Paradigm shift in solid cancer and the need for antimetastatic drugs · Affordable cancer care....

Paradigm shift in solid cancer and

the need for antimetastatic drugs

Assoc. Prof. Jan Brábek, Ph.D.

Charles University, Faculty of Science

Deparment of Cell Biology and BIOCEV

Jan Brábek, Charles University and BIOCEV 1st April 2020

In 2019, the problems with solid cancer are:

ineffective, unsafe, and unaffordable

medicines!

Do cancer drugs improve survival or quality of life?

You don’t need to know, according to our broken regulatory system

Vinay Prasad

• between 2008 and 2013 both FDA and EMA mostly approved cancer drugs without evidence of

prolonged survival or improved quality of life

• less than 15% were shown later to improve survival

• the median improvement in survival among patients treated with 71 drugs for solid cancer was

just 9 weeks!

Tab. 1. Mortality rates (years 1950-2008) for infectious, heart, cerebrovascular diseases (From

American Cancer Society (ACS) 2010 Cancer Facts & Figures; Atlanta, USA, 2008, modified).

Estimation of the Percentage of US Patients With Cancer Who Benefit

From Genome-Driven Oncology

John Marquart, BA; Emerson Y. Chen, MD; Vinay Prasad, MD, MPH

Question How many US patients with cancer benefit from genome-targeted therapies?

Findings

• in 2006 - 5% of patients were eligible, in 2018 - 8%

• In 2006 - less than 1% of patients responded, in 2018 - 5%



medicines!

It seems as if we are now entering into the WE KNEW IT

ALL ALONG phase of the cancer paradigm shift. Not long

ago, the author of the Commentary was defending just the

opposite view. This is called "re-positioning" oneself. Good

sign...Carlos Sonnenschein, personal communication

Cancer, checkpoint inhibitors, and confusion.

Fernandes M, Brábek J.

Lancet Oncol. 2017 18(11):e632.

Compared to chemotherapy, serious broad spectrum toxicity is unpredictable.



medicines!



medicines!

Affordable cancer care.

Fernandes M, Brábek J.

Lancet Oncol. 2012 13(1):e2-3.

Affordable cancer care: pipedream or achievable reality?

Collingridge D, Sullivan R.

Lancet Oncol. 2014 15(3):257-8.

Continuing failure despite:

• excellent scientists

• advanced technology

• generous funding

WHY?

Not focused on central issue:

Invasion and metastasis!

Pharma companies don’t concentrate on solving the problem

of metastasis (the thing that kills people); they focus on

devising drugs that shrink tumors (the things that don’t).

In contrast to hematopoetic malignancies, solid

cancer is predominantly a disease of invasion

and metastasis, not an uncontrolled proliferation!

x

Virchows Archiv B

December 1978, Volume 29, Issue 1,

pp 145–150 | Cite as

Tumors as clonal proliferation

•Authors

•Authors and affiliations

•Peter C. Nowell

Cancer; a definition

A growth (enlargement) composed of a clonal population

of cells that has acquired the ability to expand in defiance

of the checks and balances that would normally control

the proliferation and survival of normal cells (neo-plastic).

https://link.springer.com/journal/443https://link.springer.com/journal/443/29/1/page/1https://link.springer.com/article/10.1007/BF02899348#citeashttps://link.springer.com/article/10.1007/BF02899348#authorshttps://link.springer.com/article/10.1007/BF02899348#authorsandaffiliations

In solid cancer, does ongoing metastatic activity

negate the “benefit” of tumor shrinkage?

YES!

METASTASES

➢ according to WHO - 9 millions of cancer deaths per year

(more than 10% of all deaths)

➢ metastases responsible for 90 % of deaths in cancer patients

(Steeg, 2016)

(adopted from (Hanahan a Weinberg, 2011))

HALLMARKS OF CANCER

Sustaining proliferative

signaling

Evading growth

suppressors

Enabling replicative

immortalityResisting cell death

Inducing angiogenesisActivating invasion

and metastasis

Avoiding immune

destruction

Deregulating cellular

energetics

Tumor-promoting

inflammation

Genome instability

and mutation

- Sustaining proliferative signaling

- Evading growth suppressors

- Resisting cell death

- Enabling replicative immortality

- Inducing angiogenesis

- Activating invasion and metastasis

MALIGNANT TRANSFORMATION

(Adapted from (Hanahan a Weinberg, 2000))

➢ invasion and metastasis - the only real

cancer hallmark

(prof. Y. Lazebnik)

➢ understanding the mechanisms of

invasion and metastasis critical for the

development of effective anti-cancer

treatment

➢ change in regulations required - criteria

of effectiveness for antimetastatic

drugs(Brábek and Fernandes, The Lancet Oncology, 2012)

METASTASES

© EU, 2016

(Pantel and Brakenhoff, 2004)

METASTATIC CASCADE

ECMBM

intravasation

invasionvessel

primary tumor

METASTATIC CASCADE

I. Invasion – from primary tumor into the

surrounding tissue.

II. Intravasation – into blood or lymphatic vessel.

III. Transport to the secondary site and survival in

the bloodstream.

IV. Extravasation at the secondary site.

V. Proliferation at the secondary site,

formation of metastases.

vessel

extravasation

vessel

dormancy metastasis

IndividualCollective

Mesenchymal

Amoeboid

MODES OF CANCER CELL INVASIVENESS

HT1080 cells

MESENCHYMAL INVASIVENESS

➢ elongated shape

➢ proteolytic degradation of ECM

➢ formation of invadopodia

➢ actomyosin stress fibers

➢ Rac signalization

➢ migration speed 0,1-1 µm/min

(Tolde, unpublished results)

DAPI actin p-paxillin

DAPI actin p-paxillin

Podosomes and invadopodia – ECM degradation(Mitra et al., 2005)

Focal adhesions – adhesions to ECM

(Block et al., 2008)

CELL STRUCTURES IMPORTANT FOR

ADHESION AND INVASIVENESS

(Tolde et al., 2010)

PODOSOME COMPONENTS

(Brunton et al., 2004)

INVADOPODIA – STRUCTURE

2D (glass):

F-actin

p-Tyr

3D – acellular dermis

Tolde 2010


INVADOPODIA


10 μm 10 μm

+ GM6001

+GM6001

DAPI

F-actin

FITC-dermis

0 -10 -20 -30 -40 µm

MESENCHYMAL INVASIVENESS IS DEPENDENT

ON PROTEOLYTIC ECM DEGRADATION


AMOEBOID INVASIVENESS

➢ round shape

➢ ECM proteases-independent

➢ elevated traction forces

➢ cortical actomyosin

➢ Rho/ROCK signalization

➢ migration speed 2-25 µm/min

(Gandalovičová, unpublished results)

HT1080-iRhoA (G14V)

AMOEBOID INVASIVENESS

Elevated Rho/ROCK signaling results in elevation

of traction forces in amoeboid cancer cells.

(Rosel et al., 2008)

K2 – mesenchymal A3 - amoeboidTraction

[Pa]

0

250

500

MLCP

ROCK

MRCK

MLCK

ZIPKp

p-MLCMLC

PAK1

actomyosin

contractility

Cdc42

RhoCRhoA

Rac

5 µm

AMOEBOID INVASIVENESS IS INDEPENDENT

ON ECM DEGRADATION

nuclei

F-actin

FITC-dermis

0 -10 -20 -30 -40 µm

+ GM6001

+GM6001

(Tolde, unpublished results)

+ ++ +++ +++

AMOEBOID INVASIVENESS - METASTASIS

THE IMPORTANCE OF RHO/ROCK/MLC

PATHWAY

AMOEBOID INVASIVENESS - METASTASIS

THE IMPORTANCE OF RHO/ROCK/MLC

PATHWAY

EMT : Epithelio-mesenchymal transition

MESENCHYMAL

EMT

Snail, Twist

E-Cadherin

COLLECTIVE

MAT : Mesenchymal-ameboid transition

AMT : Ameboid-mesenchymal transition

MAT

AMT

Proteases

Integrins

Rho/ROCK

AMOEBOID

➢ AMT/MAT represents an escape mechanism for cancer cell from treatment e.g. with protease inhibitors

CANCER CELL INVASION PLASTICITY

MESENCHYMAL TO AMOEBOID TRANSITION

(Gandalovičová, unpublished results)

HT1080-iRhoA (G14V)

Mesenchymal – amoeboid transition

Amoeboid – mesenchymal transitionA375m2 (melanoma) treated with p38 kinase inhibitors (10 μM)

DMSO SB202190 BIRB796

M2 macrophages in dense (4,8

mg/ml) or sparse (1,65 mg/ml)

collagen

Mesenchymal HT1080 cells after induction of RhoAG14V

or treatment with 1 μM dasatinib

4,8 mg/ml

1,65 mg/ml

(Cell invasion in cancer lab, unpublished results)

Transcriptomic and proteomic analysis of amoeboid mode

of invasiveness

HT1080-iRhoAG14V+ doxycyklin vs. HT1080 +dasatinib

545 117 762 717 219 759

TRANSCRIPTOMICS

Elevated expression in amoeboid cells Decreased expression in amoeboid cells

RhoAG14VRhoAG14V dasatinib dasatinib

Gene set enrichment analysis (GSEA)

– search for similarities with published

data

Interferones alfa/gama

activated genes (25/26)

TNFalfa activated genes (8)

G2/M checkpoint (41)

E2F target genes (40)

Mitotic spindle (25)

The adaptation of cell to amoeboid phenotype is associated

with the activation of inflammatory (JAK/STAT, NFkB)

signaling pathways and supression of cell cycle(Cell invasion in cancer lab, unpublished results)


of invasiveness


of invasiveness

Gene set enrichment analysis (GSEA)IL24

IL1B

IL11

CXCL8

IL1A

LIF

TGFA

GDF15

S100A16

INHBA

EPHB2

FST

VEGFA

GDF7

CLCF1

SPARC

EFNB2

BMP2

BMP8B

BMP7

GDNF

TGFB1

HGF

PDGFA

JAG1

Genes with decreased expression after AMT:

KEGG Gene Sets

„Cytokine-cytokine receptor interaction“ (25 genů)

„Jak-STAT signaling pathway“ (14 genů)

„MAPK signaling pathway“ (17 genů)

„TGF-beta signaling pathway“ (9 genů)

Hallmark Gene Sets

„genes regulated by NF-κB in response to TNF“ (50 genů)

„genes defining epithelial-mesenchymal transition“ (44

genů)

„genes up-regulated by KRAS activation“ (36 genů)

„genes up-regulated in response to hypoxia“ (33 genů)

„genes up-regulated by STAT5 in response to IL2

stimulation“ (32 genes)

„genes defining inflammatory response“ (27 genů)

„TGF-beta signaling pathway“ (9 genů)

„genes up-regulated by IL6 via STAT3“ (8 genů)

AMT in A375m2

results in decreased

expression of many

secreted ligands

HT1080

A375m2HT1080

Co-cultivation of mesenchymal

HT1080 cells with A375m2 cells in

mixed spheroids triggers amoeboid

invasiveness in HT1080 cells

Increased expression in mesenchymal cells: 2084 genes

Decreased expression in mesenchymal cells: 1769 genesA375m2 after p38 kinase

inhibition

(Cell invasion in cancer lab, unpublished results)

(Gandalovičová et al., 2017)

MIGRASTATICS – NEW CATEGORY

OF ANTI-CANCER DRUGS

MIGRASTATICS – NEW CATEGORY OF ANTI-

CANCER DRUGS

(Gandalovičová et al., 2017)

ANTIMETASTATIC TREATMENT– PROBLEMS

WITH CRITERIA OF EFFECTIVENESS

(Brábek et al., 2016)

FDA approves new treatment for a certain type of prostate

cancer using novel clinical trial endpoint

February 14, 2018

The U.S. FDA today approved Erleada (apalutamide) for the

treatment of patients with prostate cancer that has not spread,

but that continues to grow despite treatment with hormone

therapy. …This approval is the first to use the endpoint of

metastasis-free survival, measuring the length of time that

tumors did not spread to other parts of the body or that death

occurred after starting treatment,” said Richard Pazdur, M.D.,

director of the FDA’s Oncology Center of Excellence and acting

director of the Office of Hematology and Oncology Products in

the FDA’s Center for Drug Evaluation and Research.

"The FDA had now recognized that a prolonged delay in the

development of metastatic disease is an objective and

clinically relevant measure," Beaver and colleagues write.

THE CONCEPTUAL ADVANTAGES

OF MIGRASTATIC TREATMENT

cytotoxic challenge

Darwinian selection

of resistant clones

proliferative advantage

invasion/metastasis

no cytotoxic challenge

rare resistants without

proliferative advantage

NO invasion/metastasis

(Steeg, 2016)

INCORPORATION OF PRECLINICAL METASTASIS

MODELS INTO DRUG DEVELOPMENT

CONCLUSION:

PLAN B FOR CANCER MOONSHOT INITIATIVE

Plan A Precision medicine (”driver genes” etc.)

Plan B Therapy directed to ”3 Ms”

cancer microenvironment, motility (MIGRASTATICS) and metabolism

(Brábek, Rosel, Fernandes, New England Journal of Medicine, 2017)

WHAT´S NEXT

➢ Based on our work, we plan on exploring the determinants

of metastasis and tractable targets for pharma

intervention, e.g. migrastatics.

➢ Collaboration with medicinal chemists is crucial and we need

to explore both synthetic compounds and natural products.

➢ Accordingly, we plan on making suggestions for the revision

of regulations on new cancer drugs. Primarily, rationalizing

and justifying criteria for the evaluation of sustainable

efficacy.

“The basis for the allele-oriented SMT has recently been

questioned, and the relevance of embryonic fields to cancer

has been re-established. Alterations in paracrine factor

signaling in both the target and producer cells have been

seen to initiate cancer formation, and embryonic

processes such as epithelial-mesenchymal

transformation are now seen as critical in metastasis. It is

without question, though, that developmental biology helped

establish oncology and has continued to help mold it. The

rightful sovereign returns.”

(Joyce and Pollard, 2009)

TUMOR HISTOLOGY

STROMAL ENVIRONMENT INTERACTIONS

(Mueller a Fusenig, 2004)

(Vander Heiden, et al., 2009)

STROMAL ENVIRONMENT INTERACTIONS

(Olumi, et al., 1999)

Cancer-associated Fibroblasts (CAFs)

TUMOR MICROENVIRONMENT

CANCER CELL INVASIVENESS

THE ROLE OF CAFs

(Wolf et al., 2007)

Ctrl Ctrl - GFP Ctrl HFP3 HFP3+A2058 Mel Fib MelFib+A2058

+IL8 Antibody

CANCER CELL INVASIVENESS

THE ROLE OF CAFs

(Jobe et al., 2017)

PODĚKOVÁNÍ

DANIEL RÖSEL

ONDŘEJ TOLDE

VLADIMÍR ČERMÁK

ANETA GANDALOVIČOVÁ

LADISLAV MERTA

DANIELA PAŇKOVÁ

KAREL SMETANA Jr.

PAVEL MARTÁSEK

VLADIMÍR KRÁL

PAVEL VESELÝ

JAN KOSLA

MICHAL DVOŘÁK

JIŘÍ PLACHÝ

BEN FABRY

VICTORIA SANZ-MORENO

Biotechnology and Biomedicine Center

of the Academy of Sciences

and Charles University in Vestec

THANK YOU FOR ATTENTION!

Paradigm shift in solid cancer and the need for antimetastatic drugs · Affordable cancer care....

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