapies: six degrees of separation. Cancer Discov. 2012 2:876-80.


Since 2006, Livio Trusolino has been Associate Professor of Histology at the University of Turin Medical School,

Italy. He received his M.D. (1993) from the University of Turin and a Ph.D. in Human Oncology (1997) from the

San Raffaele Scientific Institute, Milan, Italy.

Livio initially studied the cross-talk between growth factor receptors and adhesion molecules in tumour tran-

sformation and cancer progression. Later, his work concentrated on the characterisation of the molecular

mechanisms underlying responsiveness to anti-cancer targeted therapies, using innovative preclinical models.

The results of Livio’s research have been published in leading journals such as Cell, Nature Genetics, Science

Translational Medicine, and Cancer Discovery. He has also written review articles for Nature Reviews Drug Di-

scovery, Nature Reviews Cancer, and Nature Reviews Molecular and Cellular Biology.

Histological section of a colorectal tumor. Nuclei of actively proliferating cells are stained in brown


The puzzling oncosuppressive function of oncogenesIdentification of novel therapeutic targets and predictive biomarkers responsive to chemotherapy or targeted drugs in ovarian cancer

TeamAnnalisa Lorenzato, Martina Olivero, Jessica Erriquez, John David Konda, Erika Torchiaro

Research topic:

Unleashing the intrinsic tumor suppressive properties of oncogenes to kill cancer cells.

Background:

Oncogenic tyrosine kinases (TKs) might harbor intrinsic tumor suppressive functions. They sit at the apex of

multiple downstream signaling pathways that exert various biological effects depending upon cell type and

context. While some of these pathways are mitogenic and pro-survival, others might restrain the oncogenic

potential by promoting, for example, apoptosis and senescence. Obviously, the latter programs do not confer

a selective advantage and thus are hidden in cancer cells. However, intrinsic tumor suppression of TKs might

be unleashed to kill cancer cells. This Laboratory had demonstrated that activation of the MET TK by its ligand

Hepatocyte Growth Factor (HGF) results in the commitment to death of ovarian cancer cells. This model has

been exploited to identify actionable molecules, that could be targeted to treat ovarian cancer and to sensitize

ovarian cancer to standard platinum and taxane based treatment, once they have developed resistance.


Among the actionable molecules this laboratory has identified (i) CDT2, (ii) the small heat shock protein of 27

Kda (HSP27), (iii) the PIM 2 kinase and (iv) the 90 kDa ribosomal S6 kinases (RSKs). In detail, using transcrip-

tomic and small scale proteomics we found that MET activation in ovarian cancer cells resulted in the down

regulation of the anti-apoptotic small heat shock protein of 27 KDa (HSP27, HSPB1) while MET inhibition in

addicted cancer cell lines resulted in the increase of HSP27 and cell protection from death. Likewise, in human

cancer cells susceptible to EGFR inhibition, EGFR inhibitors induced HSP27 expression and were strengthened

by HSP27 suppression. Therefore, in cancer therapies targeting the MET or the EGFR pathways, the induction of

HSP27 might limit the efficacy of targeted agents, alone or in combination with chemotherapy. A SILAC-based

approach allowed the identification of PIM2 as a kinase whose phosphorylation is regulated by DNA dama-

ging agents in ovarian cancer cell lines. Targeting PIM2 kinase sensitized ovarian cancer cells to drug-induced

apoptosis, while PIM2 over-expression resulted in cell resistance to DNA damaging agents. The study of the

RSKs has clarified their role not only in sustaining the metastatic phenotype of ovarian cancer cells but also in

counteracting standard chemotherapy.


Altogether data showed that pro-apoptotic pathways can be unveiled in cancer cells by studying the unexpected

pro-death outcome of MET activation. This allowed the identification of actionable molecules for which inhibi-

tors are available and may find clinical application as an alternative or an adjunct to standard chemotherapy.

Selected references


Can

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ancer In

stitute

Maria Flavia Di Renzo MDLaboratory of Cancer Genetics

Full ProfessorUniversity of Torino Medical SchoolE-mail: [email protected]

Phone. +39.011.9933343 Fax. +39.011.9933524

Olivero M, Dettori D, Arena S, Zecchin D, Lantelme E, Di Renzo MF. The stress phenotype makes cancer cells addicted to CDT2, a substrate receptor of the CRL4 ubiquitin ligase. Oncotarget. 2014, 5:5992-6002. Musiani D, Hammond DE, Cirillo L, Erriquez J, Olivero M, Clague MJ, Di Renzo MF. PIM2 kinase is induced by cisplatin in ovarian cancer cells and limits drug efficacy. J Proteome Res. 2014, 13:4970-82. Musiani D, Konda JD, Pavan S, Torchiaro E, Sassi F, Noghero A, Erriquez J, Perera T, Olivero M, Di Renzo MF.Heat-shock protein 27 (HSP27, HSPB1) is up-regulated by MET kinase inhibitors and confers resistance to MET-targeted therapy. FASEB J. 2014, 28:4055-67.


Maria Flavia Di Renzo got an MD degree at the University of Torino Medical School in 1977 and then the resi-

dency in Neurology in 1981. Besides short periods spent working in another University and abroad, she has

been working in the same University as professor since 1981 and as full professor of Histology since 1994 to

date. In 1997 she became head of the Laboratory of Cancer Genetics of the Candiolo Cancer Institute.

Since 1981 she has been involved in the study of oncogenes and tumor suppressor genes in human cancer with

a particular focus on tyrosine kinase oncogenes, demonstrating for the first time the increased expression of

the MET oncogenes in human cancer (Oncogene, 1991). Moreover, she identified mutations of the MET oncoge-

ne in lymph node metastases of head and neck squamous cell carcinomas, reinforcing the concept of a genetic

basis of the cancer progression towards metastasis.

In 2004 her major research interest turned in the study of human ovarian cancer with the aim of identifying su-

itable targets for molecular therapy. For this purpose the group led by Maria Flavia Di Renzo has used functional

(Cancer Res 2004; Clin. Cancer Res 2007), genomic (Mol. Cancer Ther., 2006; FASEB J 2012) and proteomic

approaches (FASEB J and J Proteome Res 2014), that have led to the identification of possibly actionable

molecules.


Cancer stem cells and resistance to standard and targeted therapiesTo set up therapies targeted at ‘cancer stem cells’ that constituite the ‘roots of tumors’. These specific cells may resist chemo- and radio-therapy, and, after these conventional treatments, can cause tumor relapse

TeamFrancesca De Bacco, Paolo Luraghi, Federica Verginelli, Viola Bigatto, Antonio D’Ambrosio, Francesca Orzan, Raffaella Albano, Elena Casanova, Elia Cipriano, Roberta Neggia, Gigliola Reato

Research topic:

Cancer stem cells and resistance to standard and targeted therapies

Background:

Like normal tissues, tumors such as colorectal cancer or glioblastoma are structured according to a hierarchy that

includes two main components. At the hierarchy’s apex there is a (small) subpopulation of ‘cancer stem cells’ (CSC)

able to self-renew, namely to endlessly propagate, and support tumor histogenesis, regeneration, and dissemination.

At the hierarchy’s base there is an (ample) subpopulation of cells that, unlike CSC, have limited self-propagation abi-

lity, and tend to aberrantly differentiate and die. This model implies that, to cure the tumor, CSC must be eradicated.

This goal is challenging, as CSC are often inherently resistant to therapies that so far have proven effective against

the hierarchical basis of tumor cells, therapies that also include the most innovative agents, targeted at the genetic

alterations driving cancer pathogenesis. To improve therapeutic outcomes, it is mandatory to identify and attack the

molecular and genetic mechanisms underlying CSC resistance to conventional chemo- and radio-therapy, or to agents

targeted at molecular lesions.


In CSC isolated from glioblastoma (GBM-CSC), we have shown that the wild-type MET oncogene (the HGF receptor)

is a functional marker of a specific subset of CSC, it activates a signalling pathway that sustains the DNA damage

response, and counteracts cell death induced by ionizing radiation. As result, MET inhibition sensitizes GBM-CSC to

radiotherapy in vitro and in vivo, and promotes regression of tumors obtained by GBM-CSC transplantation throu-

gh eradication of the radioresistant GBM-CSC subpopulation. In CSC isolated from metastatic colorectal cancer

(CRC-CSC), we’ve shown that several growth factors, including HGF and members of the EGF family, counteract the

proliferative blockade imposed by EGFR inhibitors, currently used to treat metastatic CRC with significant but incom-

plete success. In mouse models obtained by CRC-CSC transplantation, we’ve shown that the therapeutic efficacy of

standard EGFR inhibition is greatly enhanced by concomitant inhibition of MET and multiple members of the EGFR

family.


The MET oncogene and EGFR, are both essential to sustain CSC viability and long-term propagation in tumors such

as GBM and CRC. In GBM, where MET and EGFR are often expressed in a mutually exclusive manner, we’ve provided

preclinical evidence that MET targeting impairs CSC properties, and increases the effectiveness of current therapies

aimed at affecting DNA integrity and cell survival (e.g. radiotherapy). In CRC, we dissected the relative contribution of

the four members of EGFR family and MET to CSC propagation and tumorigenicity. These results can instruct clinical

trials that associate MET or EGFR family inhibitors with standard therapeutic protocols. Current work is aimed at

identifing (i) the genetic and molecular context that modulates the CSC response to MET and EGFR inhibitors; (ii) the

intratumoral CSC genetic and phenotypic heterogeneity, which must be recognized to refine the targeted treatment

of each individual tumor.

Selected references


Can

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ancer In

stitute

Carla Boccaccio MDLaboratory of Cancer Stem Cells Research



Fax. +39.011.9933225

Boccaccio C, Luraghi P, Comoglio PM. MET-mediated resistance to EGFR inhibitors: an old liaison rooted in colorectal cancer stem cells. Cancer Res. 2014. 74:3647-51

Luraghi P, et al. MET signaling in colon cancer stem-like cells blunts the therapeutic response to EGFR inhibitors. Cancer Res. 2014, 74:1857-69

Boccaccio C and Comoglio PM. The MET oncogene in glioblastoma stem cells: implications as a diagnostic marker and a therapeutic target. Cancer Res. 2013, 73:3193-9.


From the beginning of her career, Carla Boccaccio has contributed to elucidate the signalling and transcriptio-

nal mechanisms by which the MET oncogene controls invasive growth, a physiological program of stem and

progenitor cells, inappropriately activated during tumor onset and progression (nature Rev Cancer, 2006).

She discovered that MET regulates a genetic program concomitantly driving a procoagulant and a neoplastic

phenotype (Nature, 2005), thus helping to unravel the pathogenesis of Trousseau’s syndrome, the association

of cancer and thrombosis frequently observed, but often unexplained, in patients (J.Clin.Oncol, 2009). More

recently she discovered that MET is a functional marker of cancer stem cells isolated from brain and colorectal

tumors (Cancer Res, 2012). In these cells, MET sustains self-renewal, invasion, and resistance against DNA da-

maging treatments, such as ionizing radiation, and biological agents targeting other tyrosine kinase receptors

such as EGFR. She is now engaged in understanding the interactions between MET activity, the genetic lesions

occurring in cancer stem cells, and the cellular response to standard and targeted treatments, in order to iden-

tify the clinical context where MET inhibition can be effective (EMBO Mol.Med.,2015)


Integrative genomics of cancer progression and resistance to treatmentsThe integration of molecular profiles and clinical information enables accurate determination of cancer aggressive-ness and response to treatment and to individualize therapeutic approaches (precision medicine)

TeamClaudio Isella, Consalvo Petti, Gabriele Picco, Sara Erika Bellomo, Andrea Mignone, Federica Invrea

Research Topic:

Integrative genomics of cancer progression and resistance to treatment.

Background:

Cancer progression involves acquisition by neoplastic cells of several molecular alterations, conferring meta-

static ability and resistance to anticancer treatments. Integrative genomics evaluates systematically, in cohorts

of tumor samples, multiple layers of molecular information, at the DNA, mRNA and microRNA level. The same

analyses are conducted on large series of preclinical models, such as cell lines and patient-derived xenografts

(PDXs), to consolidate regulatory network alterations in individual cancers and cancer subtypes, to identify key

lesions driving cancer progression and possibly to determine sensitivity to targeted treatments. Extensive vali-

dation in the preclinical models allows exploiting the genes as therapeutic targets, and molecular signatures as

diagnostic tools for precision medicine.


The Laboratory of Oncogenomics has been focusing on colorectal cancer (CRC) and on the generation and

analysis of multi-layer molecular and pharmacological data on large sets of CRC samples, patient-derived xe-

nografts (PDXs) and cell lines. Such analysis was then followed by functional validation of identified candidates

and mechanisms of neoplastic progression. The main achievements are here summarized: (i) species-specific

genomics of CRC PDXs to distinguish the stromal transcriptome, contributed by murine cells, from the human

epithelial cancer transcriptome, with implications in CRC aggressiveness and resistance to treatment (Nature

Genetics, 2015); (ii) identification of mixed mRNA/microRNA networks associated to CRC subtypes (Nature

Commun., 2015) and of multi-dimensional networks to uncover cancer drivers (Scientific Reports, 2015); (iii)

Integrative pharmacogenomic profiling of a large collection of CRC cell lines (n=151), validating molecular

subtypes and providing new actionable expression outliers in specific cases (Nature Commun., 2015) (iv) De-

velopment of a procedure for high efficiency in vivo transduction of PDXs with lentiviral vectors (Scientific Re-

ports, 2015); (viii) Identification of NEDD8 pathway inhibition as an effective therapeutic option in KRAS- and

BRAF-mutant CRC, and in vivo validation of a molecular response predictor (JNCI, manuscript submitted). Sha-

ring the genomic expertise of the Lab with other groups inside and outside the Institute also led to publication

of several collaborative works.

Conclusions and Perspectives:

The Laboratory will continue exploiting integrative genomics for better molecular stratification and for highligh-

ting actionable therapeutic targets in CRC and other neoplastic diseases.

Selected references


Can

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Enzo Medico MDLaboratory of Oncogenomics



Fax. +39.011.9933225

Isella C., et al. Stromal contribution to the colorectal cancer transcriptome. Nat Genet. 2015, 47(4):312-9.

Cantini L, Isella C, Petti C, Picco G, Chiola S, Ficarra E, Caselle M, Medico EMicroRNA–mRNA interactions underlying colorectal cancer molecular subtypes. Nat Commun. 2015, 6: 8878.

Medico E, et al. The molecular landscape of colorectal cancer cell lines unveils clinically actionable kinase targets. Nat Commun. 2015, 6:7002


Enzo Medico graduated in Medicine at the University of Torino in 1989 and got a PhD in Cell and Developmental

Biology in 1994, University “La Sapienza” of Rome. Since 2005 he is associate professor at the University of

Torino. Dr. Medico has a long-standing activity in the field of tyrosine kinase receptor signaling and genomics.

In particular, he has worked on the MET family of tyrosine kinase receptors, showing their role in the control of

epithelial morphogenesis and invasive growth. Since 1998, Dr. Medico has re-directed his research in the field

of functional genomics. In collaboration with the Fred Hutchinson Cancer Research Center, Seattle, he develo-

ped a “gene trapping” approach for genome-wide exploration of transcriptional regulation. Dr. Medico has been

among the first Italian users of DNA microarrays technology, exploiting it since 1999 to study alterations in

transcriptional regulation, gene expression and function associated with cancer aggressiveness and resistance

to treatment. In 2004 he became director of the Laboratory of Oncogenomics at the Candiolo Cancer Institute.

Over the last ten years, he has been managing and coordinating functional oncogenomics projects led by the In-

stitute and funded, among others, by the Italian Ministry of Health, by AIRC and by the European Commission.


The knowledge of specific molecular alterations in the tumor DNA repair pathway could define a novel context of susceptibility for alkylating agents

Pharmacogenomics of colorectal cancer

Team Ludovic Barault, Daniele Oddo

Research topic:

Biomarkers for personalized colorectal cancer therapy

Background:

Colorectal cancer patients often show variable responses to treatment. This variation is the result of the mo-

lecular heterogeneity underlying the complex biology of cancer cells. Our main interest is therefore focused in

understanding the changes that occur in tumor cells on exposure to anti-cancer agents. Our current programme

concentrates on projects aimed at determining biomarkers of response or resistance to anti-cancer treatments

with the ultimate goal of designing and testing sequences of drugs rationally designed on the modifications

induced by the first agent on the tumor molecular landscape.


As a model, we focused on BRAF mutant colorectal cells with acquired resistance to target therapies combina-

tions. Genotyping of resistant cells identified heterogeneous molecular mechanisms of acquired drug resistan-

ce that converge on reactivating MAPK signaling. Drug resistance could be overcome by ERK inhibitor-based

or other targeted therapy combinations, based upon the selective target(s) identified in resistant cells. Of

translational relevance, we demonstrated how the combined data obtained by tumor molecular profiling and

functional experiments in cell lines informed physician’s choice of administering a specific sequential regimen,

which induced clinical benefit in an individual BRAF mutant colorectal cancer patient.


This approach led us to define sequential strategies suitable for clinical testing in BRAF mutant colorectal can-

cer patients. As a future perspective, we intend to study how tumor preconditioning with epigenetic drugs could

influence the response of BRAF mutant malignancies to subsequent anti-cancer therapies.

Selected references


Can

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Federica Di Nicolantonio PhDLaboratory of Cancer Epigenetics

University of Torino Medical SchoolE-mail: [email protected]

Phone. +39.011.9933827 Fax. +39.011.9933225

Barrault L., et al. Digital PCR quantification of MGMT methylation refines prediction of clinical benefit from alkylating agents in glioblastoma and metastatic colorectal cancer. Annals of oncology. 2014 ; 26: 1994-9

Sun C, et al. Reversible and adaptive resistance to BRAF(V600E) inhibition in melanoma. Nature. 2014 3;508:118-22. Zecchin D, et al. BRAF V600E Is a Determinant of Sensitivity to Proteasome Inhibitors. Mol Cancer Ther. 2013; 12:2950-61.


Federica Di Nicolantonio holds a PharmD degree (1999) and gained a PhD (Medicine) from University College

London, UK, in 2004. Her research focuses on the molecular mechanisms underlying cancer cells’ response

to antineoplastic drugs. She identified KRAS mutations and KRAS amplification as biomarkers of resistance

to the mTOR inhibitor everolimus (J Clin Invest. 2010) and EGFR monoclonal antibodies (Int J Cancer 2013),

respectively. She first described BRAF mutations as a biomarker of adverse prognosis and of resistance to

EGFR targeted therapies in colorectal cancer patients (J Clin Oncol. 2008) and discovered that lack of efficacy

of BRAF inhibitors could be mediated by EGFR (Nature 2012). This preclinical work has provided the rationale

for testing BRAF and EGFR inhibitors in colorectal cancer patients.

EGFR MET KRAS BRAF

After treatment with combinated BRAF+EGFR inhibitors

Baseline

At progression

FISH analysis on chromosome metaphase spreads shows oncogene amplifications in distinct colon cancer cell subpopulations with acquired resistance to BRAF and EGFR inhibitors.


Personalized therapy and non-invasive molecular diagnostics of colorectal cancerTo define the genetic and molecular characteristics of patients in order to lead and check the treatment of colorectal cancer

Team Sabrina Arena, Giovanni Crisafulli, Giovanni Germano, Sebastian Hobor, Simona Lamba, Luca Lazzari, Sandra Misale, Mariangela Russo, Beth Van Emburgh, Giulia Siravegna, Carlotta Cancelliere, Benedetta Mussolin, Luca Novara, Giuseppe Rospo

Research topic:

Clonal evolution and drug resistance: from cancer avatars to liquid biopsies.

Background:

A central paradigm of modern oncology is that tumor-specific molecular alterations underlie ‘functional’ depen-

dencies that can be therapeutically exploited. Remarkable results have been obtained by applying this paradigm

in the clinic, and several targeted drugs have been approved to treat, among others, melanomas, lung and co-

lorectal cancers. Unfortunately, when metastatic cancers are challenged with targeted agents almost invariably

a subset of cells insensitive to the drug emerges. As a result, in most instances, targeted therapies are only

transiently effective in patients. Arguably, the rapid and apparently unavoidable emergence of resistance is the

major limitation to further progress in the application of targeted therapies in oncology. How can we overcome

the near-certainty of disease recurrence following treatment with targeted agents?


We have used colorectal cancer (CRC) as a model system to test the hypothesis that the emergence of drug resi-

stance can be controlled. To this goal, we studied how clonal evolution impacts the development of resistance in

patients’ avatars (cell lines and tumor xenografts). In parallel, we analysed clonal evolution dynamics and drug

resistance in the blood of patients (liquid biopsies) who received targeted therapies. Our results indicate that

acquired resistance is -for the most part- a ‘fait accompli’. Mathematical models based on our data suggest that

each (traceable) metastatic lesion contains hundreds of cells with mutations to virtually any targeted agents.

According to this model, the time to relapse is simply the interval required for mutant or otherwise altered sub-

clones to re-populate the lesion. On the positive side, we found that resistance can be prevented when drug

combinations are rationally used.


The awareness that in nearly all patients targeted drugs are only transiently effective due to the rapid emer-

gence of resistance poses a formidable challenge. Although the overall picture is looming, our data and the

concepts they underlie offer several opportunities for intervention. Genetic tracking of cancer cell populations

can be used to study how the development of resistance can be restrained in vitro and in vivo. Furthermore,

liquid biopsies can be exploited to monitor clonal evolution and to intercept the emergence of resistant clones

non-invasively in patients. When applied together cancer avatars and liquid biopsies may identify, monitor, and

potentially overcome resistance to targeted therapies.

Selected references


Can

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Alberto Bardelli PhDLaboratory of Molecular Oncology



Fax. +39.011.9933225

Siravegna G.,et. al. Clonal evolution and resistance to EGFR blockade in the blood of colorectal cancer patientsNat Med. 2015 Jul;21(7):795-801

Arena S., et al. MM-151 overcomes acquired resistance to cetuximab and panitumumab in colorectal cancers harboring EGFR extracellular domain mutations. Sci Transl Med. 2016 Feb 3;8(324):324ra14.

Russo M. et al. Acquired resistance to the TRK inhibitor entrectinib in colorectal cancer Cancer Discov. 2016 Jan;6(1):36-44


A. Bardelli received a PhD in Molecular Biology from the UCL (University College London, UK). As a post doc

at the Johns Hopkins University (Baltimore, USA, supervisor Dr. Bert Vogelstein) he developed the first com-

prehensive mutational profile of kinases (Science 2003). As an independent investigator, A. Bardelli has then

translated these findings into clinical practice (Lancet Oncology 2005; JCO 2008). Recently, he has defined

mechanisms of acquired resistance to anti EGFR therapies in colorectal cancer patients thus providing insights

into new therapies aimed at overcoming resistance (Nature 2012; Cancer Discovery 2014). These studies invol-

ve an innovative methodology - named liquid biopsy - which allows the use of circulating tumor DNA to monitor

patient’s response using a blood draw (Science Translational Medicine 2014).


Targeted cancer immunotherapyDevelopment of novel therapeutic strategies to generate, produce, and deliver monoclonal anti-cancer antibodies

TeamSimona Cignetto, Cristina Chiriaco, Lara Fontani

Research topic:

Development and validation of cancer immunotherapy strategies based on MET antibody and MET antibody-derived

molecules.

Background:

Clinical evidence indicates that the MET oncogene plays a role in progression of cancer toward metastasis and/or

resistance to targeted therapies. While mutations are rare, the common mechanism of MET activation is overexpres-

sion, either by gene amplification (‘addiction’) or transcriptional activation (‘expedience’). In both instances ligand-

independent kinase activation plays the major role in sustaining the transformed phenotype. Currently available MET

antibodies are directed against the receptor binding site, behaving essentially as a ligand (HGF) antagonist, and are

ineffective in ligand-independent activation.


The monovalent chimeric MvDN30 antibody, delivered as a purified protein, binds the extracellular domain and indu-

ces the proteolytic cleavage of MET, dramatically inhibiting downstream signaling pathways, in both the absence or

presence of ligand. MvDN30 has the limit of poor in vivo stability. To address this issue MvDN30 was delivered by

‘gene therapy’, to induce a stable and constant in vivo production that counterbalances the high rate of clearance. As

alternatives, the molecule has also been chemically modified (i.e. Pegylated) or engineered, including two repetitions

of the constant domain (DCD). All the explored strategies led to the improvement of the MvDN30 potency. By the gene

therapy approach, direct Mv-DN30 gene transfer in nude mice, intra-tumor or systemic, was followed by a therapeutic

response of MET addicted human glioblastomas and lung carcinomas. Upon Pegylation the antibody overcame the

resistance to EGFR targeted therapy in a MET-amplified patient-derived colorectal tumor (xenopatient). The DCD appro-

ach was successfully applied to inhibit the peritoneal carcinomatosis of a MET-addicted gastric tumor.


We developed strategies suitable to improve the potency of MvDN30, rendering it an attractive candidate for clinical ap-

plications. To further enlarge the panel of anti-MET immunotherapy routes we aim to explore a strategy of re-directed

adoptive immune response by a Chimeric Antigen Receptor anti-Met.

Selected references


Can

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ancer In

stitute

Elisa Vigna PhDLaboratory of Gene Transfer

Research AssociateUniversity of Torino Medical School


Fax. +39.011.9933225

Vigna E, Pacchiana G, Chiriaco C, Cignetto S, Fontani L, Michieli P, Comoglio PM. Targeted therapy by gene transfer of a monovalent antibody fragment against the met oncogenic receptor. J Mol Med (Berl).2014, 92:65-76.

Vigna E, Comoglio PM. Targeting the oncogenic met receptor by antibodies and gene therapy. Oncogene. 2015, 34:1883-1889.

Vigna E, Chiriaco C, Cignetto S, Fontani L, Basilico C, Petronzelli F and Comoglio PM. Inhibition of ligand-independent constitutive activation of the met oncogenic receptor by the engineered chemically-modified antibody dn30. Molecular Oncology. 2015, 9: 1760-1772.


Elisa Vigna was involved in the identification of HGF/SF as the Met ligand (Oncogene, 1991; EMBO J., 1991) and in the

characterization of the biological activation of HGF by urokinase (EMBO J., 1992; J Biol Chem, 1995). She studied the

role of the Met juxtamembrane domain in the regulation of Met activity (Oncogene). In the gene therapy field

she acquired experience collaborating with Prof. Naldini, who conceived and developed the Lentiviral Vectors.

She performed pionieristic ‘vectorology’ studies optimizing lentiviral vectors for multiple gene transfer and

exogenously regulated expression (Mol. Therapy, 2002; Mol. Therapy, 2005; Nature Biotech, 2005). She cha-

racterized the anti-Met antibody DN30 and generated its monovalent non-agonistic counterpart. She provided

proof of concept for either conventional or ‘gene transfer based’ protocols for DN30 application (Cancer Res,

2008; J Biol Chem, 2010; J Mol Med 2014).

(A) Anti-Met gene therapy on Glioblastoma cells treated with lentiviral vector expressing the monovalent Met antibody DN30. The treatment significantly improves overall survival. (B) Anti-Met targeted therapy on a patient derived ‘Met-addicted’ colo-rectal cancer (‘xenopatient’). The monovalent chemically stabilized Met antibody DN30 in combination with the Epidermal Growth Factor Receptor antibody (Cetuximab) 62% reduces tumor growth.(C) Anti-Met targeted therapy on Met-addicted gastric carcinoma cells. The engineered recombinant DCD molecules derived from the Met antibody DN30 45% reduced peritoneal carcinomatosis.


Mechanisms of resistance to tyrosine kinase receptor inhibitors

TeamSimona Corso, Elena Ghiso, Valentina Martin, Silvia Menegon, Cristina Migliore, Annalisa Petrelli, Maria Apicella, Marilisa Cargnelutti, Elena Morando

Design of strategies bypassing resistance to targeted therapies

Research topic

Altered regulation of tyrosine kinase receptors is frequent in solid tumors and it is often associated with the acquisition

of an aggressive phenotype. Thus, therapies targeting these receptors have been proposed as molecular approaches

to treat human cancers. The main problem with these therapies is the onset of resistance, which leads to treatment

failure. It is therefore important to understand the mechanisms of resistance to molecular targeted therapies, in an

effort to optimize the outcome of the treatments.

Background:

The goal of delivering the right drug to the right cancer patient (precision medicine) requires a detailed understanding

of how genomic alterations are linked to drug response. Hence, the clinical efficacy of target therapy depends on: i) the

validation of the true biological relevance of putative targets in the context of a specific tumor type; ii) the context-specific

identification of the active drug(s) and of additional molecular alterations affecting the responsiveness to treatment.


Our group aims to study the molecular mechanisms that allow tumor cells to become insensitive to molecular targeted

therapies. We focused on the study of gastric cancer that is the world’s third leading cause of cancer mortality We

generated a platform of more than 70 patient-derived xenografts (PDXs) that recapitulate the heterogeneity of this

disease. We aim to genetically dissect the gastric tumor landscape in order to investigate mechanisms involved in

sensitivity/resistance to targeted therapies against receptor tyrosine kinases.

We performed the genomic analysis of the PDX tumors and we identified several alterations of tyrosine kinases such as

HER2, EGFR, MET, FGFR2 and of KRAS. We exploited a MET-amplified PDX model to optimize anti-MET therapeutic stra-

tegies in gastric cancer. We report that despite the high MET amplification level (26 copies), tumors showed only partial

and transient sensitivity to anti-MET therapy while dual MET/EGFR inhibition led to complete regression and prevented

resistance onset. The finding that combined anti-MET/EGFR therapy even in the absence of EGFR genetic alterations

induced complete and durable response, represents a proof of concept and guarantees further investigations, opening a

new perspective of treatment for these patients. We also performed pre-clinical trials on several HER2-amplified gastric xe-

nopatiens to identify the best anti-HER2 therapeutic approach and investigate the mechanisms of resistance to treatment.


By exploiting the gastric cancer PDX platform we have generated, we aim to unravel genetic vulnerabilities in this pa-

thology and to identify the best therapeutic strategies to target them. Since gastric cancer is a very common disease

worldwide, the identification of novel ‘druggable’ and validated targets would be extremely important from a clinical point

of view, even if their prevalence is very low. As a whole, the results of this project will provide a scientific basis for future

clinical applications and guide the rational design of molecularly-oriented clinical trials for gastric cancer.

Selected references


Can

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Silvia Giordano MD, PhDLaboratory of Molecular Biology

Full ProfessorUniversity of Torino Medical School


Fax. +39.011.9933225

Bardelli A., et al. Amplification of the met receptor drives resistance to anti-egfr therapies in colorectal cancer. Cancer discovery, 2013, 3:658-73

Corso S., Giordano S. Cell-autonomous and non-cell-autonomous mechanisms of hgf/met-driven resistance to targeted therapies: from basic research to a clinical perspective. Cancer discov. 2013, 3:978-92

Zavattari P, et al. Nrf2, but not ß-catenin, mutation represents an early event in rat hepatocarcinogenesis. Hepatology. 2015 62:851-62


Silvia Giordano has a long experience in the field of research on tyrosine kinase receptors and related onco-

genes. She has focused her research on the study of the tyrosine kinase receptor encoded by the MET gene.

She identified this receptor (Nature, 1989) and contributed to the understanding of the signal transduction

mechanisms and of the biological role in physiological as well as in pathological conditions. In particular, she

has critically contributed to the understanding of the mechanisms controlling MET down-regulation (Nature,

2002) and HGF-independent activation (Nature Cell Biol., 2002). Moreover, she showed that cancer cells may

be dependent (“addicted”) on the constitutive activation of this oncogene (Oncogene, 2008). Recently, she has

been deeply involved in the study of the mechanisms of resistance to targeted therapies (Cancer Res., 2010;

Cancer Discovery, 2013). She has also performed studies of the role of microRNAs in cancer progression (Can-

cer Res., 2008; Clin. Cancer Res., 2012; Hepatology, 2014). On the translational front she has identified a MET

monoclonal antibody (DN30) able to impair Met activation and tumorigenicity (Proc Natl Acad Sci U S A.,2006).


Research topic:

Immunotherapy with Cytokine-Induced Killer (CIK) cells for the treatment of solid tumors refractory to conven-

tional treatments.

Background:

Adoptive immunotherapy holds promise for the treatment of solid tumors. Disease relapse and drug-resistance

are sustained by a subset of putative cancer stem cells (CSC), poorly responsive to conventional treatments.

Immunotherapy is a promising approach capable of targeting putative CSC. Our group has focused over the last

years on a subset of T lymphocytes known as Cytokine-induced Killer (CIK) cells. CIK cells are ex vivo expanded

T lymphocytes generated from circulating mononuclear cells, endowed with MHC-independent antitumor activi-

ty. The aim of our research is to explore adoptive immunotherapy with CIK cells for sarcomas and melanomas.


We demonstrated that immunotherapy with CIK cells is active against autologous sarcomas and melanomas,

including putative CSC. Our main findings are summarized as follows. (i) Generation of CIK cells: clinically re-

levant rates of CIK cells were successfully expanded from patients with sarcomas and melanomas; (ii) tumor

killing activity: patient-derived CIK cells were highly active against autologous sarcomas and melanomas pre-

treated with conventional chemotherapy; (iii) activity against CSC: we developed a gene-transfer strategy capa-

ble of visualizing putative CSC based on their ability to re-activate stem gene Oct4. Sarcoma and melanoma

CSC were relatively resistant to conventional chemotherapy but susceptible to immunotherapy with CIK cells;

iv) In preliminary experiments we demonstrated the possibility to redirect CIK cells with a chimeric-antigen

receptor (CAR) against a precise target expressed in soft tissue sarcomas.


Adoptive immunotherapy with CIK cells is potentially active against solid tumors and capable of eradicating

chemoresistant CSC. We plan to enhance the antitumor activity of CIK cells by their genetic redirection against

tumor-specific targets. Furthermore we will explore synergism of immunotherapy with molecular targeted ap-

proaches.

TeamLoretta Gammaitoni, Lidia Giraudo Diego, Giulia Mesiano, Valeria Leuci

Experimental cancer immunotherapy with Cytokine-induced killer cells

Adoptive immunotherapy against solid tumors

Selected references


Can

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Dario Sangiolo is a physician scientist with research interest in the field of transplant immunology developed

at the Fred Hutchinson Cancer Research Center (Seattle, WA). He contributed to the development and characte-

rization of preclinical models based on Cytokine-Induced Killer (CIK) cells (Int Immunol. 2008; J Immunother.

2012). In preclinical studies he demonstrated the activity of CIK cells against melanomas and sarcomas (Clin

Cancer Res. 2013; Cancer Res. 2014 ). He explored strategies to genetically engineer T lymphocytes to enhance

antitumor activities and decrease the risk of GVHD across HLA barriers (Hum Gene Ther. 2009; Gene Ther.

2007). In the field of clinical research he investigated the efficacy and toxicity of Hematopoietic Cell Transplant

in hematologic and solid tumors (Biol Blood Marrow Transplant. 2010; Blood. 2006 May).

Grignani et al. Sorafenib and everolimus for patients with unresectable high-grade osteosarcoma progressing after standard treatment: a non-randomised phase 2 clinical trial. Lancet Oncology. 2015, 16:98-107

Sangiolo D. et al. Cytokine-induced killer cells eradicate bone and soft-tissue sarcomas. Cancer Research. 2014, 74:119-29

Gammaitoni et al. Effective activity of cytokine-induced killer cells against autologous metastatic melanoma including cells with stemness features. Clinical Cancer Research. 2013, 19:4347-58

Dario Sangiolo MDUnit of Experimental Cell Therapy



Fax. +39.011.9933290

52 | Investigational Clinical Oncology

The Heracles Clinical Trial

Investigators: Alberto Bardelli, Silvia Marsoni, Salvatore Siena, Silvia Giordano, Carla Boccaccio, Paolo Mi-

chieli, Livio Trusolino, Paolo M.Comoglio - Scientific Advisors: Marc Ladanyi (Memorial Sloan Kettering Cancer

Center, New York, US), Manuel Hidalgo (Spanish National Cancer Research Centre, Madrid, ES), David Ran-

sohoff (UNC Lineberger Comprehensive Cancer Center, Chapel Hill, US)

Investigational Clinical Oncology | 53

Research Topic 3Investigational Clinical Oncology

Over the past few years, the improved knowledge on the biological, genetic and molecular heterogeneity of tumors, together with the development of pharmacological technologies, has allowed the identification of molecular targets for novel therapeutic strategies. This fast process has led to the overall reconsideration of classical approaches to clinical oncology, traditionally oriented toward design of cancer treatments irrespective of the biological and genetic peculiarities that can make each tumor a pathology on its own. The identification of patients likely to respond to specific treatments according to the presence of relevant molecular targets (personalized medicine) and based on the expression of potential markers of sensitivity or resistance, needs clinical studies that result from a constant and productive interaction among the professionals with a significant background in the various disciplines. The goal of the “Investigational Clinical Oncology” team is to increase the therapeutic index of molecular targeted drugs, by the identification and clinical validation of biomarkers of sensitivity/resistance. This goal is pursued through the development of research programs aimed at designing hypothesis-driven clinical trials that directly derive from the Institute’s biological, genetic and molecular research. In order for these tasks to be implemented, an ad-hoc organizational structure favours the interchange and the synergy among preclinical researchers, oncologists and pharmaceutical companies – which are the potential providers of new drugs for clinical trials. Another goal of INCO will be the promotion of a clinical research culture based on the knowledge of molecular biology and tumor genetics, and particularly of the mechanisms of sensitivity or resistance to molecular targeted drugs. This organization is instrumental to the referral of patients for admission into top-priority clinical trials.

Research Topic 3 is focused on:3.0 Rational design of clinical trials for targeted drugs


HERACLES and ARES: trials for the ‘Precision Medicine’ of Cancer

Team Antonella Balsamo, Cosimo Martino, Marilì Vitiello, Mario Spione

Shaping the clinical validation path of new putative targets found in the Institute’s preclinical programs

Research topic:

Design, implementation and conduction of ‘hypothesis-driven’, precision medicine trials based on the transla-

tional discoveries of the FPO-IRCC research laboratories

Background:

Precision medicine brings a new set of challenges to clinical application. To hasten the adoption of new thera-

pies we need to achieve a deeper molecular understanding of cancers and reliably translate results from repre-

sentative preclinical models into clinical trials enriched with suitably identified patients.


At Candiolo we have started to confront precision medicine hurdles by creating a clinical platform integrating,

under the same virtual roof, the efforts of surgeons, pathologists, radiologists, and medical oncologists from

the IRCC clinical network (25 centers around Italy), in order to assemble the timely collection of high quality,

clinically annotated biological samples at meaningful time points, to feed research projects and precision me-

dicine trials. Five translational protocols are now feeding the translational Labs of the Institute (PROFILING,

AGNOSTOS-Profiling, GEA, FUNNEL and CORNUCOPIA). Together with the translational scientists, we are also

building a “cancer knowledge network” to store the resulting molecular and medical data in digital form and

to deliver them, in comprehensible ways, to scientists, clinicians, and patients (Projects LAS and PROBUS).

Three hypothesis driven clinical trials have been designed and implemented (HERACLES and ARES in colorec-

tal cancer, AGNOSTOS in cancer of unknown origin). HERACLES has been the the Institute first precision trial

designed upon the discovery, based on PDX studies, that HER2 amplification in metastatic colorectal cancer

(mCRC) associates with resistance to anti-EGFR therapy, and predicts response to the combination of lapatinib

and trastuzumab for a dual blockade of the HER2 pathway. Accordingly, we conducted a proof-of-concept trial

testing the combination in HER2 amplified mCRC patients failing standard therapies. The trial has proven the

efficacy of the combination with 8 long lasting responders out of 23 patients treated, proving HER2 as the first

bona fide actionable target in mCRC. Genotype/response correlation has shown that only patients with truly

‘HER2 addicted’ tumors benefit from this therapy. The tumor genomic landscape of non-responders and relap-

sing cases is under scrutiny to define the determinants of primary and secondary resistance.


The obstacles encountered in HERACLES will be addressed: unexplained primary drug resistance, inadequacy

of current criteria for monitoring tumor response and recurrence, limited knowledge of genomic heterogeneity

of mCRC. We will also extend the dynamic research infrastructure which we have created for mCRC to gastric

cancer and cancers of unknown origin. Two new resource-dense, hypothesis-driven trials deriving their rationale

from the results of the Institute’s translational labs, will be embedded and nurtured by this infrastructure.

Selected references


Can

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Silvia Marsoni MDClinical Trial Coordination Unit (CRO)


Fax. +39.011.9621525

Bergmann L, et al. Actual developments in European regulatory and health technology assessment of new cancer drugs: what does this mean for oncology in Europe? Ann Oncol. 2014, 25:303-6.

Garassino MC, et al. TAILOR trialists. Erlotinib versus docetaxel as second-line treatment of patients with advanced non-small-cell lung cancer and wild-type EGFR tumours (TAILOR): a randomised controlled trial. Lancet Oncol. 2013, 14:981-8.

De Marinis F, et al. A phase II study of the histone deacetylase inhibitor panobinostat (LBH589) in pretreated patients with small-cell lung cancer. J Thorac Oncol. 2013, 8:1091-4


Silvia Marsoni has a distinguished career as a clinical trial methodologist, which started in the ’80 at the NCI-

USA. Since then, she has focused her research efforts on the rationalization of oncology trials. In the nineties at

the Mario Negri Institute she founded her first clinical network and designed pivotal phase 3 trials establishing

the standard of care in the adjuvant setting of colon cancer. Currently, she still designs standard-defining trials

for genotype selected populations. In 2000 she founded SENDO, a non-profit organization for the early deve-

lopment of new cancer drugs in Italy and Spain. SENDO, firstly in Italy, developed targeted drugs in biomarker

enriched populations. While there she also developed trabectidin in ovarian cancer. Since 2011 she directs the

Clinical Trials Unit, and has built the clinical platform for genomic-driven trials serving the translational labs of

the Institute. The first of these trials, HERACLES, is targeting HER2 in colorectal cancer with promising preli-

minary results.


Cancer of Unknown Primary (CUP): the AGNOSTOS PROJECT

TeamGiulia Maria Stella, Elena Geuna, Andrea Milani, Annamaria Nuzzo, Giorgia Zucchini, Marilisa Cargnelutti, Alice Maria Balderacchi

Research topic:

Cancer of Unknown Primary (CUP): the AGNOSTOS PROJECT

Background:

Cancer of Unknown Primary (CUP) is a heterogeneous clinical and pathological syndrome characterized by

early tumor metastatic spread without an identifiable, site-specific primary tumor of origin. Despite accounting

for 3-5% of all human cancers, CUP represents a still unsolved clinical problem, with a not fully understood

biology and paucity of effective therapies. Currently, patients with CUP are given systemic chemotherapy, based

on a semi-agnostic approach that considers general histological features rather than organ-derivation. With 25-

30% response rate and overall survival ranging from 6 to 16 months, current treatment of CUPs is considered

largely suboptimal. Furthermore, because of their innate ability to spread at distant organs early in their natural

course, CUPs may represent a valuable clinical and translational model to study the molecular basis of tumor

metastatization.

Research Achievements:

We have designed and implemented the AGNOSTOS program, that is aimed at identifying new potential tre-

atments and at conducting systematic translational studies on this rare entity. The AGNOSTOS program inclu-

des two prospective clinical studies.

• The AGNOSTOS 1 study is a multicenter, randomized phase 2 trial with a ‘Pick the Winner’ design to assess the

efficacy of nab-paclitaxel-based doublet as first line therapy in treatment-naïve CUP patients. In particular we

will explore the activity of nab-paclitaxel in combination with the two most active drugs (carboplatin or gemci-

tabine). The trial is ethically approved and currently ongoing in 10 Italian Institutions under the coordination of

the Candiolo Cancer Institute. We plan to enroll 60 patients in each arm of the trial.

• The AGNOSTOS Companion Translational study is aimed at the collection of vital tissues from CUP patients

to establish patient-derived-xenografts (PDX) and cancer stem cells (in vitro). Furthermore we aim to collect

blood/plasma from CUP patients to obtain free tumor DNA from plasma to follow tumor burden and clonal

evolution in CUPs patients (liquid biopsy), and circulating cancer stem cells.


• AGNOSTOS trial will establish whether nab-paclitaxel, a more effective taxane analogue, can be safely and

effectively combined to gemcitabine or carboplatin, the two most used CUP drugs, to improve disease control

and outcome of these patients.

• AGNOSTOS translational study will deploy the extensive genomic work-up of enrolled patients to pinpoint

common denominators of the hypermetastatic phenotype and it will exploit patient-derived-xenograft for an

in-depth molecular characterization of these tumors, with the potential to unravel the molecular link between

cancer stemness and the hypermetastatic phenotype.

A forgotten pathology and an unmet theraputic goal

Selected references


Can

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Filippo Montemurro MDUnit for Investigational Clinical Oncology


Fax. +39.011.9933275

Montemurro F, Scaltriti M: Biomarkers of drugs targeting HER-family receptors. J Pathol. 2014, 232:219-229

Montemurro F, et al.Potential markers of long-term benefit from single-agent trastuzumab or lapatinib in HER2-positive metastatic breast cancer. Molecular Oncology. 2014, 8:20-26 Montemurro F, et al. Human epidermal growth factor receptor 2 (HER2)-positive and hormone-receptor positive breast cancer: new insights into molecular interactions and clinical implications. Ann Oncol. 2013, 24:2715-2724 Montemurro F, et al. Hormone receptor expression and activity of trastuzumab with chemotherapy in HER2-positive advanced breast cancer patients. Cancer 2012, 118:17-26


Dr. Filippo Montemurro is the Director of the Investigative Clinical Oncology Unit (INCO) at the Candiolo Cancer

Institute - FPO IRCCS. This recently created Unit is at the intersection between basic, translational, and clinical

research and it is strongly committed to designing clinical trials of cancer precision medicine.

During his fellowship and his professional career, Dr. Montemurro has focused on breast cancer and biologically

targeted therapies for the HER2-positive subset, where he has acquired competence and international visibility.

In particular, he has pioneered reductive approaches aimed at sparing chemotherapy to patients with HER2-

positive metastatic breast cancer whose disease showed molecular features of clinical addiction to the HER2

oncogene.

Currently, he is Principal Investigator and steering committee member of a number of Phase II and III, multi-

centric international Clinical Trials in breast cancer. Furthermore, in the context of the activities of the INCO

Unit, he co-chairs a special program for patients with metastatic Cancer of Unknown Primary origin (CUP),

which includes clinical and molecular investigations in this rare, but meaningful disease subset.

58 | Applied Clinical Research

Modern pathology: from morphology to molecular genetics

Applied Clinical Research | 59

Research Topic 4: Applied Clinical Research

The ultimate task of the Institute is the development of clinical research integrated with health assistance, also through nationally- and internationally-controlled clinical trials. With the introduction of molecular therapies, it has been understood that a specific disease can have different characteristics that need different therapies. Treatments are therefore oriented toward personalized therapy, which needs a very sophisticated molecular diagnostics armamentarium that the Institute is able to provide. Ongoing clinical research integrates the Divisions, the Laboratories and the Facilities, in order to allow synergy among different highly-specialized technologies and complementary, diagnostic, and therapeutic know-how. The research goal is the progress of science, but the figure and the well-being of the patient are an absolute priority. Participation in experimental protocols is voluntary and is suggested to patients according to the biological and clinical characteristics of the disease. The Ethics Committee of IRCCS-FPO guarantees that patients receive only treatments having solid scientific bases and that they are monitored with the most serious attention. In compliance with the Declaration of Helsinki, patients who do not wish to participate in a trial receive the best “standard” therapy.Oncologists, surgeons, radiologists and radiotherapists cooperate with researchers to design clinical trials based on molecular data, intended to verify and generate hypotheses. In order to make this cooperation productive, the Institute manages a daily net of interactions involving formal aspects (seminars and meetings), training (refresher courses and lessons) and operational efforts (contacts with pharmaceutical companies, regulatory and management instruments).Clinical Research is the last and most important haven to improve the assistance to cancer patients, to increase their life expectancy and to provide them with the most appropriate, safe and effective therapeutic approach in accordance with the genetic profile of their own tumor. The controlled clinical Protocol is the final product of the overall work of a multidisciplinary group composed of basic researchers, oncologists, surgeons, pathologists, pharmacologists, radiologists and nuclear medicine doctors, biostatistics and bioinformatics scientists, research healthcare assistants, data managers, and legal experts. The main projects of the Applied Clinical Research are listed below, followed by the table of Controlled Clinical Trials.

Research Topic 4 concerns:4.1. Colorectal cancer: onset and progression4.2. Development of new therapeutic strategies4.3. New approaches to surgical oncology4.4. Laboratory Medicine, Imaging and Radiotherapy


Personalized therapies in Medical OncologyA new avenue of cancer therapy

TeamMarco Fizzotti, Lucia Garetto, Cinzia Ortega, Veronica Prati, Delia Rota Scalabrini, Giorgio Valabrega, Fiorella Ruatta, Celeste Cagnazzo, Caterina Aversa, Rosa La Face, Sofia Genta, Rossella Martinello, Gloria Mittica, Sonia Capellero, Luca Crotto, Marta Fenoglio

Research topic:

Integrating chemotherapy, target therapy and immunotherapy for treatment of advanced cancer.

Background:

The treatment of advanced cancer depends on the combination of chemotherapy, target therapy and recently

added immunotherapy. The knowledge of the tumor genotype and phenotype is essential for the planning of

the treatment at diagnosis. Unfortunately resistance to treatment eventually occurs: its early detection allows

rapid changes of therapy with an improved prognosis. We have set up continuous monitoring of tumor genot-

ype to determine molecular changes induced by the treatment.


Our units have been focusing mainly on the treatment of sarcomas, ovary, genitourinary and digestive tract

carcinomas, becoming a referral center for these diseases. The results achieved derive from a continuous inte-

raction between the laboratory (both the unit’s lab and the other labs of the Institute) and the clinic. This made

possible the implementation of clinical trials designed and conducted on the basis of hypotheses generated at

the bench and translated to the bedside.


To continue our effort to design clinical trials based on a thorough analysis of the tumor and of its interaction

with the host. The secondary projects will detail the ongoing projects coordinated by our Unit. Moreover, strict

cooperation is ongoing with other leading clinical oncological institutions in Italy and in Europe.

Selected references


Can

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Massimo Aglietta MDUnit of Medical Oncology

Full ProfessorUniversity of Torino Medical School


Fax. +39.011.9933299

Grignani G, et al. Sorafenib and everolimus for patients with unresectable high-grade osteosarcoma progressing after standard treatment: a non-randomised phase 2 clinical trial. Lancet Oncology. 2015, 16:98-107

Aglietta M, et al. A phase I dose escalation trial of tremelimumab (CP-675,206) in combination with gemcitabine in chemotherapy-naive patients with metastatic pancreatic cancer. Ann Oncol. 2014, 25:1750-5

Leone F, et al. Panitumumab in combination with infusional oxaliplatin and oral capecitabine for conversion therapy in patients with colon cancer and advanced liver metastases. The MetaPan study. Cancer. 2013, 119:3429-35


Massimo Aglietta started his scientific career working in preclinical models of human tumors with the

aim of developing rational chemotherapeutic schedules based on tumor cell kinetics (Cancer Res, 1979).

A logical pursuit of these studies has been the characterization of the growth pattern of normal and ne-

oplastic hemopoietic progenitor cells (Cancer Res, 1980; Blood, 1986). These preclinical studies led to

the development of phase I/II clinical protocols that contribute to a rational scheduling of hemopoie-

tic growth factors to allow dose dense chemotherapy regimens in solid tumors (J Clin Invest, 1989; Blo-

od, 1993; Cancer, 1993; Eur J Cancer, 1995). This background was also behind the phase II proto-

cols that explored the potential of stem cell transplantation outside hemopoietic malignancies like

colon cancer and sarcomas (J Clin Oncol, 2002; Blood, 2006; Nature Clin Practice Oncol (Review), 2008).

Serving as Director of the Medical Oncology Unit he has continued to pursue the objective of building cli-

nical trials based on strong preclinical backgrounds. Of relevance: 1) the studies on the development

of resistance to anti HER-2 therapy in breast cancer (Review in: Ann Oncol, 2007; Ann Oncol, 2013); 2)

the preclinical studies in bilio-pancreatic cancer that led to the development of phase II clinical trials to

evaluate the antitumor activity of treatments that combine chemotherapy with monoclonal antibodies di-

rected anti EGFr or anti CTL-4 (Ann Oncol, 2014; Review in: Crit Rev Oncol Hematol, 2013; Crit Rev On-

col hematol, 2014) ; 3) Preclinical studies in sarcoma and melanoma revealed molecular alterations

involved in tumor progression. Molecularly driven clinical trials have been successfully conducted (Can-

cer, 2009; Cancer, 2011; Ann Oncol, 2012, Lancet, 2012; Clin Cancer Res, 2013, Lancet Oncol, 2015).

We are now continuing to translate into the clinic the discoveries of basic research (Clin Cancer Res., 2013;

Cancer Res., 2014). We are engaged in clinical trials exploring the best way to integrate immunotherapy with

chemotherapy and molecular based therapy.


Unravelling Triple Negative Breast Cancer (TNBC) diagnosis

TeamLaura Casorzo, Carmine Dell’Aglio, Ivana Sarotto, Elena Frangipane, Alberto Pisacane

To subtype TNBC using a bottom-up approach from cell lines to formalin fixed paraffin embedded (FFPE) tissues

Research Topic:

To subtype TNBC using a bottom-up approach from cell lines to formalin fixed paraffin embedded (FFPE) tissues

Background:

The standard of care of TNBC is still chemotherapy. However, according to gene expression profiling TNBC can

be divided into 6 intrinsic subtypes, which may respond to different therapeutic protocols. In addition, TNBC

show a low frequency of oncogenic driver mutations but a high heterogeneity of gene copy number. This mole-

cular alteration may follow to hypomethylation and activation of Long Interspersed Nuclear Elements (LINE-1).

LINE-1 may reverse transcribe their own mRNAs and insert their DNA copies into new sites within the genome

and in turn impair oncosuppressor genes and activate oncogenes.


We are setting up panels of biomarkers to classify cell lines, profiled for the 6 molecular variants, within each in-

trinsic subtype of TNBC. The clinical significance of these classifiers will be validated in a retrospective cohort

of FFPE TNBC by correlating their expression with patient follow up. In addition, they will serve for annotating

TNBC prospectively collected in order to create a biobank of fresh tissues and organoids. Finally, we are setting

specific procedures to test the methylation status of LINE-1 sequences to explore new mechanisms of chromo-

some and gene instability in TNBC subtypes.


Our study on TNBC may provide specific biomarkers to be used for patient selection in clinical trials. The cell

lines and organoids representing TNBC subtypes will provide key models for preclinical studies with targeted

agents. Epigenetic analysis will shed light on the activation of complex molecular pathway and on the possibility

to suppress the tumor growth.

Selected references


Can

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Anna Sapino MDUnit of PathologyMedical Director


Marchiò C, Maletta F, Annaratone L, Sapino A. The Perfect Pathology Report After Neoadjuvant Therapy. J Natl Cancer Inst Monogr. 2015; 2015:47-50

Okada T, et al. The Rho GTPase Rnd1 suppresses mammary tumorigenesis and EMT by restraining Ras-MAPK signalling. Nat Cell Biol. 2015; 17:81-94.

Balmativola D, et al. Pathological non-response to chemotherapy in a neoadjuvant setting of breast cancer: an inter-institutional study. Breast Cancer Res Treat. 2014; 148:511-23.


Anna Sapino is director of the Division of Pathology. She is full Professor of Pathology at the Medical School

University of Torino, and Professor of the European School of Pathology. She has been Vice-Director for Re-

search of the Dept. of Medical Sciences (2013 - 2015). In 2013 she has been appointed chair of the Working

Group for Breast Pathology of the European Society of Pathology and in 2014 Member of the Board of Gover-

nors of Health (Consiglio Superiore della Sanità) of the Italian Ministry of Health. She started as a diagnostic

pathologist with a vivid curiosity towards breast cancer biology (Cancer Res. 1986). In 1987 she accrued expe-

rience in experimental studies on pre-neoplastic breast lesions through the sabbatical period spent in the USA.

Upon her return to Italy she set-up her own cell biology lab working on effects of hormones on breast cancer

cells and mouse mammary gland development on whole gland organ cultures. These studies provided the first

data suggesting myoepithelial cell involvement in the development of pathological entities occurring in the hu-

man breast (Cancer Res. 1992) and the possible involvement of oxytocin in the proliferation and differentiation

of breast stem cells in myoepithelial cells (Endocrinology. 1993). Currently her scientific activity has two key

missions: (i) to improve the quality of diagnostic tests and innovate breast cancer diagnosis by working on pre-

analytical phase in surgical pathology, and to guarantee the optimal quality for any downstream analysis (PLoS

One. 2011); (ii) to sort out the main drawbacks of oncologists by searching and validating biomarkers that may

guide towards a more personalized therapeutic approach (J Pathol. 2009; J Natl Cancer Inst Monogr. 2015)


Identification of potential targets in cancer of the biliary tractDevelopment of new cancer therapies to improve the prognosis of biliary carcinomas, a group of highly lethal malignances

TeamRenato Ferraris, Federica Colombi, Ilaria Depretis, Elisabetta Fenocchio, Roberto Filippi, Donatella Marino, Giuliana Cavalloni, Caterina Peraldo Neia

Research topic:

New strategies in advanced biliary cancer treatment

Background:

Biliary tract cancer (BTC) is a rare and lethal disease with very few therapeutic options. Previous studies sug-

gest that Epithelial Growth Factor Receptor (EGFR) pathway activation could be involved in BTC pathogenesis,

envisaging a potential role of anti-EGFR monoclonal antibodies.


Despite encouraging early evidence of a major involvement of EGFR pathway in BTC, targeted anti-EGFR therapy

has proven disappointing in the clinic, and resistance seems to involve mechanisms other than RAS mutations.

Our group has coordinated a multicenter randomized Phase II clinical trial (Vecti-BIL study) to evaluate the

effectiveness of Panitumumab (P) in combination with chemotherapy in patients with advanced BTC selected

for the absence of KRAS mutations on exon 2. Patients were randomized to receive the combination of P with

gemcitabine and oxaliplatin (GEMOX) or GEMOX alone. The addition of P did not result in a statistically signifi-

cant improvement in progression-free survival (PFS) (5.3 months vs 4.4 months of GEMOX alone; log-rank test,

p= .27) and had no impact on overall survival (OS, 9.9 months with P-GEMOX vs 10.2 months with GEMOX;

p= .42). Even restricting survival analyses to all RAS and BRAF wild-type patients, no statistically significant

differences in PFS or OS were found between treatment arms.Resistance to chemo/targeted treatments may

be explained through alternative pathway activation or the presence of a stem cell compartment. Comprehen-

sive molecular profiling and genetic analyses of the last 10 years have provided considerable advances in the

identification of druggable targets in BTC. In our preclinical experience, we explored alternative molecules such

as ET-743 and the Src inhibitor Saracatinib. Moreover, we demonstrated that 9% of patients harbor the FIG-

ROS1 rearrangement, in a case series of Italian patients with BTC, suggesting that inhibition of ROS1 may have

important clinical implications.To test new anticancer treatments in BTC we established a human intrahepatic

cholangiocarcinoma cell line derived from an Italian patient and a patient-derived intrahepatic cholangiocarci-

noma xenograft model.


Molecular profiling has outlined complex oncogenic pathway alterations in BTC including ERBB2, RAS-RAF-

MEK, PI3K-AKT-mTOR, JAK/STAT, andIDH1/2. Gene fusions involving ROS1 and FGFR2 represent timely drug-

gable alterations. These findings are expected to provide important implications for developing future therapies

in BTC.

Selected references


Can

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Francesco Leone MDUnit for Investigation of Gastrointestinal Tract Cancer


E-mail: [email protected] Phone. +39.011.9933503 - 3250

Fax. +39.011.9933524/3275

Fornaro L, et al. Second-line chemotherapy in advanced biliary cancer progressed to first-line platinum-gemcitabine combination: a multicenter survey and pooled analysis with published data. J Exp Clin Cancer Res. 2015 Dec 23;34:156.

Peraldo-Neia C, et al. Anti-cancer effect and gene modulation of ET-743 in human biliary tract carcinoma preclinical models. BMC Cancer. 2014 Dec 5;14:918

Cavalloni G, et al. Establishment and characterization of a human intrahepatic cholangiocarcinoma cell line derived from an Italian patient. Tumour Biol. 2015 Oct 20. [Epub ahead of print] PubMed PMID: 26486326.


Francesco Leone is Assistant Professor at the University of Turin, Department of Oncology. Since 2006 he has

been coordinator of the activities of the multidisciplinary team for the treatment of digestive cancers at the

Candiolo Cancer Institute and coordinator of translational research projects in this field between the Medical

Oncology Division and the Oncology Laboratory of the Institute. In colorectal cancer, his major area of interest is

represented by “conversion therapy” to maximize the possibility to offer surgery to metastatic patients (Crit Rev

Oncol Hematol. 2014, Cancer 2013). He realized a significant synergy also for the multidisciplinary treatment

of locally advanced pancreatic cancer (Cancer 2013) and, in the metastatic setting, he participated in the

pioneering of modern immunotherapy concepts (Ann Oncol. 2014). In recent years, his main area of research

has been the study of biliary tumors. He contributed to exploring the feasibility of targeted therapies for these

highly malignant cancers (Crit Rev Oncol Hematol. 2013, Clin Cancer Res. 2006) and is currently engaged in

the final analysis of a multicenter randomized clinical trial of panitumumab in combination with chemotherapy

for advanced biliary tumors, coordinated by the Candiolo Cancer Institute (ASCO GI Proceedings 2015). Given

the unsatisfactory results of the anti-EGFR based strategy, he developed insights in new directions such as the

evaluation of new chemotherapy agents (BMC Cancer 2014) and the identification, through extensive molecular

analysis, of specific genetic targets to be explored in future clinical trials (Genes Chromosomes Cancer 2014).


Cell therapy for metastatic tumorsApplication of new cell-therapy based strategies, to synergize with molecular targeted therapies

TeamDaniela Caravelli, Valentina Coha, Paolo Becco, Susanna Gallo

Research topic:

Hematopoietic Cell Transplantation (HCT): toward an immunological platform for cell and molecular therapy in

the treatment of metastatic refractory cancers.

Background:

Allogeneic HCT, coupling to high dose chemotherapy a powerful donor immunological anti-tumoral effect, is a

curative therapy for many hematological malignancies. However in the past years its broader application has

been limited by two relevant obstacles: the first being represented by Graft Versus Host Disease (GVHD) -the se-

cond being represented by the incapability of generating a specific Graft Versus Tumor effect (GVT). Only when

it will be possible to achieve a complete control of GVHD along with the generation of a specific GVT, allogeneic

HCT will accomplish its extraordinary curative potential. Our research is focused on the clinical application of

a strategy able to fully control GVHD and that transforms HCT into a safe platform for adoptive cell therapy.


In regard to GVHD our group contributed to the study of conventional treatment and classification of this com-

plication and is currently running an interventional study of post-transplant cyclophosphamide (PT-CY) as a

safe strategy for GVHD control. In this context in the first 40 patients treated we reported an extremely low rate

of acute GVHD and of chronic GVHD that translated in a sharp reduction of transplant related mortality (2%).

In regard to GVT (i) we at first revealed that following HCT it was possible to generate donor T cells specifically

directed against patient-tumor antigens. (ii) We contributed to demonstrating that allogeneic HCT is an effective

cell-therapy in the treatment of Multiple Myeloma. (iii) We have been involved in pre-clinical studies regarding

a specific form of cell-therapy derived from the generation and infusion of Cytokine Induced-Killer cells (CIK).

In regard to Cell therapy regulatory issues and Law: cell therapy clinical application will be allowed only in Clini-

cal centers that are JACIE-FACT accredited. Our Transplant center as part of the Turin metropolitan transplant

network was accredited in July 2013.


The introduction in the clinical scenario of a safe strategy for GVHD control, as the on-going results HCT with

PT-CY suggest, represents the ideal basis for future application of tumor-specific T-Cell-therapy. We plan to de-

sign and conduct clinical trials of cell-therapy (CIK and tumor specific T-cells) in the context of the Turin metro-

politan transplant network both in solid tumors (melanomas and sarcomas) and in hematological malignancies.

Selected references


Can

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Fabrizio Carnevale-Schianca MDUnit of Cell Therapy


Fax. +39.011.9933299

Gammaitoni L, et al. Effective Activity of Cytokine Induced Killer Cells against Autologous Metastatic Melanoma including Cells with Stemness Features. Clin Cancer Res. 2014, 19:4347-58

Sanlorenzo M, et al. Synergyof molecular targeted approaches and immunotherapy in melanoma: preclinical basis and clinical perspectives. Expert opinion on biological therapy. 2014, 10:1491-500

Gammaitoni L, et al. Immunotherapy of cancer stem cells in solid tumors: initial findings and future prospective.Expert opinion on biological therapy. 2014, 9:1259-70


Fabrizio Carnevale Schianca has been working in the field of hematopoietic stem cell transplantation (HCT)

since the time of fellowship (1997) accumulating robust clinical knowledge both in the biological rationale and

clinical management of this strategy. As an oncology fellow he contributed to the start-up of the clinical unit

of Medical Oncology at the Candiolo Institute (1998). At the end of his fellowship he spent 18 months at Fred

Hutchinson Cancer Research Center mentored by Rainer Storb. In Seattle he worked in the clinic, gaining deep

experience of allogeneic HCT; in clinical research he has carried out studies on GVHD treatment and classifica-

tion (Biol Blood Marrow Transplant 2000, Bone Marrow Transplant 2009, Biol Blood Marrow Transplant 2009).

Once back in Candiolo he led the opening of the Medical Oncology Stem Cell Transplant Unit (2001); this unit

is now part of a large metropolitan network that was JACIE-FACT accreditad in 2013.

In clinical research, since his return, he has dedicated his efforts to mechanistically demonstrating that allo-

geneic HCT can generate a specific, long lasting graft versus tumor effect in hematological and solid tumors

(Blood 2006, NEJM 2007, Blood 2009, Biol Blood Marrow Transplant 2009, Blood 2011). In the last years his

group efforts have been aimed at exploring strategies for controlling GVHD and transforming HCT into a safe

platform for specific anti-tumoral cell-therapies. In this regard he is collaborating with immunologists (Int Im-

munol 2008, J. immunother 2012). Since 2010 he and his group have been involved in the constitution of a

melanoma-unit (BJC 2014, Cancer Invest, 2014). In metastatic melanoma, taking advantage of his experience

in cellular therapies, he is collaborating with immunologists to project clinical trials aimed at combining targe-

ted therapies and check-point modulators to adoptive cell therapy (Clin Cancer Res 2013, Expert Opinion Biol

Ther 2014).


Integrated therapies for the treatment of sarcomasCooperative trials for new therapeutic approaches

TeamYmera Pignochino, Sandra Aliberti, Lorenzo D’Ambrosio, Danilo Galizia, Federica Capozzi, Paola Boccone, Sara Miano, Erica Palesandro, Marta Canta, Maja Todorovic

Research topic:

Integrated therapies for the treatment of sarcomas

Background:

Bone and soft tissue sarcomas (B-STS) are a rare and heterogeneous group of tumors. As of today, a multidiscipli-

nary treatment encompassing complete surgical removal of the tumor +/- chemotherapy and radiotherapy cures

roughly 50% of patients. Unfortunately, the most active chemotherapies may eradicate micro-metastatic disease,

but do not cure non-resectable disease. Several second- and further-line treatments have been tested showing

marginal activity at most. Therefore, innovative therapeutic strategies are urgently needed.


We focused on the identification of key-pathways that can be exploited with innovative therapeutic tools. In osteo-

sarcoma we identified the role of the mTOR pathway as a mechanism of resistance to sorafenib and showed the

possibility of hitting this pathway with an innovative target combination therapy with sorafenib and everolimus. In

this context we ran and completed a phase II trial. We identified on several B-STS the expression of specific an-

tigens that can elicit a MHC-unrestricted T-cell immune-response. Thereafter, we demonstrated in B-STS patients

that we could sort and ex-vivo expand a specific T-cell sub-population endowed with antitumor activity directed

against patients’ tumors. After ongoing AIFA protocol approval, a phase I/II trial will be run within the Italian Sar-

coma Group.


The achieved results show the importance of dissecting B-STS heterogeneity to identify histotype specific Achilles’

heel. Moreover, we are developing a form of immunotherapy that can be combined with any of the multidisci-

plinary therapies that are the real backbone of sarcoma treatment. In perspective we look for an even greater

integration with the other immunotherapeutic tools directed to reactivating patient anti-tumor response. Secondly,

several new compounds with improved selectivity to targets are in development. We have built a successful part-

nership with several pharmaceutical industries based on clinical trials designed and conducted by our team. On

this ground, we will keep on joining international trials of important drugs, as we did with pazopanib, eribulin, and

investigating combination therapies based on our preclinical/translational research activity.

Selected references


Can

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Giovanni Grignani MDUnit for Investigation and Therapy of Sarcomas


Fax. +39.011.9933299

Sangiolo D, et al., Cytokine-induced killer cells eradicate bone and soft-tissue sarcomas. Cancer Res 2014, 74:119-29

Pignochino Y, et al., The Combination of Sorafenib and Everolimus Abrogates mTORC1 and mTORC2 upregulation in osteosarcoma preclinical models. Clin Cancer Res 2013, 19:2117-31

Stacchiotti S, et al. Phase II study of imatinib in advanced chordoma. J Clin Oncol 2012, 30:914-20

Grignani G, et al. A phase II trial of sorafenib in relapsed and unresectable high-grade osteosarcoma after failure of standard multimodal therapy: an Italian Sarcoma Group study. Ann Oncol 2012, 23:508-16


Giovanni Grignani has been involved in clinical research focusing on the development of prognostic tools in onco-

hematology and later in the field of sarcomas. During his residency he worked on the identification of prognostic

factors to improve multiple myeloma and lymphoma stratification as well as on several phase II trials exploring

innovative chemotherapy combinations. Since he moved to the Candiolo Institute, he has been actively involved

in translational research in the field of stem cells and sarcomas. In particular, he focused on the mechanisms

of metastatic seeding of osteosarcoma (Clin Cancer Res 2005) identifying the CXCR4/CL12 axis as one of the

major determinants of the peculiar pattern of osteosarcoma diffusion. He took part in several pivotal trials of

targeted therapies and took advantage of his large experience to develop preclinical models to test targeted

therapies in sarcomas other than gastrointestinal stromal tumors (Cancer 2011). His group identified the role

of the MAPK pathway (Clin Cancer Res 2013) in osteosarcoma progression and, on this original finding, he

demonstrated the activity of sorafenib and mTOR inhibitor in the advanced stages of this tumor (Ann Oncol

2012 and Lancet Oncol 2015).

In the last years, he set up a preclinical collaboration with immunologists in the attempt to overcome the intrin-

sic limit of target therapies related to tumor heterogeneity. In this setting, he has recently published an original

model to expand patient derived lymphocytes showing that so-called cytokine induced killer cells selectively kill

autologous sarcoma cells (Cancer Res 2014). In the forthcoming year this experience is going to be translated

into the clinic in a phase I/II trial.


Hyperthermic intraperitoneal chemotherapy (HIPEC)Treatment of peritoneal carcinomatosis

Team Armando Cinquegrana, Patrizia Marsanic, Andrea Muratore, Demetro Siatis, Alfredo Mellano, Marco Vaira

Research topic:

Treatment of peritoneal carcinomatosis by surgical cytoreduction combined with HIPEC.

Background:

Our clinical experience is focused on the treatment of peritoneal carcinomatosis and tumor recurrence. Since

1995, about 900 operations for peritoneal carcinomatosis have been performed; since January 2013 to date

more than 150 patients with peritoneal carcinomatosis have been treated in our Istitute. Our operative unit is

specialized in the treatment of this kind of disease by cytoreductive surgery (CRS) combined or not with hyper-

thermic intraperitoneal chemotherapy (HIPEC) and secondary surgical cytoreduction of advanced cancers.


In the last 3 years we have performed about 65 CRS + HIPEC (peritoneal mesothelioma = 11; colorectal carcino-

matosis = 14; pseudomyxoma peritonei = 33; ovarian carcinomatosis = 7). Through the constant improvement

of surgical and perioperative skills, we have achieved promising results in terms of long-term survival (median

overall survival of 65 months for peritoneal mesothelioma, 42 months for ovarian cancer, 58 months for colo-

rectal carcinomatosis, 144 months for PMP associated with a 10-yr overall survival of 80%) associated with

low perioperative morbidity and mortality rates. We have registered an overall major morbidity rate of 15%,

decreased to 6.7% considering only the last three years associated with no perioperative death. Since January

2013 in our Institution we have performed more than 100 surgical cytoreductions, which 65 for ovarian carci-

nomatosis. The mainstay of treatment is the attempt to achieve complete surgical cytoreduction, identified as

the most significant prognostic factor: in this regard we perform a high number of primary or interval debulking

surgeries with no macroscopic residual disease (CC-0) in a substantial number of patients. In cooperation with

medical oncology, considering that the follow-up is too short, the results in terms of long-term outcome are

still being processed. We can assert that in the last two years we registered a major complications rate of 2-3%

with no postoperative deaths. All the HIPEC procedures (65 cases) were carried out intraoperatively with an ori-

ginal “semi-closed” technique. During the last years we have redefined our patient selection policy and sought

to restrict indications to patients with less advanced or less aggressive disease: the selection process for CRS

and HIPEC is critical and prognostic factors are required to identify patients who may most benefit from these

treatments. The standardization of the surgical technique borrowed from experience in the treatment of perito-

neal carcinomatosis allows us to obtain good long-term results with limited costs and mean hospital stay time.


In order to perform a more careful selection of patients, we are investigating the practice of second look (tech-

nique) to identify patients that early present an higher risk of relapse; for that purpose we are, also, testing

the use of the liquid biopsy for the detection of circulating DNA. We are also evalueting laparoscopic HIPEC on

selected patients (PIPAC), but we only have preliminary results.

Selected references


Can

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Michele De Simone MDUnit of Surgical Oncology

E-Mail: [email protected]. +39.011.9933449

Fax. +39.011.9933440

Caneparo A, et al. Contamination risk for operators performing semi-closed HIPEC procedure using cisplatin. Eur J Surg Oncol. 2014, 40:925-9.

Cavaliere F, et al.Prognostic factors and oncologic outcome in 146 patients with colorectal peritoneal carcinomatosis treated with cytoreductive surgery combined with hyperthermic intraperitoneal chemotherapy: Italian multicenter study S.I.T.I.L.O.Eur J Surg Oncol. 2011, 37:148-54.

Baratti D, et al.Multicystic peritoneal mesothelioma: outcomes and patho-biological features in a multi-institutional series treated by cytoreductive surgery and Hyperthermic Intraperitoneal Chemotherapy (HIPEC).Eur J Surg Oncol. 2010 ,36:1047-53.


Michele De Simone has always been interested in research of new technologies in Surgical Oncology. (Antican-

cer Res. 2001). He focused his experience in the field of primary and metastatic tumors taking an active part in

many clinical research projects (Rev Recent Clin Trials 2007). He developed research experience in treatment

of peritoneal carcinomatosis (J Clin Oncol. 2004).


New strategies for breast cancer local-regional controlRe-definition of local-regional breast cancer treatment according to the risk of relapse

TeamFranziska Kubatzki, Furio Maggiorotto, Alessandra Magistris, Francesco Marocco, Stefania Renditore, Alessandro Rivolin, Paola Sgandurra, Salvatore Carlucci

Research topic:

Strategies for breast cancer loco-regional control

Background:

The prevention and early detection of local-regional relapses requires a better understanding of the complex

relationship between the primary tumor and its metastatic dissemination. For optimal breast cancer care new

advancements in surgery and radiotherapy must be integrated in to the complex and rapidly evolving armamen-

tarium of targeted systemic therapies.


The influence of several treatment-related (accelerated partial radiotherapy, width of surgical margins, endocri-

ne therapy with aromatase inhibitors), tumor-related (lobular histology, miR148b expression) and host-related

parameters (body mass index, polymorphisms at the CYP19A1 locus) on the likelihood of breast cancer relapse

have been examined. The available data and the new hypotheses on the relationship between loco-regional con-

trol and survival have been reviewed from a breast surgeon’s standpoint.


The current understanding of breast cancer natural history points towards the existence of a complex interplay

between loco-regional and systemic processes. Loco-regional recurrences are associated with decreased overall

survival and this may be related to a complex relationship between circulating tumor cells, re-seeding of the

primary tumor site and several metabolic effects linked to the act of surgery. A re-definition of local-regional

breast cancer treatment according to the risk of relapse based on improved bio-molecular characterization of

the tumor is required to tailor the adoption of diagnostic tools, loco-regional and systemic treatments.

Selected references


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Riccardo Ponzone MDUnit of Gynecological Oncology


Biglia N, et al. Role of re-excision for positive and close resection margins in patients treated with breast-conserving surgery. Breast. 2014, 23:870-5

Ponzone R, Baum M. Loco-regional therapy and breast cancer survival: searching for a link. Breast. 2013, 22:510-4.

Montemurro F, et al. Omission of axillary dissection after a positive sentinel node dissection may influence adjuvant chemotherapy indications in operable breast cancer patients. Ann Surg Oncol. 2012, 19:3755-61.

D’Alonzo M, et al. Clinical and radiological predictors of nipple-areola complex involvement in breast cancer patients. Eur J Cancer. 2012, 48:2311-8.


In 1987 and 1988, Dr. Ponzone was employed in the laboratories of the Wistar Institute and Children Hospital

of Philadelphia, U.S.A., where he acquired the basics of Molecular Biology and was involved in research projects

on hyperphenylalaninemia and pterin metabolism (J Inher Metab Dis. 1989).

After graduation, he was actively involved in the clinical and research activities at the Dept. of Gynecological

Oncology, University of Turin, Italy, where he specialized in 1997. Over these years, he co-authored several pa-

pers on the prognostic significance of the family of kallikreins in breast and ovarian cancer (Cancer Res 1995).

In 1995 – 1996 he attended as clinical research fellow the Royal Marsden and University College Hospital of

London, UK, where he conducted clinical trails on innovative endocrine therapies for breast cancer (Br J Cancer

1999), and developed a personal interest in the clinical issues related to hereditary breast and ovarian cancer

(Eur J Cancer 1998). From 1999 to 2009 he was appointed attending Gynaecologist at the Mauriziano Hospital

of Turin and at the Institute of Candiolo. During these years he focused his research activity on the adjuvant

(Ann Oncol 2006) and surgical (J Clin Oncol. 2009) therapy of breast cancer. In 2010 he was appointed Director

of the Division of Gynaecological Oncology at the Institute of Candiolo.

His current research interests are devoted to tailoring loco-regional breast cancer treatments (Eur J Cancer.

2012) and minimizing their side effects (Ann Surg Oncol. 2012). He has also consolidated his expertise in pel-

vic surgery, with a particular interest in molecularly-based and innovative treatments of ovarian cancer. In his

career, he has performed more than 4500 breast and pelvic surgeries and has authored more than 90 papers

published in international peer-reviewed journals, as well as book chapters and national guidelines.


Applications of Computer Assisted Diagnosis (CAD) in RadiologyThe diagnostic imaging, improved by CAD systems, allows tumor diagnosis to be brought forward to an early asymptomatic phase

Team Ilaria Bertotto, Delia Campanella, Gabriele Chiara, Andrea Crivellaro, Veronica Deantoni, Valentina Giannini, Antonio Manca, Laura Martincich, Simone Mazzetti, Filippo Russo

Research topic:

Computer Aided Diagnosis (CAD) applications in Radiology

Background:

High resolution images of the human body are generally obtained by Computed Tomography (CT) or Magnetic Re-

sonance (MR) imaging. CT Colonography is an example of such detailed imaging in which data from a CT scanner

are processed by dedicated software to obtain a 3D representation of the colon from inside the intestinal lumen

in a similar way to a conventional endoscope view. On the other side, MR imaging has shown promise in localizing

cancers, because of its intrinsic high soft-tissue resolution, and combining two or more image modalities can im-

prove the sensitivity of MR tests. However the more variables are introduced the more difficult it is to integrate all

available information into one reliable final report, even for an experienced reader. To deal with these complex pro-

blems, computer aided diagnosis systems (CAD) have been introduced to help radiologists in diagnosing disease.


The research group developed a CAD able to produce quantitative maps of the prostate, providing malignancy

probability information with useful data that might be difficult to assess visually from MR scans. Regarding the

CAD colon, diagnostic performance of CT colonography in individuals at increased risk of colorectal cancer was

investigated, and further studies are being conducted using the innovative dual energy technology, to generate so-

called virtual non-contrast images. Finally, within the CAD-breast project, we developed a fully automatic method

to compute the distance between the Nipple-Areola Complex (NAC) and the tumor to preoperatively evaluate the

likelihood of NAC involvement to help select candidates to nipple sparing mastectomy.


Regarding the prostate CAD system, it has been demonstrated useful in automatically detecting prostate cancer

using MR images, showing a per-patient sensitivity of 97% - i.e. at least one PCa lesion was detected in 54 of the

56 patients - and a median number of FP per exam equal to 3 (1st-3rd quartile; 1-4). A multireader study is being

conducted to evaluate if CAD system could help to improve sensitivity and reduce reading time. A new branch of

research is now focusing on assessing the role of CAD in supporting prostatic MR-guided biopsy, and on estima-

ting tumor aggressiveness by integrating the Gleason Score with imaging, in order to better stratify patients and

personalize treatments. Moreover, we are performing a study, whose aim is to compare PCa detection rate of in

bore MR-targeted biopsy with the detection rate of TRUS-guided prostate biopsy in patients with high PSA values

and at least one suspicious region identified by the radiologist at mp-MR imaging.

Selected references


Can

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Daniele Regge MDImage and Data Processing Laboratory (IDP)


Porpiglia F,et al. The roles of multiparametric MRI, PCA3, and PHI: which is the best predictor of prostate cancer after a negative biopsy? Results of a prospective study. J Urol. 2014, pii:S0022-5347:00052-4

Iussich G, et al. CT colonography: preliminary assessment of a double-read paradigm that uses computer-aided detection as the first reader. Radiology. 2013, 268:743-51

Regge D, et al. Efficacy of computer-aided detection as a second reader for 6-9-mm lesions at CT colonography: multicenter prospective trial. Radiology. 2013, 266:168-76


Dr. Regge is the Director of the Radiology Unit at the Candiolo Cancer Institute and he supervises a multidisci-

plinary team, which investigates advanced medical imaging technologies. The group seeks to enhance cancer

diagnosis by means of Computer Aided Diagnosis (CAD) systems. His major scientific accomplishments are

the following: (i) He is internationally recognized for his contributions to the CT-colonography field, being in-

volved in clinical trials and coordinating the largest european multicentre study. This study, together with the

ACRIN-6664, lead to the introduction of CT-colonography into the guidelines of the American Cancer Society

as a screening test for colorectal cancer. (ii) He is PI of the first large world mass screening trial with CT-colo-

nography conducted in Piemonte and in the province of Verona (http://clinicaltrials.gov/show/NCT01739608);

(iii) He is active in the development and clinical validation of CAD systems for diagnosis of colorectal, breast

and prostate cancer. (iv) He has given important contributions to the field of multiparametric-MRI, which helped

change the diagnostic pipeline of prostate cancer. He is the author of more than 100 peer-reviewed publications

and more than 300 scientific contributions in national and international congresses, and he has been mode-

rator or invited speaker at more than 250 conferences worldwide. Dr. Regge has been PI of several supported

research projects, and he is part of the faculty of the European School of Radiology, founding Member and

president elect of the ESOI, board Member and Secretary of the European Society of EuSoMII and President

of the Italian Chapter of Imaging Informatics.


“Personalized” radiation-therapy based on genetic, biological, and “theragnostic” parametersAdapting radiotherapy treatment to the modern concept of “personalized medicine” (“Precision radiotherapy”)

Team Gaetano Belli, Gabriella Cattari, Marco Gatti, Gabriele Petrilli, Antonia Salatino, Antonella Suma, Domenico Gabriele

Research topic:

“Personalized” Radiation-therapy based on genetic, biological, statistical and “theragnostic” parameters.

Background:

The improved IMRT techniques (Intensity Modulated Radiation Therapy) allow the use of high doses, perfectly

“sculpted” on the areas that are going to be treated. A further improvement of radiation treatment is provided

by “daily image control” based on the IGRT technique (Image Guided Radiotherapy). Genomic information and

“theragnostic” data allow the defining models to optimize the use of radiotherapy, in order to personalize tre-

atment.


(i) We have proved - in collaboration with the cancer stem cell laboratory - that ionizing radiation (IR) leads to

the expression of an oncogene (Met) involved in invasion and metastasis control; this data contributes to the

‘vexata questio’ debate about the possible radiotherapy side effects on metastasis sprouting. (ii) We performed

a feasibility study on large fields, using Tomotherapy. (iii) A controlled clinical trial on IMRT-IGRT treatment

protocols for head and neck tumors and recurrences, and three studies on prostate cancers are in progress.

(iv) We are contributing to an international multicenter randomized phase III clinical trial on locally advanced

rectal cancer. (v) We are leading the european project “Computational Horizons in Cancer” to create a predictive

nomogram of prognosis.


We (i) are evaluating the possible impact of genetics and molecular biology on the radiotherapy of Glioblasto-

ma Multiforme (GBM). The preclinical research is implemented by “xenopatients” (immunocompromised mice

transplanted with GBM of single patients, see above); (ii) we are developing personalized radiotherapy treatment

for pancreatic cancer, for clivus cordoma, for pleural mesothelioma and for gynecological tumors; (iii) we are

implementing treatment protocols for microwaves and radiofrequencies and for radiochemo-hyperthermia.

Selected references


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Pietro Gabriele MDUnit of Radiotherapy


Fax. +39.011.9933754

Gatti M, et al. Accelerated partial breast irradiation using 3D conformal radiotherapy: toxicity and cosmetic outcome. Breast. 2013, 22:1136-41.

Valdagni R, et al. Increasing the risk of late rectal bleeding after high-dose radiotherapy for prostate cancer: the case of previous abdominal surgery. Results from a prospective trial.Radiother Oncol. 2012, 103:252-5.

Zucca S, Carau B, Solla I, Garibaldi E, Farace P, Lay G, Meleddu G, Gabriele P. Prostate image-guided radiotherapy by megavolt cone-beam CT. Strahlenther Onkol. 2011, 187:473-8.


Pietro Gabriele is an expert in Radiation Oncology with three medical specializations (Radiology 1979, Onco-

logy 1982 and Otolaryngology 1987); director of Radiotherapy at Mauriziano Hospital (1997), of Sardinia Ra-

dio-Oncology Department (2008) and from 2010 of Radiotherapy in Candiolo. His expertise started with head

and neck cancers, participating in the first Italian phase III trial of Radiochemotherapy for nasopharynx cancer

(JCO, 1988); after that he cooperated in multicenter studies of head and neck cancers (Otol H&N Surg 2000,

Int J Radiat Oncol Biol Phys 2005). He started studies of hyperthermia (1984) publishing results on preclinical

hyperthermia (Cancer Res 2003), hyperthermia alone in recurrent cancers (Tumori, 1989, 75:284 - Cancer

1990), hyperthermia-radiotherapy on parotid cancer (Cancer 1995) and in recurrent breast and melanoma (Int

J Hyperthermia, 2009). In 2011-12 he became board member of the European Society of Hyperthermic On-

cology and in 2014 President of the 29° ESHO congress in Turin. He performed studies on dose escalation in

prostate cancer and published on radiation set-up (Radiother Oncol 2006), IGRT (Stralenther Onkol 2011), and

radiation risk nomograms (Int J Radiat Oncol Biol Phys 2012, Radiother Oncol 2009 and 2012). Since 1999 he

is member of the National Institute of Health board for Quality Assurance in Radiation Oncology (Tumori, 2009,

Radiother Oncol 2007, Crit Rev Oncol-Hematol 2009)


Personalized medicine driven by nuclear molecular imaging in cancer patientsProviding personalized molecular imaging strategies in cancer patients

TeamManuela Racca, Paola Scapoli, Valeria Pirro

Research topic:

Providing Personalized Medicine with Nuclear Molecular Imaging in Cancer Patients - PET/CT in early evalua-

tion of cancer treatment response and targeting resistance to molecular therapies.

Background:

Early differentiation between responders and non responders is a very important issue for the management of

cancer patients. Wide evidence exists that FDG-PET can predict outcomes for many tumor types very soon after

conventional systemic chemotherapy is started. In particular, compared to cytotoxic chemotherapy, many novel

targeted therapies result in disease stabilization rather than tumor shrinkage. The activity of these novel agents

might, therefore, be better reflected by changes in molecular features of the tumor rather than reduction in

size or volume. Recent prospective studies show that a change in tumor glucose activity (measured with FDG

PET standard uptake value-SUV) is a significantly more accurate parameter than a change in size in assessing

histopathological response to neoadjuvant therapy in patient with GIST and high grade soft tissue sarcoma.


The PET imaging facility has been qualified by CoreLab Partners, Inc. for the submission of Nuclear Medicine

and PET images of the patients who are enrolled in the CAUY922A2109 study. The aim of this study is to eva-

luate the efficacy of AUY922 in combination with Trastuzumab in patients with locally advanced or metastatic

HER2-positive breast cancer that has progressed after or during at least one Trastuzumab-containing regimen.


Research goals will be to assess the value of metabolic imaging with PET in predicting response to therapy,

targeting resistance to molecular therapies and improving personalized therapy and prognosis.

Selected references


Can

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Teresio Varetto MDUnit of Nuclear Medicine


Evangelista L., Redana S., Racca M., Geuna E., Vinante L., Zago G., De Carolis V., Ghiotto C., Saladini G., Varetto T. FDG Avidity at PET/CT During Adjuvant Hormonal Therapy in Patients With Breast Cancer. Clinical nuclear medicine. 2014, 39:e135-41.

Varetto T, Durval C. The new UEMS-EACCME criteria for accreditation of live educational events (LEEs): another step forward to improve the quality of continuing medical education (CME) in Europe. European Journal of Nuclear Medicine. 2014, 41:191-6.

Varetto T, Durval C. Continuing Medical Education Committee and UEMS-EACCME. European Journal of Nuclear Medicine. 2013, 40:470-4


Teresio Varetto has more than 25 years’ experience in all diagnostic and therapeutic activities in Nuclear Me-

dicine. During his career he has been a pioneer in the introduction of new diagnostic radiopharmaceutical me-

thodologies in Cardiology (J Am Coll Cardiol, 1993 – Q J Nucl Med Mol Imaging., 2005) and in Oncology (Eur

J Nucl Med Mol Imaging 2005). Since 1998 he is Professor at Turin University Postgraduate Specialization

School in Nuclear Medicine and Postgraduate Specialization School in Hospital Pharmacology. As Senior Expert

in Nuclear Medicine at International Atomic Energy Agency he accomplished Technical Cooperation Expert

Missions in several African Countries collaborating on the creation and dissemination of Nuclear Medicine Units

and giving lectures and counseling at the local University Hospitals.

As chairman of the Continuing Medical Education Committee of the UEMS European Board of Nuclear Medi-

cine he is in charge of accreditation of International educational activities. (Eur J Nucl Med Mol Imaging 2013;

Eur J Nucl Med Mol Imaging 2014; Eur J Nucl Med Mol Imaging 2014). Member of the Executive Board of the

Italian Association of Nuclear Medicine and Molecular Imaging since 2000, he is Chairperson for CME in Nucle-

ar Medicine at the National level. Since 2008, as Director of the Nuclear Medicine Department at the Candiolo

Institute for Cancer Research, he has been involved in research projects aimed at providing personalized me-

dicine to cancer patients as the use of PET/CT in early evaluation of cancer treatment response and targeting

resistance to molecular therapies.

80 | Core Facilities

A mouse ‘xenopatient bearing a human Gliobastoma Multiforme (IVIS scan)

Core Facilities | 81

Core Facilities

inevitably finite, and possibly poor for some tumour subtypes. Therefore, experiments with cell lines cannot recapitulate the wide heterogeneity of human malignancy that occurs among individuals on a population basis. One way to tackle this issue is to perform population-based in vivo studies by using large series of human cancer specimens directly transplanted into mice. This setting can be exploited as a bridgehead to unravel the genetic and biological complexity of cancer, as a prelude for the identification of novel therapeutic targets and unanticipated pharmacological approaches. XPErO provides researchers with a collection of liver metastases from colorectal cancer that have been systematically transplanted in immunocompromised mice to obtain stable tumor lines (xenopatients), which are available for any kind of in vivo/ex vivo study. This resource builds on three main cornerstones: (i) The viable biobank of colorectal cancer liver metastases. An institutional effort has led to the collection and implantation of more than 500 samples of metastatic colorectal carcinoma. This resource allows us to select the appropriate candidates among more than 300 experimental models, based on clinical, biological, genetic or gene expression traits. This, in turn, is instrumental for achieving the appropriate statistical power for any kind of translational study, even if focused on relatively small tumour subpopulations. (ii) The ‘Xenopatients’ experimental platform for in vivo drug screening. We have implemented a dedicated experimental pipeline that required the recruitment of ad-hoc personnel and the development of a specific workflow. The possibility to exploit trained staff and to apply robust standard operating procedures (SOPs) to experimental work confers high added value to research projects, in terms of both cost effectiveness and efficiency of the resources used. (iii) The Laboratory Assistant Suite package for data management and analysis. We have developed a web-based bioinformatics platform that assists biomedical researchers in multiple activities, which range from tracking data generation and SOPs execution to the management of multidimensional molecular profiles and complex data integration (The Laboratory Assistant Suite or LAS, http://devircc.polito.it/wordpress/). This kind of informatic support provides invaluable benefit by optimising quality and reproducibility of the experiments and by accelerating data mining and analysis.

The Oncogenomics Center (OGC)Genetic alterations in tumors are predictors of response or resistance to targeted therapies, and their identification is mandatory for molecular diagnosis and therapeutic decisions. Technological advances in experimental and informatics methodologies over the past 10 years have made possible the characterization of cancer genomes.OGC is the supportive infrastructure for all genomic studies, including transcriptional, mutational and gene copy number analysis of cells, tissues and liquid biopsies. Dedicated personnel and instrumentation are devoted to provide services for qRT-PCR studies, Sanger and Next Generation Sequencing experiments, Gene Expression Array analyses and BEAMing tests.

The ‘Xenopatients’ Platform for Experimental Oncology.Andrea Bertotti, M.D.Assistant ProfessorUniversity of Torino Medical School E-mail: [email protected] Phone. +39.011.9933242

Immortalized cancer cells exhibit a genetic drift, a biological compliance and phenotypic features different from original cancers in patients. Another drawback of the use of continuous cell lines is that the catalogue of currently available models is

82 | Core Facilities

The Bioinformatics Center (BIC)Modern research in cancer biology implies the collection of extensive data from experimental models concerning specific genetic lesions that drive cancer initiation and progression. Such data will include, for example, large sets of expression transcript profiling, comparative genomic hybridization profiling, whole genome sequencing, immunohistochemical data, and morphologic data that will be peculiar to each specific tumor. Thus, a bioinformatics platform for integrated data tracking and normalization is critical to the successful realization of this endeavor. BIC comprises a web-based bioinformatics platform (Laboratory Assistant Suite, LAS; http://devircc.polito.it/wordpress/) that assists biomedical researchers in multiple activities, which range from tracking data generation and execution of standard operating procedures (SOPs) to management of multidimensional molecular profiles and complex data analysis and integration, by managing multiple independent databases that are linked together in an interconnected network (‘oncogrid’).

The Oncology Imaging Center (OIC)Basic research in disciplines such as cell biology, molecular genetics and developmental biology has provided invaluable insights into the regulatory circuits that govern cancer onset and progression. Within this context, we postulate that imaging studies in cell topics and tissues will parallel genomic analyses and in vivo experimentation, constituting an integrative platform for rapid testing of emerging research directions.OIC technologies include comprehensive microscopic imaging systems, such as confocal microscopes, live-cell devices for real-time monitoring of cellular behaviors, and high-throughput platforms for functional screening.

The Flow Cytometry Center (FLOCC)Increasing evidence shows that tumors are structured in a hierarchical form, with a majority of cells undergoing aberrant differentiation but retaining a proliferative capacity limited over time, and a tiny fraction of cancer stem cells (CSCs) or cancer-initiating cells (CICs) that are able to self-renew and continuously regenerate the tumor. FLOCC enables researchers to take advantage of state-of-the-art FACS (fluorescence-activated cell sorter) technologies and dedicated personnel with highly specialized technical skills in order to tackle these issues, by allowing analysis and prospective isolation of individual cancer cells within highly heterogeneous populations.

Clinical Services | 83

Clinical ServicesPiero Fenu, MD Direttore SanitarioE-mail: [email protected] - Phone. +39.011.9933618

The head of the department of clinical services participates in the strategic plan-ning process and contributes, with the formulation of proposals and opinions, to hospital management. He directs health services and is responsible for hygiene and prevention (hospital infection control and environmental hygiene). The clini-cal service office monitors the appropriateness of admissions, the hospital stay lenghts, the average weight of the so called ‘case mix’ and supervises the waiting list. He coordinates with the Operation Department the proper use of spaces and the timely flow of goods and services. The clinical service office:(i) is in charge of the control of the fulfillement of results obtained within the as-signed objectives; (ii) coordinates the facilities of the hospital including the provi-sion of appropriate protocols; (iii) defines strategies and guidelines for extraordi-nary and/or urgent intervention; (iv) advises on building planning regarding further hospital expansion; (v) supervises the ‘conventional’ agreements and the related relations with the University.

Felicino De Bernardi, MD Cancer PainE-Mail: [email protected] - Phone.+39.011.9933018

Cancer pain significantly affects the physical, psychological and emotional com-ponents and quality of life of the individual. Our goal is the optimisation of pain relief through a patient-centred approach with particular attention to quality of life, personalized care and application of the newest drugs and treatments.

Antonino Sottile, MDDirettore Operativo and Laboratory MedicineE-Mail: [email protected] - Phone +39.011.9933038

This Unit is mainly devoted to patient diagnosis and treatment. Nevertheless, a strong effort is made in performing analysis supporting clinical trials as well as translational research. The laboratory is also engaged in its own research activity aimed at developing and testing novel diagnostic procedures. A special emphasis is placed on the analysis of prostate cancer markers.

Giovanni Succo, MDOtolaryngologyE-mail: @ircc.it - Phone. +39.011.9933663

The Otolaryngology Unit has many related clinical functions focused on with the diagnosis and treatment of disorders and diseases of the head and neck. These specifically includes illness or injury of the ear, nose, and throat, as well as medi-cal or surgical treatments for diseases of the skull base.

84 | Clinical Services

Giovanni Galatola, MD Gastroenterology E-Mail: [email protected] - Phone +39.011.9933410

The Gastroenterology Unit at IRCC is a dedicated provider of a wide range of dia-gnostic and therapeutic endoscopic procedures for the upper and lower gastroin-testinal tracts, and is involved in the clinical care of patients with chronic viral hepatitis and those with gastrointestinal disorders in the medical oncology setting.

Alessandro Bonzano, MD CardiologyE-Mail: [email protected] - Phone +39.011.993.3888

The Cardiology Unit at the IRCCS is engaged in the prevention, diagnosis and treatment of cardiotoxicity caused by old chemotherapy drugs (anthracyclines, 5-fluorouracil) and new agents (trastuzumab). Doppler echocardiography and bio-markers as troponin are used to identify early damage of the heart during che-motherapy. A growing experience in the field of Cardioncology has lead to tailored treatment for cancer patients with cardiovascular complications.

Franca Goffredo, MD Pharmacy E-Mail: [email protected] - Phone +39.011.9933261

The Pharmacy Unit manages drug/medication related activities and issues for all IRCCS wards (drug lifecycle). Some of these activities include supplying, storage, distribution, drug information, therapeutic policy/formulary. Preparation of chemo-therapy and supportive care medications for in and out patients, together with in-vestigational drug management and research support (including regulatory issues) are the main and distinguishing activities, enhanced by integrated IT and automa-tion. The staff is an active partner of the University in the pharmacy education and training programs.

Michele Stasi, M.D.Medical PhysicsE-mail: [email protected] - Phone. +39.011.9933708

The Medical Physics Unit is focused on patients’ tumor segmentation based on multi-modal imaging, sophisticated patient specific Quality Assurance and tre-atment options such us Helical Tomotherapy (HT), Intensity Modulated Radiation Therapy (IMRT), Image Guided Radiation Therapy (IGRT), and Adaptive Radiothe-rapy. The use of sophisticated imaging tools (MRI, PET/CT) and IMRT, IGRT may help to achieve better conformal radiation dose to target with respect to 3D stan-dard conformal radiotherapy

Clinical Services | 85

2015 IRCC International Conference on Cancer Precision Medicine

86 | Educational Activities

Educational Activities | 87

Educational Activities: the Chiron international program

IRCC Cancer Conferences• “From Signal Transduction to Cancer Precision Medicine” - June 6, 2015• “International conference on Cancers of Unknown Primary” - May 20-21, 2017

2015 Seminars• Regulation of self renewal in Cancer Stem Cells - Prof. Pier Giuseppe Pelicci, December 11, 2015• p140Cap is a Chromosome 17q12-q21 scaffold protein that limits ErbB2 breast cancer progression - Prof.ssa Paola Defilippi, December 4, 2015• Every Cell Has A Story - Advances in Single Cell Genomics - Dr. Amy Hamilton, December 2, 2015• NanoBiT Complementation Reporter Provides Accurate Assessment of Intracellular Protein Interactions under Physiological Conditions - Keith V. Wood, PhD, November 9, 2015• Data intensive biology and data provenance graphs, ovvero l’importanza di tracciare i dati - Prof. Gianluigi Zanetti, October 12, 2015• Therapeutic strategies based on cancer stem cell targeting . Prof. Ruggero De Maria, October 2, 2015• The Network Genomic Medicine: A Platform to implement Personalised Cancer Treatment in Germany - Prof. Reinhard Büttner, October 1, 2015• Genomics and Therapeutic Opportunities in Gastroesophageal Cancer - Dr. Adam Bass, October 1, 2015• Neuropilin and Integrin Control of Cancer Stem Cell Fate - Dr. Arthur M.Mercurio, Friday, March 20, 2015• NK cells: from surface receptors to the cure of high risk leukemias - Prof. Lorenzo Moretta, March 19, 2015• Colon cancer stem cell resistance to therapy - Dr. Jan Paul Medema, January 26, 2015

PhD ProgramsThe Institute offers four different PhD programs.(a) PhD in Molecular Medicine (Cell Sciences and Technology): a four-year course (under the auspices of the University of Torino) open to young graduates in Medicine, Biological Sciences, Biotechnology, Chemistry, and Pharmacology. It is aimed at training for basic and translational research in biomedical sciences.(b) PhD in Complex Systems and Post Genomic Biology: a three-year course (under the auspices of the Univer-sity of Torino) aimed at training graduates in the fields at the interface of medicine, life sciences, mathematics and physics.(c) PhD Program in Biomedical Sciences and Oncology: a four-year course (under the auspices of the Univer-sity of Torino) open to graduates in Medicine, and Biological Sciences and mainly aimed at training clinical researchers.(d) PhD program in Oncological Sciences: a three-year course (under the auspices of the Catholic University of Milano-Roma) for exceptionally talented graduate students to be trained in cutting-edge oncological research at the highest level.

The Institute provides teaching and training programs in the area of basic and clinical cancer research.These are listed in order of increasing complexity as follows: (a) Courses. (b) Seminars and Workshops. (i) “Lezioni Magistrali”: one in basic research and one in clinical rese-arch, each held by eminent scientists. (ii) Formal seminars: throughout the year, divided among ba-sic and clinical research, held by external invited speakers. (iii) Progress reports held by mem-bers of the Institute, once a week for 48 weeks of the year. (iv) International Workshops, on either basic or clinical research. These workshops are usually organised within the framework program of the Eu-ropean Molecular Biology Organization (EMBO) and of the European Society for Medical Oncology (ESMO). (c) Research Doctorates focused on basic and clinical research training. (d) Master in Molecular Oncology. (e) Post-doctoral programs.

88 | Educational Activities

Graduate students are provided with an individual tailored guidance program and required to rotate through a number of laboratories to learn different research techniques collaborating with world-class scientists. Career counselors help students to investigate job types, develop curriculum vitae and resumes, and refine interview skills. After completing the PhD program students are encouraged to participate in internship opportunities in both basic or clinical research, technology transfer, science policy, and management.All programs are open to Italian and foreign students. Fellowships are provided on a competitive basis

Post-doctoral ProgramsIRCC International Cancer Research Training ProgramThis scheme employs scientists of all nationalities to improve scientific exchange between the Institute’s resear-chers and colleagues who have matured qualifying experience in foreign institutions. The research field is at in the interface between molecular biology and medicine, referring in particular to today’s problems in molecular oncology.

The Marie Curie European Training CenterThe European Commission has recognized IRCC as a qualified center for training young researchers in the experimental medical science field. Within the framework of the program, IRCC employs in rotation graduate students from member states for periods varying from six months to two years in the research laboratories.

Molecular Oncology MasterThe Institute, under an agreement with the University of Torino, also offers a two-year Master in Molecular Onco-logy aimed at providing clinical oncologists, surgical oncologists, radiologists, radiotherapists, and pathologists with up-to-date knowledge of cancer biology and genetics as well as of the novel diagnostic and therapeutic (e.g. precision medicine) approaches that arose from latest molecular and genomic information.

Grant Office and Research Administration - EDP | 89

The Scientific Director’s Office supervises, coordina-

tes and manages the basic, translational and clinical

research at the Institute. To this end, the Grant Office

provides the scientific support for Institute research ac-

tivities and acts as a liaison between researchers and

funding agencies.

The Grant Office assists researchers in identifying ap-

propriate research funding opportunities, centralizing

all information on major national and international, pri-

vate or public, agencies, foundations, and institutions

that support research. Whenever a call for a research

grant is issued, it is advertised throughout the Institute

by e-mail. The Grant Office provides assistance to rese-

archers throughout the application process, in drawing

up budgets - together with the administrative office of

the Institute -, in interpreting the regulations of the

granting agencies, assuring compliance with the spon-

sors’ policies and requirements. In case of successful

outcome of the proposals, the Grant Office helps in pre-

paring reports, consortium agreements with collabora-

ting institutions, and renewal forms.

The Office of Research Administration (ORA) handles

the administration of research funds, assigned to the

Fondazione del Piemonte per l’Oncologia (FPO) and to

the Fondazione Piemontese per la Ricerca sul Cancro

(FPRC). The staff consists of 5 people.

The office manages the registration of every accounting

movement:

i) organization of the first cash note; ii) revenues and

payments; iii) management of personnel; iv) drafting of

the research budget.

With the support of the Grants Office, ORA prepares

economic reports on the costs incurred, in accordance

with the requirements of the single funding agencies,

providing the required supporting documentation.

In 2015, the Office has managed 4347 invoices (2521

by the FPO administration, 1826 by the FPRC admini-

stration), has issued 3025 orders (1587/1438) and has

drawn 125 employment contracts (94/31).

Scientific Management, Grant Office and Research Administration

Daniela Gramaglia PhDE-mail: [email protected]

Phone: +39.011.9933211

Michelina Bruno MsE-mail: [email protected]

Phone: + 39.011.9933237

Antonella Cignetto MsE-mail: [email protected]

Phone: +39.011.9933601

90 | List of Publications

The EDP unit develops and maintains the Master Data-

base, a unique tool, developed in-house, which tracks

the funds, human resources and research publication

output, of the Research Units at the Institute. The EDP

unit runs the Candiolo Cancer Institute public web site

and the Intranet. The unit also provides IT support to

the Research departments and to the Institute’s mee-

tings and conferences.

Electronic Data Processing Unit

Damion Milne E-mail: [email protected]

Phone: +39.011.9933985

Ethics Committee

Luisa Gioeni PhDE-mail: [email protected]: +39.011.9933959

The ethics committee, following the European Directive

2001/20/EC, is an independent body of the Candiolo

Cancer Institute, consisting of healthcare professionals

and non-medical members, whose responsibility is to pro-

tect the rights, safety and well being of human subjects

involved in a clinical trial and to provide public assurance

of that protection, by, among other things, expressing an

opinion on the clinical trial protocol, the suitability of the

investigators involved in the trial and the adequacy of fa-

cilities, and on the methods and documents to be used to

inform trial subjects and obtain their informed consent.

List of Publications | 91


Alajati A, Guccini I, Pinton S, Garcia-Escudero R, Bernasocchi T, Sarti M, Montani E, Rinaldi A, Montemurro F,

Catapano C, Bertoni F, Alimonti A.Interaction of CDCP1 with HER2 enhances HER2-driven tumorigenesis and

promotes trastuzumab resistance in breast cancer. Cell Rep, 2015,11:564-76 - IF: 8,358

Alamo P, Gallardo A, Di Nicolantonio F, Pavan MA, Casanova I, Trias M, Mangues MA, Lopez-Pousa A, Villaverde

A, Vazquez E, Bardelli A, Cacspedes MV, Mangues R.Higher metastatic efficiency of KRas G12V than KRas G13D

in a colorectal cancer model.The FASEB journal, 2015,29:464-76 - IF: 5,043

Alizadeh AA, Aranda V, Bardelli A, Blanpain C, Bock C, Borowski C, Caldas C, Califano A, Doherty M, Elsner M,

Esteller M, Fitzgerald R, Korbel JO, Lichter P, Mason CE, Navin N, Pe’er D, Polyak K, Roberts CW, Siu L, Snyder A,

Stower H, Swanton C, Verhaak RG, Zenklusen JC, Zuber J, Zucman-Rossi J.Toward understanding and exploiting

tumor heterogeneity. Nature medicine, 2015, 21:846-53 - IF: 27,363

Amirouchene-Angelozzi N, Schoumacher M, Stern MH, Cassoux N, Desjardins L, Piperno-Neumann S, Lantz O,

Roman-Roman S.Upcoming translational challenges for uveal melanoma.

British journal of cancer, 2015, 113:1249-53 - IF: 4,836

Arena S, Bellosillo B, Siravegna G, Martínez A, Cañadas I, Lazzari L, Ferruz N, Russo M, Misale S, González I,

Iglesias M, Gavilan E, Corti G, Hobor S, Crisafulli G, Salido M, Sánchez J, Dalmases A, Bellmunt J, De Fabritiis

G, Rovira A, Di Nicolantonio F, Albanell J, Bardelli A, Montagut C.Emergence of Multiple EGFR Extracellular Mu-

tations during Cetuximab Treatment in Colorectal Cancer.

Clinical cancer research, 2015, 21:2157-66 - IF: 8,722

Arpino G, Michelotti A, Truini M, Montemurro F, Russo S, Palumbo R, Zamagni C, Latorre A, Bruzzese D, Riccardi

F, De Laurentiis M, Beano A, Biganzoli L, Zaniboni A, Laudadio L, Malagoli M, Bilancia D, Schettini F, Giuliano M,

Cazzaniga ME, De Placido S.Demographic, tumor and clinical features of clinical trials versus clinical practice

patients with HER2-positive early breast cancer: results of a prospective study.

Journal of cancer research and clinical oncology, 2015, 142:669-78 - IF: 3,081

Assi E, Cervia D, Bizzozero L, Capobianco A, Pambianco S, Morisi F, De Palma C, Moscheni C, Pellegrino P,

Clementi E, Perrotta C.Modulation of Acid Sphingomyelinase in Melanoma Reprogrammes the Tumour Immune

Microenvironment. Mediators of inflammation, 2015, 2015:370482 - IF: 3,236

Barault L, Amatu A, Bleeker FE, Moutinho C, Falcomatà C, Fiano V, Cassingena A, Siravegna G, Milione M, Cas-

soni P, De Braud F, Rudà R, Soffietti R, Venesio T, Bardelli A, Wesseling P, de Witt Hamer P, Pietrantonio F, Siena

S, Esteller M, Sartore-Bianchi A, Di Nicolantonio F.Digital PCR quantification of MGMT methylation refines

prediction of clinical benefit from alkylating agents in glioblastoma and metastatic colorectal cancer.Annals of

oncology, 2015, 26:1994-9 - IF: 7,04

Barrow TM, Barault L, Ellsworth RE, Harris HR, Binder AM, Valente AL, Shriver CD, Michels KB.Aberrant methy-

lation of imprinted genes is associated with negative hormone receptor status in invasive breast cancer.Interna-

tional journal of cancer. Journal international du cancer, 2015, 137:537-47 - IF: 5,085

Bartolini A, Di Paolo D, Noghero A, Murgia D, Sementa AR, Cilli M, Pasqualini R, Arap W, Bussolino F, Ponzoni

M, Pastorino F, Marchio S.The neuronal pentraxin-2 pathway is an unrecognized target in human neuroblastoma

which also offers prognostic value in patients.Cancer research, 2015, 75:4265-71 - IF: 9,329


Bertotti A, Papp E, Jones S, Adleff V, Anagnostou V, Lupo B, Sausen M, Phallen J, Hruban CA, Tokheim C,

Niknafs N, Nesselbush M, Lytle K, Sassi F, Cottino F, Migliardi G, Zanella ER, Ribero D, Russolillo N, Mellano A,

Muratore A, Paraluppi G, Salizzoni M, Marsoni S, Kragh M, Lantto J, Cassingena A, Li QK, Karchin R, Scharpf

R, Sartore-Bianchi A, Siena S, Diaz LA, Trusolino L, Velculescu VE.The genomic landscape of response to EGFR

blockade in colorectal cancer. Nature, 2015, 526:263-7 - IF: 41,456

Bertotti A, Sassi F.Molecular Pathways: Sensitivity and Resistance to Anti-EGFR Antibodies.


Bigatto V, De Bacco F, Casanova E, Reato G, Lanzetti L, Isella C, Sarotto I, Comoglio PM, Boccaccio C.TNF-a

promotes invasive growth through the MET signaling pathway. Molecular oncology, 2015, 9:377-88 - IF: 5,331

Bisaro B, Sciortino M, Colombo S, Camacho-Leal MP, Costamagna A, Castellano I, Montemurro F, Rossi V, Va-

labrega G, Turco E, Defilippi P, Cabodi S.p130Cas scaffold protein regulates ErbB2 stability by altering breast

cancer cell sensitivity to autophagy. Oncotarget, 2015, doi: 10.18632/oncotarget.6710: - IF: 6,359

Caffo O, De Giorgi U, Fratino L, Alesini D, Zagonel V, Facchini G, Gasparro D, Ortega C, Tucci M, Verderame F,

Campadelli E, Lo Re G, Procopio G, Sabbatini R, Donini M, Morelli F, Sartori D, Zucali P, Carrozza F, D’Angelo A,

Vicario G, Massari F, Santini D, Sava T, Messina C, Fornarini G, La Torre L, Ricotta R, Aieta M, Mucciarini C, Zu-

stovich F, Macrini S, Burgio SL, Santarossa S, D’Aniello C, Basso U, Tarasconi S, Cortesi E, Buttigliero C, Ruatta

F, Veccia A, Conteduca V, Maines F, Galligioni E.Clinical Outcomes of Castration-resistant Prostate Cancer Tre-

atments Administered as Third or Fourth Line Following Failure of Docetaxel and Other Second-line Treatment:

Results of an Italian Multicentre Study. European urology, 2015, 68:147-53 - IF: 13,938

Cantini L, Medico E, Fortunato S, Caselle M.Detection of gene communities in multi-networks reveals cancer

drivers.Scientific reports, 2015, 5:17386 - IF: 5,578

Cargnelutti M, Corso S, Pergolizzi M, Mévellec L, Aisner DL, Dziadziuszko R, Varella-Garcia M, Comoglio PM,

Doebele RC, Vialard J, Giordano S.Activation of RAS family members confers resistance to ROS1 targeting

drugs. Oncotarget, 2015, 6:5182-94 - IF: 6,359

Carmona FJ, Montemurro F, Kannan S, Rossi V, Verma C, Baselga J, Scaltriti M.AKT signaling in ERBB2-ampli-

fied breast cancer. Pharmacology & therapeutics, 2015, 158:63-70 - IF: 9,723

Cavalloni G, Peraldo-Neia C, Sassi F, Chiorino G, Sarotto I, Aglietta M, Leone F.Establishment of a patient-derived

intrahepatic cholangiocarcinoma xenograft model with KRAS mutation. BMC cancer, 2015, 16:90 - IF: 3,362

Cavalloni G, Peraldo-Neia C, Varamo C, Casorzo L, Dell’Aglio C, Bernabei P, Chiorino G, Aglietta M, Leone

F.Establishment and characterization of a human intrahepatic cholangiocarcinoma cell line derived from an

Italian patient.Tumor Biology, 2015, doi: 10.1007/s13277-015-4215-3: - IF: 3,611

Celina GG, Rivas MA, Ibrahim YH, Calvo MT, Gris-Oliver A, Rodriguez O, Grueso J, Anton P, Guzman M, Aura C,

Nuciforo P, Jessen K, Argiles G, Dienstmann R, Bertotti A, Trusolino L, Matito J, Vivancos A, Chicote I, Palmer

HG, Tabernero J, Scaltriti M, Baselga J, Serra V.MEK plus PI3K/mTORC1/2 therapeutic efficacy is impacted by

TP53 mutation in preclinical models of colorectal cancer.

Clinical cancer research : an official journal of the American Association for Cancer Research, 2015, 21:5499-

510 - IF: 8,722


Conti A, Majorini MT, Elliott R, Ashworth A, Lord CJ, Cancelliere C, Bardelli A, Seneci P, Walczak H, Delia D, Lecis

D.Oncogenic KRAS sensitizes premalignant, but not malignant cells, to Noxa-dependent apoptosis through the

activation of the MEK/ERK pathway. Oncotarget, 2015, 6:10994-1008 - IF: 6,359

Cossu I, Bottoni G, Loi M, Emionite L, Bartolini A, Di Paolo D, Brignole C, Piaggio F, Perri P, Sacchi A, Curnis

F, Gagliani MC, Bruno S, Marini C, Gori A, Longhi R, Murgia D, Sementa AR, Cilli M, Tacchetti C, Corti A, Sam-

buceti G, Marchiò S, Ponzoni M, Pastorino F.Neuroblastoma-targeted nanocarriers improve drug delivery and

penetration, delay tumor growth and abrogate metastatic diffusion. Biomaterials, 2015, 68:89-99 - IF: 8,557

Del Mastro L, De Placido S, Bruzzi P, De Laurentiis M, Boni C, Cavazzini G, Durando A, Turletti A, Nisticò C, Valle

E, Garrone O, Puglisi F, Montemurro F, Barni S, Ardizzoni A, Gamucci T, Colantuoni G, Giuliano M, Gravina A,

Papaldo P, Bighin C, Bisagni G, Forestieri V, Cognetti F,.Fluorouracil and dose-dense chemotherapy in adjuvant

treatment of patients with early-stage breast cancer: an open-label, 2 × 2 factorial, randomised phase 3 trial.

Lancet, 2015, 385:1863-72 - IF: 45,217

Di Blasio L, Gagliardi PA, Puliafito A, Sessa R, Seano G, Bussolino F, Primo L.PDK1 regulates focal adhesion

disassembly by modulating endocytosis of 3 integrin. Journal of cell science, 2015, 128:863-77 - IF: 5,432

Di Filippo F, Giannarelli D, Bouteille C, Bernet L, Cano R, Cunnick G, Sapino A.Elaboration of a nomogram to

predict non sentinel node status in breast cancer patients with positive sentinel node, intra-operatively assessed

with one step nucleic acid amplification method. Journal of Experimental & Clinical Cancer Research, 2015,

34:136 - IF: 4,429

Elia AR, Circosta P, Sangiolo D, Bonini C, Gammaitoni L, Mastaglio S, Genovese P, Geuna M, Avolio F, Inghirami

G, Tarella C, Cignetti A.Cytokine-induced killer cells engineered with exogenous T-cell receptors directed against

melanoma antigens: enhanced efficacy of effector cells endowed with a double mechanism of tumor recogni-

tion. Human gene therapy, 2015, 26:220-31 - IF: 3,755

Errichiello E, Balsamo A, Cerni M, Venesio T.Mitochondrial variants in MT-CO2 and D-loop instability are invol-

ved in MUTYH-associated polyposis.Journal of molecular medicine (Berlin, Germany), 2015, 93:1271-81 - IF:

5,107

Erriquez J, Becco P, Olivero M, Ponzone R, Maggiorotto F, Ferrero A, Scalzo MS, Canuto EM, Sapino A, Verdun di

Cantogno L, Bruna P, Aglietta M, Di Renzo MF, Valabrega G.TOP2A gene copy gain predicts response of epithe-

lial ovarian cancers to pegylated liposomal doxorubicin: TOP2A as marker of response to PLD in ovarian cancer.

Gynecologic oncology, 2015, 138:627-33 - IF: 3,774

Fornaro L, Vivaldi C, Cereda S, Leone F, Aprile G, Lonardi S, Silvestris N, Santini D, Milella M, Caparello C, Mu-

settini G, Pasquini G, Falcone A, Brandi G, Sperduti I, Vasile E,.Second-line chemotherapy in advanced biliary

cancer progressed to first-line platinum-gemcitabine combination: a multicenter survey and pooled analysis

with published data. Journal of Experimental & Clinical Cancer Research, 2015, 34:156 - IF: 4,429

Frau C, Loi R, Petrelli A, Perra A, Menegon S, Kowalik MA, Pinna S, Leoni VP, Fornari F, Gramantieri L, Ledda-Co-

lumbano GM, Giordano S, Columbano A.Local hypothyroidism favors the progression of preneoplastic lesions

to hepatocellular carcinoma in rats. Hepatology, 2015, 61:249-59 - IF: 11,055


Gagliardi PA, di Blasio L, Primo L.PDK1: A signaling hub for cell migration and tumor invasion.

Biochimica et biophysica acta, 2015, 1856:178-188 - IF: 7,845

Gagliardi PA, Puliafito A, di Blasio L, Chianale F, Somale D, Seano G, Bussolino F, Primo L.Real-time monitoring

of cell protrusion dynamics by impedance responses. Scientific reports, 2015, 5:10206 - IF: 5,578

Garassino MC, Marsoni S.A lesson from vorinostat in pleural mesothelioma.

The lancet oncology, 2015, 16:359-60 - IF: 24,725

Germano A, Rapa I, Volante M, De Francia S, Migliore C, Berruti A, Papotti M, Terzolo M.RRM1 modulates mito-

tane activity in adrenal cancer cells interfering with its metabolization.

Mol Cell Endocrinol, 2015, 5:105-10 - IF: 4,405

Geuna E, Milani A, Martinello R, Aversa C, Valabrega G, Scaltriti M, Montemurro F.Buparlisib , an oral pan-PI3K

inhibitor for the treatment of breast cancer.

Expert opinion on investigational drugs, 2015, 24:421-31 - IF: 5,528

Geuna E, Roda D, Rafii S, Jimenez B, Capelan M, Rihawi K, Montemurro F, Yap TA, Kaye SB, De Bono JS, Molife

LR, Banerji U.Complications of hyperglycaemia with PI3K-AKT-mTOR inhibitors in patients with advanced solid

tumours on Phase I clinical trials. British journal of cancer, 2015, 113:1541-7 - IF: 4,836

Grignani G, Palmerini E, Ferraresi V, D’Ambrosio L, Bertulli R, Asaftei SD, Tamburini A, Pignochino Y, Sangiolo D,

Marchesi E, Capozzi F, Biagini R, Gambarotti M, Fagioli F, Casali PG, Picci P, Ferrari S, Aglietta M; for the Italian

Sarcoma Group.Sorafenib and everolimus for patients with unresectable high-grade osteosarcoma progressing

after standard treatment: a non-randomised phase 2 clinical trial.

The lancet oncology, 2015, 16:98-107 - IF: 24,725

Hultberg A, Morello V, Huyghe L, De Jonge N, Blanchetot C, Hanssens V, de Boeck G, Silence K, Festjens E,

Heukers R, Roux B, Lamballe F, Ginestier C, Charafe-Jauffret E, Maina F, Brouckaert P, Saunders M, Thibault A,

Dreier T, De Haard H, Michieli P.Depleting MET-expressing tumor cells by ADCC provides a therapeutic advanta-

ge over inhibiting HGF/MET signaling. Cancer research, 2015, 75:3373-83 - IF: 9,329

Iacovelli R, Farcomeni A, Sternberg CN, Carteni’ G, Milella M, Santoni M, Cerbone L, Di Lorenzo G, Verzoni E,

Ortega C, Sabbatini R, Ricotta R, Messina C, Lorusso V, Atzori F, De Vincenzo F, Sacco C, Boccardo F, Valduga

F, Massari F, Baldazzi V, Cinieri S, Mosca A, Ruggeri EM, Berruti A, Procopio G.Prognostic factors in patients

receiving third-line targeted therapy for metastatic renal cell carcinoma.

The Journal of Urology, 2015, 193:1905-10 - IF: 4,471

Isella C, Terrasi A, Bellomo SE, Petti C, Galatola G, Muratore A, Mellano A, Senetta R, Cassenti A, Sonetto C,

Inghirami G, Trusolino L, Fekete Z, De Ridder M, Cassoni P, Storme G, Bertotti A, Medico E.Stromal contribution

to the colorectal cancer transcriptome. Nature Genetics, 2015, 47:312-9 - IF: 29,352

Kavuri SM, Jain N, Galimi F, Cottino F, Leto SM, Migliardi G, Searleman AC, Shen W, Monsey J, Trusolino L, Ja-

cobs SA, Bertotti A, Bose R.HER2 Activating Mutations Are Targets for Colorectal Cancer Treatment.

Cancer discovery, 2015, 5:832-41 - IF: 19,453


Kowalik MA, Sulas P, Ledda-Columbano GM, Giordano S, Columbano A, Perra A.Cytokeratin-19 positivity is ac-

quired along cancer progression and does not predict cell origin in rat hepatocarcinogenesis.

Oncotarget, 2015, 6:38749-63 - IF: 6,359

Kwak EL, Ahronian LG, Siravegna G, Mussolin B, Godfrey JT, Clark JW, Blaszkowsky LS, Ryan DP, Lennerz JK,

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