Giorgio Parmiani
description
Transcript of Giorgio Parmiani
Forlì, March 1-2, 2007
Anti-cancer vaccines
Peptide-based vaccines: The INT experience
Giorgio Parmiani, MD
Department of Oncology,
San Raffaele Scientific Institute
Strategies of anti-cancer therapeuticStrategies of anti-cancer therapeutic vaccinesvaccines
1. Irradiated and genetically modified tumor cells rradiated and genetically modified tumor cells
2. TAAs
3. Antigenic peptides with adjuvants or dendritic cells
4. Tumor-derived heat shock proteinseat shock proteins
- as whole proteins plus adjuvants
- as genes transduced in normal cells- as plasmid DNA- as DNA contained in viral or non-viral vectors- aS mRNA
Peptide-based vaccines
• Advantages:
• Known sequence and biochemistry
• Easy to synthesize (large availability)
• Allow a specific immune-monitoring of the patient response
• Allow to assess expression of targeted antigens in patient tumor cells
Peptide-based vaccines (cont.)
• Disadvantages:
• Easy degradable in absence of adjuvants
• Require appropriate HLA allele to be recognized (patient selection)
• Induce T cells that may not recognize tumor cells
• Costs
Issues in peptide-based cancer vaccines
• Nature of tumor-associated antigens• Vaccine formulation/adjuvants• Quantity and quality of immune response • First generation of vaccines (1994-
2004): Immmune and clinical response• Immune suppression/tumor escape• Regulatory• Costs
Tumor-associated antigens Tumor-associated antigens (TAA):(TAA):Their nature is crucial Their nature is crucial for anti-tumor immune for anti-tumor immune responseresponse
Antigens recognized by T cells Antigens recognized by T cells (Novellino (Novellino
et al., Cancer Immunol Immunother 2005):et al., Cancer Immunol Immunother 2005):
1)Shared, self differentiation proteins expressed also on normal cells(e.g. MART-1, CEA, PSA)
2)Shared self Cancer/Testis expressed by different tumors and by normal testis or placenta (e.g. MAGE, NY-ESO-1)
3)Shared but predominantly expressed on tumor cells (e.g. RAS/m, survivin, hTERT)
4) Unique, expressed only by a single tumor(e.g.CDK4/m,α-actin-m)
1 and 2.1 and 2. In vitroIn vitro andand exex--vivovivo T cellT cell
natural immune response to natural immune response to shared “self” TAAsshared “self” TAAs
0.0
0.1
0.2
0.3
0.30.81.31.82.32.83.3
n.s.***
0.0
0.1
0.2
0.3
0.30.81.31.82.32.83.3
n.s. *****
gp100209-217 Tyrosinase368-377 Melan-A/MART-126-35
Patients
Donors StageI-II
StageIII-IV
FREQUENCY OF SPONTANEOUS CIRCULATING ANTI-MAA CD8+FREQUENCY OF SPONTANEOUS CIRCULATING ANTI-MAA CD8+ T LYMPHOCYTES T LYMPHOCYTES ((Mortarini et al., Cancer Res 2004Mortarini et al., Cancer Res 2004))
0.0
0.1
0.2
0.3
0.30.81.31.82.32.83.3
Patients
Donors StageI-II
StageIII-IV
Patients
Donors StageI-II
StageIII-IV
*** ********
CONCLUSIONSCONCLUSIONS
• A hierarchy exists in the natural A hierarchy exists in the natural recognition ofrecognition of “self”“self” TAAs by T TAAs by T cells.cells.
• MelanA/MART-1 e CEA induce MelanA/MART-1 e CEA induce the most frequent T cell the most frequent T cell response in melanoma and CRC.response in melanoma and CRC.
• Recognition ofRecognition of “self” TAAs“self” TAAs
increases with the increased increases with the increased tumor burden.tumor burden.
Antigens recognized by T cells:Antigens recognized by T cells:1) Shared, self differentiation proteins expressed
also on normal cells (e.g. MART-1, CEA, PSA)
2) Shared self Cancer/Testis expressed by different tumors and by normal testis or placenta (e.g. MAGE, NY-ESO-1)
3) Shared but predominantly expressed on tumor cells (e.g. RAS/m, survivin, hTERT)
4) Unique, expressed only by a single tumor(e.g. CDK4/m, α-actin-m)
SURVIVIN (SVV)SURVIVIN (SVV)
• Expression pattern:• abundant during fetal development• silenced in normal adult tissues• over-expressed in most common
human cancers
• Member of AIPs (Apoptosis Inhibitory Proteins)
Survivin is a universal TAA that can elicit both HLA-I and HLA-II-restricted T cells
IN VITRO INDUCTION OF HLA CLASS I-RESTRICTED
ANTI-SVV T CELLS IN PBMCs OF RECTAL CANCER PATIENTS
0
25
50
75
100
- + - +
0
25
50
75
100
- + - +
Autologous PBMCs
SVV-1Medium
IFN
γ s
pots
/50,0
00 c
ells Patient 1
Patient 2
+ anti-HLA-I Ab
(Casati et al., Cancer Research 2003)
0
25
50
75
100
med
PSMA1
PSMA2
svv
Her2neu
EpCam
Flu
N.
IFN
gam
ma
spo
ts/2
x10
5 P
BM
Cs
In vitro recognition of prostate antigens-deriving peptides byT lymphocytes of prostate cancer patients
In vitro recognition of prostate antigens-deriving peptides byT lymphocytes of prostate cancer patients
Antigens recognized by T cells:Antigens recognized by T cells:1) Shared but expressed on tumor cells
only
2) Shared Cancer/Testis expressed by different tumors and by normal testis or placenta
3) Shared, differentiation proteins expressed also on normal cells
4)Unique, mutation-derived,expressed only by a single tumor (e.g. CDK4/m, α-actin-m)
Unique human TAAs recognized by HLA-restricted T cells (Parmiani et al.,
Journal of Immunology 2007)
Tumor No. Class I No. Class II
Melanoma 11 6NSCLC 3 1H/N 2 0RCC 2 1Bladder cancer 1Total 19 8
Most of these unique TAA have a pathogenetic role in neoplastic transformation/ progression.
Strategies of anti-cancer therapeuticStrategies of anti-cancer therapeutic vaccinesvaccines
1. Irradiated and genetically modified tumor cells rradiated and genetically modified tumor cells
2. TAAs
3. Antigenic peptides with adjuvants or dendritic cells
4. Tumor-derived heat shock proteinseat shock proteins
- As whole proteins plus adjuvants
- As genes transduced in normal cells- As plasmid DNA- As DNA contained in viral or non-viral vectors- AS mRNA
Use of shared “self” or “cancer/testis” TAA peptides as
vaccines in patients with metastatic melanoma
(1995-2004). Clinical RR 18%.
CD8+T cell response in20-50% of patients but
limited expansion of T cells (<1%).
Phase I/II studies of DC-peptide based Phase I/II studies of DC-peptide based therapeutic vaccines in melanomatherapeutic vaccines in melanoma
DC+ antigen No.Pts Responses Ref. Immunologic Clinical
Peptides1 or lysate 32 11 2 CR, 6 PR Nestle 2001
HLA-A2 peptides1 18 16 3 CR Banchereau 2001
Tumor lysate 11 5 1 PR Mulé 2002
Tumor lysate 17 5 2 CR, 3PR Ridolfi, 2006
MAGE-DR 16 15 1 CR Schuler 2002
Gp100, tyrosinase 12 1/9 1 PR Slingluff 2003
Melan-A + IL-12 20 5 2 CR Gajewski 2003
Gp100-9V 12 12 2 PR Haluska 2005
Allo-Tumor cells 13 3 1CR, 1PR Banchereau 2006
Total 151 73 (51%) 25 (13%)
1 Peptides included MART-1, MAGE, tyrosinase, gp100.
Rationale for a new generation of peptide-based vaccines at INT of
Milan
• Multiple peptide including universal TAAs
• New TLR targeting adjuvants (CpG, HSP)
• Down-regulation of Tregs and/or Myeloid Suppressor Cells (see Licia Rivoltini)
• Immune-monitoring in blood and LNs
• Assessment of patient polymorphisms
On the basis of pre-clinical results, the SVV-1.HLA-A2 peptide is being used, along with other peptides, in clinical studies of vaccination of patients with prostate and rectal carcinoma and melanoma.
Vaccination
HLA-A2-restricted peptides from PMSA1, PSMA2, survivin + Montanide. CPA before vaccination
Vaccination
HLA-A2-restricted peptides from PMSA1, PSMA2, survivin + Montanide. CPA before vaccination
Vaccination with peptides deriving from prostate TAAs in association with Montanide and cyclophosphamide (CPA)
Peptides
Activation ofspecific T cells
Unit of Immunotherapy of Human Tumors
CPA(300 mg/m2)
Blocking of suppressive effectof CD4+ CD25+T regon lymphocyte proliferation
Montanide
Recruitment of pro-infliammatory cells(APC)
Vaccination with peptide deriving from prostate TAAs in association with Montanide and CPA
Group 1 – Adjuvant Vaccination
•Malattia localizzata a livello prostatico, resecabile•Vaccinazione in regime neoadiuvante ed adiuvante dopo chirurgia
Unita’ di Immunoterapia dei Tumori Umani
1° cycle vaccine 2° cycle vaccine
C C-3 1 w
C C-3 1 month
C C
Diagnostic biopsy
ProstatectomyProstatectomy
Analysis of induction of T lymphocytes recognizing vaccine peptides in PBLs
Analysis of tumor infiltrate
Results: Immunologic and clinical responses
Vaccination of prostate Vaccination of prostate cancer patients: Clinical cancer patients: Clinical
responseresponsePatientPatientss
Clinical outcomeClinical outcome
001 Biochemical SD at 10 months from interruption of hormone therapy
002 Biochemical SD at 8 months from interruption of hormone therapy
003 Biochemical SD at 6 months from interruption of hormone therapy
004 Biochemical SD at 6 months and CR of a prostatic lesion
005 PD
006 PD
007 After the first cycle of vaccination (PSA 1.33 ng/ml → 1.01 ng/ml)
008 Aftrer the first cycle of vaccination (PSA 0.51 ng/ml → 0.44 ng/ml)
009 After the first cycle of vaccination (PSA 1.04 ng/ml → 0.27 ng/ml)
Vaccination of high risk stage IIB/IIIC melanoma patients with multipeptides
emulsified in Montanide , and given cyclophosphamide and IL-2 vs.
observation only. A randomized phase II study
• G. Parmiani, M. Santinami, L. Pilla, G. Cutolo, A. Maurichi, R. Patuzzo, A. Carbone, G. Tragni, C. Castelli, L. Rivoltini (INT);
• M. Maio (Siena),V. Chiarion (Padua)
• Ethics Committee approval: May 2004.
• Delayed due to the long term negotiation with Coley to obtain CpG-ODN (not provided).
• First patient enrolled in July 2005
Primary end points: Immune response (frequency and intensity)
ELISPOT: Frequency of anti-tumor specific T lymphocytes
Flow-cytometry: Phenotype of anti-tumor T cells
-HLA tetramers-Cytokine intracellular staining
-Immunohistochemistry: characterization of tumor cells and of TILs
Clinical objectives
Primary:
Local and systemic toxicity
Secondary:
Disease-free survival after 2/3 years
Overall survival at 5 years
HLA- A2
negative
patients
60%
HLA- A2
positive
patients
40%
No. of assessed patients: 210
ENROLLED PATIENTS: 28
Stage II: 6 Stage III: 22
Control arm: 13 Vaccination arm: 15
SVV as universal TAA : - Conclusions -
• SVV is expressed by melanoma; colon, rectal and prostate cancer cells
• Class I and II HLA-restricted SVV epitopes can be recognized both ex-vivo and in vitro by patient T though to a variable extent
• SVV can then be considered a shared, universal tumor-restricted antigen that is being used in clinical studies of vaccination
Antigens recognized by T cells:Antigens recognized by T cells:1) Shared but expressed on tumor cells
only
2) Shared Cancer/Testis expressed by different tumors and by normal testis or placenta
3) Shared, differentiation proteins expressed also on normal cells
4)Unique, mutation-derived,expressed only by a single tumor (e.g. CDK4/m, α-actin-m)
Vaccination withVaccination withunique antigens:unique antigens:
how is it possible?how is it possible?1.Autologous gene-modified tumor 1.Autologous gene-modified tumor
cellscells
2. Autologous 2. Autologous proteins/peptidesproteins/peptides
Autologous tumor-derived
heat shock-protein/peptides complexes
MECHANISMS OFMECHANISMS OFIMMUNE EVASIONIMMUNE EVASION
Why a more strong Why a more strong immune immune responseresponse and/or a more frequent and/or a more frequent
clinical responseclinical response was not observed was not observed in vaccination trialsin vaccination trials
??
FACTORS THAT CAN INTERFERE WITH THE RECOGNITIONFACTORS THAT CAN INTERFERE WITH THE RECOGNITIONAND LYSIS OF TUMOR CELLS BY T LYMPHOCYTESAND LYSIS OF TUMOR CELLS BY T LYMPHOCYTES
TumorTumor Lack of antigenic peptide or
down-regulation of HLA
Dysfunction of antigen presentation
Release of immune-suppressive factors (IL-10, TGF, VEGF)
Tumor counterattack (Fas/FasL)
Cannibalization of T lymphocytes
Immune systemImmune system Immune anergy or
ignorance
Lack of tissue homing molecules; defective adhesion
TCR dysfunction
Inactivation of T cells within the tumor (granzyme B)
Induction of anti-peptide but not anti-tumor T cells
T regulatory cells
Myeloid suppressive cells
AcknowledgmentsAcknowledgmentsChiara CastelliLicia RivoltiniChiara Camisaschi Gianluca CutoloVeronica Huber Manuela Iero
Andrea MarrariLuisa NovellinoLorenzo PillaRoberta Valenti
Agata CovaFrancesca Rini Gloria SovenaPaola Squarcina Istituto Nazionale Tumori
Milan - Italy
Vincenzo MazzaferroJorgelina CoppaMario SantinamiRoberto PatuzzoErmanno LeoFiliberto BelliFlavio ArientiAlfonso Marchianò
Grazia BarpAnnabella Di Florio
Pramod K. SrivastavaAntigenics, New York
Michele MaioSiena
Take home message. Perspectives 1
• Antigens: use both cross-reacting self and unique TAA; use both classes I and II HLA-restricted peptides.
• Adjuvants: CpG-ODN plus IFA are likely to be the most potent; cytokines: how to use them?(IFN-, GM-CSF?, IL-12, IL-15, IL-21, IL-23).
• Tumor burden: avoid excess tumor burden to prevent release of suppressive factors.
1
Perspectives 2
• Escape mechanisms: use compounds which may help in restoring normal immune functions(e.g. retinoic acid, cytokines, anti-Treg Ab).
• Combination therapy: vaccination may be combined with anti-angiogenic drugs, radiotherapy and immune-modulating chemotherapy(non-myeloablative lymphodepletion?)
• Modulation of the immune response by anti-Treg and/or anti-CTLA4 antibodies.
2
REGULATORY T LYMPHOCYTES
Treg assessment during peptide-based vaccination and
modulation by Cyclophosphamide
• Recurrent rectal carcinoma patients receiving a peptide-based vaccine(no IL-2)
• Stage III melanoma patients receiving a multipeptide-based vaccine (with IL-2)
4
6
8
10
12
14
CTX1 V1 CTX2 V4 CTX3 V5 post
8
9
10
11
12
13
CTX1 V1 CTX2 V4 CTX3 V5 post
7
8
9
10
CTX1 V1 CTX2 V4 CTX3 V5 post
8
9
10
11
12
13
14
CTX1 V1 CTX2 V4 CTX3 V5 post
% o
f C
D25
+ F
oxp
3+ (
in C
D4)
Pt#4 Pt#3
Pt#2Pt#1
Treg (CD4+CD25+Foxp3+) in PBMC of rectal carcinoma patients and their modulation by CTX treatment
10
15
20
25
30
35
CTX1V1
Surge
ryCTX2
V3 V4
post
IL-2
5
10
15
20
CTX1V1
Surge
ryCTX2
V3 V4
post
IL-2
5
10
15
20
CTX1V1
Surge
ryCTX2
V3 V4
post
IL-2
% o
f p
os
itiv
e ce
lls
CD4+CD25+ CD25+Foxp3+(in CD4) CD4+CD25brigh
Pt#1 Pt#2
Treg in PBMC of Melanoma patients: modulation by CTX and IL-2 treatment
Studi clinici di vaccinoterapia in assenza di cellule dendritiche
in pazienti con carcinoma prostaticoVaccino N. HLA Risposta Risposta Ref.
pazienti immune clinica
SART, p56LCK, 10 A2402 4/10 1 PR* Naguchi, ’03ART, CyB 5 SD
SART, p56LCK, 16 A2402 10/14 13/13 ** Naguchi, ’05ART, CyB, PSA, PAP, (71%) (6/13 PR)PSMA estramustine
Allo-prostata + BCG 26 A*0201 11/26 11/26§ Pandha, ’05
pVAX/PSA + GM-CSF 9 A*0201 2/9 Non Pisa, ‘04+ IL-2 valutabile
Auto-Prostate/GM-CSF 8 NV 7/8 Non Simons, ’99 DTH valutabile
FowlPox+VacPSA 64 NA 46% 18 PFS Kaufman ‘04 (78%)
* 89% PSA - ** Tutti PSA, con 6/13 > 50%.§ Riduzione prolungata e statisticamente significativa di PSA.
Features of peptide vaccines
• Peptide length:
• 8-10 aa for HLA class I-restricted peptides
• 12-23 aa for HLA class II-restricted peptides
• Modified peptides (heteroclitic and/or long peptides)
• Highly and rapidly degradable in vivo (adjuvants are required)
1, 2: Shared “self” TAAs• Normal subjects and cancer patients show
some forme of tolerance to “self” TAAs (immune ignorance, peripheral tolerance, central tolerance, low frequency of T cells, etc.).
• Tolerance needs to be broken in order to induce a T cell immune response against “self” TAAs.
• Thus, these TAAs are considered to be “weak antigens”
Natural CD8+T cell response of HLA-A2 colorectalNatural CD8+T cell response of HLA-A2 colorectalcancer (CRC) patients to differentiation antigenscancer (CRC) patients to differentiation antigens
(Nagorsen et al., Cancer Res 2000)(Nagorsen et al., Cancer Res 2000)
Antigen Clinical N. of respondingstage N. of tested*
Ep-CAM Donors 0/8A-B 0/9C-D 4/13
Her2/neu Donors 0/8A-B 0/9C-D 5/13
CEA Donors 0/8A-B 0/9C-D 6/13
* ELISPOT assay.
0
25
50
75
100
N.
IFN
γ s
pots
/3x1
04
cells
med
T2+
PS
MA
T2+
PS
MA
+ a
nti
-HL
A-I
CaP
+ a
nti
-HL
A-I
CaP
Anti-PSMA T cells
CaP
Anti-svv T cells
0
25
50
75
100
med
T2+
svv
T2+
svv
+ a
nti
-HL
A-I
CaP
+ a
nti
-HL
A-IN.
IFN
γ s
pots
/3x1
04
cells
Fresh PBMC
0
25
50
75
100
med
PS
MA
sv
v
Flu
N.
IFN
γ s
pots
/10
6 c
ells
A
B
A. Analisi della frequenza di precursori linfocitari T anti-tumore specifici nel sangue periferico in pazienti con carcinoma della prostata. I PBMC isolati attraverso centrifugazione in gradiente di densità dal sangue periferico di pazienti affetti da carcinoma della prostata HLA-A*0201 sono stati incubati con peptidi di PSMA, survivina e dalla proteina del virus influenzale (M1), come controllo positivo. Il riconoscimento dei vari antigeni da parte dei linfociti e’ stato valutato come rilascio di IFNγ, mediante Elispot. I dati rappresentano 1 dei 2 casi di pazienti con carcinoma prostatico che hanno mostrato un’elevata immunogenicita’ per gli antigeni analizzati.
B. Analisi quantitativa dell’attivazione di linfociti T anti-PSMA-specifici derivati da pazienti affetti da carcinoma prostatico. I PBMC di pazienti HLA-A*0201 sono stati stimolati per due settimane con i peptidi tumorali derivanti da PSMA e survivina per due settimane. Al termine di tale periodo la specificità dei linfociti T nei confronti di cellule T2 e di cellule di carcinoma prostatico LNCaP (?), è stata valutata quantificando il rilascio di IFNattraverso analisi immunoenzimatica Elispot.
1 2 3 54 6 1 2 3 54 67 8 9 10 7 8 9 10
T2 + Survivin T2 + Survivin + anti-HLA-I
T2 + PSMA P1T2 + PSMA P1 + anti-HLA-I
N.
of
sp
ots
per
30
4 c
ells
N.
of
sp
ots
per
30
4 c
ells
Frequency of TAA-specific T cells precursorsafter in vitro stimulation with prostatic peptides
No. of microcultures tested/patient
Ex-vivo recognition of prostate antigens by patient T cells
• No. of patients tested: 112
• No. of patients whose peripheral blood T lymphocytes recognized prostate antigens: 65 (58%)
Studio di fase III di vaccinazione con DC di Studio di fase III di vaccinazione con DC di pazienti con carcinoma della prostatapazienti con carcinoma della prostata
N. Pazienti Antigene Risposta Risposta P.I.immune clinica
110 vs 108 Provenge* ND Mediana E.Small, Metastatici, vs placebo sopravvivenza 2005ormone 25.9 vs 21.9resistenti OS 34% vs 11%
ND: non disponibile*Provenge= Proteina di fusione PAP+GM-CSF su cellule dendritiche
Patient 8- 3R
MHC-CLASS II RESTRICTED RESPONSE OF COLORECTAL MHC-CLASS II RESTRICTED RESPONSE OF COLORECTAL
CANCER PATIENTS TO rSVV PROTEIN OR CANCER LYSATECANCER PATIENTS TO rSVV PROTEIN OR CANCER LYSATE
IFN- ELISA
0
500
1000
1500
2000
2500
IFN
- p
g/m
l
Patient 1- 3R
IFN
- p
g/m
l
= medium
= auto LCL
= HCT 116 lysate
= melanoma lysate
= LCL + rSVV
= LCL + rSVV + L243
= LCL + HCT 116 lysate
= LCL + HCT 116 lysate + L243
= LCL + C1R/SVV lysate
= LCL + C1R/SVV lysate + L243
0
500
1000
1500
2000
2500
(Casati C et al., Cancer Research, 2003)
The response of autologous T cells to a human melanoma is dominated by
mutated neoantigens
(Lennerz V et al., PNAS, 2005)
• Five neoantigens deriving from somatic mutations were found at different time points in different metastases of a melanoma patient.
• CD8+ cytotoxic T cells against mutated TAA were predominant over shared differentiation self TAA over 4-5 years of testing.
Dendritic cells (DC)-based peptide vaccines
Characteristics:
* Direct stimulation of T cells in lymph nodes.
* Ex-vivo preparation of high numbers of DCs
and their phenotipic and functional
characterization.
* High cost and time consuming.
Are unique TAAs spontaneously recognized in vivo ?
T lymphocytes recognizing unique TAAs have been described in long-term surviving patients previously
affected bymelanoma; NSCLC;
head/neck, bladder and renal carcinoma
(T. Wolfel, P. Coulie, G. Parmiani, F. Triebel)
Phase I-II studies ofDC based therapeutic vaccines in prostate
cancerDC loaded with No. % Immune Clinical % Reference
pts response response
PAP/GM-CSF 31 38 3 (↓PSA) 10 Small ’00
PSMA/A2 37 NE 1 CR, 10 PR 31 Murphy ’99
PSA/mRNA 13 100 3 CR 23 Vieweg ’02
Mouse PAP 21 49 6 SD 0 Fong ’01
PSA 24 45 11 (↓PSA) 46 Barrou ’04
hTERT/mRNA 20 95 9 (↓PSA) 45 Vieweg ‘05
T O T A L 142 46 23
Autologous monocytes pulsed with Autologous monocytes pulsed with melanoma-derivedmelanoma-derived HSP70HSP70 are recognizedare recognized
by specific T cell clones by specific T cell clonesIF
N-
(p
g/m
l)
0
1000
2000
3000
Anti-gp100
APC
Med
ium
HS
P7
0
HS
P7
0+
W6
.32
Au
to-M
e
0
250
500
Anti-MART-1
APC
Med
ium
HS
P7
0
Au
to-M
e
Statistically significant inhibition.
HS
P7
0+
W6
.32
W6.32 = anti-HLA-I mAb Auto-Me = autologous melanoma
(from Castelli et al., Cancer Res. 2001)
Some recent good news in
anti-cancer vaccination
Recombinant NY-ESO-1 protein with ISCOMATRIX
adjuvant induces broad integrated antibody and CD4+ and CD8+ T cell responses in melanoma
Davis et al., (Old LJ, Cebon JS), PNAS 101: 10697-10702, 2004
Treatment Disease-free survival at median follow-up of 2
years*
Placebo 2/7(28%)NY-ESO-1 protein only 7/16 (48%)NY-ESO-1 + IMX 17/19 (89%)
- Clinical results -- Clinical results -
* Well balanced for pathological state, primary lesion thickness, etc.
Phase II study of vaccination of hormone-refractory metastatic prostate cancer patients
with GVAX®Cell Genesys, ASCO 2005
• No. of patients 22• Median survival time of vaccinated patients was
over 29 months.• Median survival time of Taxotere-treated patients
is 19 months.• Phase III trials ongoing.
A phase IIb randomized study of vaccination with L-BLP25 in stage IIIb and IV NSCLC
after first line chemo
• Vaccine: Liposome-MUC1
• Overall median survival: 17.4 vs. 13 months in
88 vs. 83 patients
• Overall 2 years survival: 43.2 vs. 28.9 %;
subset stage IIIb: 60 vs.37 %
Biomira/Merck KGaA, ESMO 2004
Phase II randomized trial of autologous
formalin-fixed tumor vaccine for postsurgical recurrence of hepatocellular carcinoma
Kuang et al. Clin Cancer Res 2004
• No. of patients 41, after curative resection• Vaccine treatment N=19; no treatment N=22• Median follow-up of 15 months• Recurrence: Vaccinated 3/18 vs. Control 13/21• Recurrence-free survival P=0.003; OS P= 0.01
Istituto Nazionale Tumori Milano
Vaccinazione con 4 Peptidi emulsionati in
Montanide (MART-1, gp100, NY-ESO, survivin) allo scopo di diminuire l’emergere di varianti antigene negative
Razionale
Cancer Vaccine Pipeline(www.bioseeker.com)
• There are more than 260 non-antibody based vaccine drugs, therapeutic and preventive vaccines included. 87 are cancer vaccines.
• Vaccines are being developed for over 30 different cancers; 38 companies are involved.
• The number of vaccine targets has increased to over 50 different molecular targets
• Vaccine market is predicted to be of 6 billions USD by 2010
Clinical studies of vaccinationClinical studies of vaccination with HSPPC-96 with HSPPC-96
1.1. Liver metastases of colon Liver metastases of colon cancercancer
2.2. Metastatic melanomaMetastatic melanoma(HSPPC-96 only)(HSPPC-96 only)
3.3. Metastatic melanomaMetastatic melanoma(HSPPC-96 + GM-CSF + IFN)(HSPPC-96 + GM-CSF + IFN)
Vaccination of patients with liver metastases of CRC Vaccination of patients with liver metastases of CRC with autologous HSPPC-96with autologous HSPPC-96
- - TREATMENT SCHEMETREATMENT SCHEME - -
In vivoIn vivo expansion of anti-CEA T cells in HLA- expansion of anti-CEA T cells in HLA-A2 CRC patients vaccinated with HSPPC-96A2 CRC patients vaccinated with HSPPC-96
CD8
2.14
HL
A/ C
EA
tet
ram
ers
0.22
CD8
HL
A/ C
EA
tet
ram
ers
Day 1------7-------14------21 V1 V5
Patient 045Patient 045
N. s
pots
/104 T
cel
ls
V1 V5
*0
100
200T2 alone
T2+CEA
T2+CEA+W6.32
Colo206
0
100
200
300
400
500T2 alone
T2+CEA
T2+CEA+W6.32
Colo206
N. s
pots
/5x1
04 T c
ells
CD8CD8
Day 1---7—-14—21-------51 V1 V6
HL
A/ C
EA
tet
ram
ers
0.11 2.31
V1 V6
*
HL
A/ C
EA
tet
ram
ers
Patient 038Patient 038
Resection Purification of
HSP-GP96
Vaccination
Vaccination of metastatic patients with HSPPC-96 Vaccination of metastatic patients with HSPPC-96 derived from the autologous tumorderived from the autologous tumor
Tumor sample
Tumor cell suspension
Immunologic monitoringin vivo: DTHin vitro: ELISPOT, tetramers staining
VACCINATION WITH HSPPC-96 IN VACCINATION WITH HSPPC-96 IN METASTATIC COLORECTAL METASTATIC COLORECTAL
CARCINOMACARCINOMA(CRC)(CRC)
Clinical and immunologic responsesClinical and immunologic responses
(Mazzaferro et al., Clin Cancer Res 2003)(Mazzaferro et al., Clin Cancer Res 2003)
DISEASE-FREE SURVIVAL ACCORDING TO T CELL IMMUNERESPONSE IN PATIENTS WITH LIVER METASTASES FROM CRC
17 14 11 7 616 12 9 7
12 1 19 1 1
p < 0.0001
Immune responders
Immune non-responders
Patients at risk
Immune respondersImmune non-responders September 2004
OVERALL SURVIVAL ACCORDING TO T CELL IMMUNE RESPONSE
IN PATIENTS WITH LIVER METASTASES FROM CRC
17 17 17 14 11
Patients at risk
17 17 17 14Immune responders 12 9 6 311 6 5 2
p < 0.0001
2
Immune responders
Immune non-responders
Immune non-responders September 2004
HSPPC-96 vaccination increases CD8+T cell HSPPC-96 vaccination increases CD8+T cell response to class I HLA-matchedresponse to class I HLA-matchedCRC lines in 16 out of 29 patientsCRC lines in 16 out of 29 patients
0
50
100
150
200
250
300
350
400
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29
Pre Post (p < 0.05) Post (p > 0.05)501 537 698 449
N.
spots
/1.6
7 x
10
5 P
BM
C
Patient number
CONCLUSIONSCONCLUSIONSVaccination of patients with liver metastases of CRC with autologousHSPPC-96 is feasible and safe.
Approximately 60% of patientsdeveloped a T cell response to CRC
antigens and CRC cells.
Both OS and DFS were longer in T cell responding than in non-responding subjects,independently from other
prognostic factors.
TUMORS FOUND TO BE INFILTRATED BYT REGULATORY LYMPHOCYTES
Tumor Reference
NSCLC Woo/June, 2001
Pancreatic/breast cancer Liyanage, 2002
Colorectal cancer D. Herlyn, 2002
Gastric/esophageal cancer Ichihara, 2003
Melanoma Javia/Rosenberg, 2003
Ovarian Cancer Curiel, 2004
HCC Ormandy, 2005
Breast cancer Bates, 2006
Giorgio Parmiani
Istituto Nazionale Tumori, Milan - Italy
Mario SantinamiLorenzo Pilla
Roberto PatuzzoElisabetta Pennacchioli
Alfonso Marchianò
Licia Rivoltini Chiara Castelli
Phase III studiesPhase III studies of DC-based vaccines of DC-based vaccinesin patients with melanoma or prostate carcinoma in patients with melanoma or prostate carcinoma
No. Patients Antigen Immune Clinical P.I.response response
55 vs 53 MAGE1,3; Not available No advantage Schadendorf, 2006* Stage IV MART1, gp100,
tyrosine+DTIC vs DTIC
110 vs 108 Provenge** Not done Median Small, 2005Metastatic, vs placebo survivalhormone 25.9 vs 21.9resistant OS 34% vs 11%
**Provenge (Dendreon) = Fusion protein of PAP+GM-CSF loaded on autologous DCs.
*But in the vaccinated arm, a significant benefit in OS was found in the subset of HLA-A2+/B44 patients vs.non-HLA-A2/B44 ones.
DC-BASED VACCINATIONIN PATIENTS WITH METASTASES
- C O N C L U S I O N S -
• DC-peptide based vaccines induced a frequency of CD8+T cells response higher (up to 100%) than that obtained by peptide only-based vaccines but without a parallele increase in the clinical outcome (tumor response).
Istituto Nazionale Tumori Milano
Vaccination with 4 peptides emulsified in Montanide (MART-1, gp100, NY-ESO, survivin) with the aim of reducing the emergence of antigenic negative variants.
Rationale
Cyclophosfamide: 300 mg/m2: with the aim of down-regulating T regs
IL-2: 3x106 IU (s.c.): to expand T cells activated by the vaccine
67%9%
24% Others
Stage IIB/C
Stage IIIB
PATIENTS ENROLLED