Therapeutic Vaccines in

Cancer

Kunle Odunsi, M.D., Ph.D.1,2

Department of Gynecologic Oncology1

Center for Immunotherapy2

Roswell Park Cancer Institute

Buffalo, NY

Model of immune recognition of human

cancers

Circulation

Tumor

Lymph node

Traffic

CD8+ T cell

CD8+

T cell

APCTumor cell

Antigen

Tumor

Blood

vessel Memory T cell

• Summarize results from on-going vaccine studies utilizing NY-ESO-1 as target antigen in human ovarian cancer.

• What are the dominant mechanisms of immune escape in human cancers?

- Programmed Death-1 (PD-1) and Lymphocyte Activation Gene-3 (LAG-3), that result in T cell “unresponsiveness” in human ovarian cancer.

Objectives

0 20 40 60 80 100 120 140

Overall Survival (Months)

0.0

0.2

0.4

0.6

0.8

1.0

Cu

mu

lati

ve S

urv

ival

Intraepithelial CD8+ TIL

lowest tertile

all others

Log Rank test P=0.009

Median survival: 55 Vs 26 months

Hazard ratio: 0.33 (p = 0.0003)

Sato et al, PNAS. 2005, 102:18538

What are the targets of

immune recognition in human cancers?

• Categories of TA:

Differentiation antigens:

tyrosinase, MART1/melan-A, NY-BR-1.

Mutational antigens : ras, p53.

Over-amplified antigens: HER-2/neu, p53, WT-1, CD20.

Viral antigens such as HPV-16, 18-derived E6 and E7.

Cancer/Testis (CT) antigens:NY-ESO-1, MAGE, SSX, OY-TES-1, AKAP3, XAGE.

Potential Contribution of Antigen Specific

CD8+ T cells: NY-ESO-1

• Focal, 1+ to 4+ immunostaining

• Expression frequency 43% (n = 190)

• Humoral response: 30%

• Discovered by serological screening of a recombinant cDNA.

expression library obtained from an esophageal tumor (SEREX)

• Expression limited to germ cells and tumor cells

• Immunogenic

0 20 40 60 80 100 120 140

Overall Survival (Months)

0.0

0.2

0.4

0.6

0.8

1.0

Cu

mu

lati

ve

Su

rviv

al

NY-ESO-1 CD8 TIL

NY-ESO-1 (-) CD8+TIL(-)

NY-ESO-1(-)CD8+TIL(+)

NY-ESO-1(+) CD8+TIL(-)

NY-ESO-1(+) CD8+TIL(+)

Odunsi et al. Cancer Res. 2003, 63:6076)

Sato et al, PNAS. 2005, 102:18538

Efficacy of vaccination with recombinant vaccinia and fowlpox vectors expressing

NY-ESO-1 antigen in ovarian cancer and melanoma patients. Odunsi et al, PNAS.

2012, 109:5797

Vaccination with an NY-ESO-1 peptide of HLA class I/II specificities induces

integrated humoral and T cell responses in ovarian cancer.

Odunsi et al, PNAS. 2007, 104:12837

RPCI Ovarian Cancer Vaccine Program

I125207: Phase I Study of ALVAC(2)-NY-ESO-1(M)/TRICOM (vCP2292) in

Patients with Epithelial Ovarian, Fallopian Tube or Primary Peritoneal

Carcinoma Whose Tumors Express NY-ESO-1 or LAGE-1 Antigen (n=12)

(Unpublished).

I13303: A Phase II study of rV-NY-ESO-1 and rF-

NY-ESO-1 in patients with ovarian cancer whose

tumors express NY-ESO-1 or LAGE-1 antigen

Day 01 29 57 85 113 141 169 197

(n=22)

rVNY-ESO-1

3.1x107 PFUid

rFNY-ESO-1

7.4x107 PFU sc

Hypothesis: A diversified prime and boost vaccine regimen using two

different pox-viral vaccines would lead to enhanced CD4 and CD8 T cell

activation and sustained response against ovarian tumor.

Objective(s): To establish that the time to failure (ttf) for the proposed therapy is greater than the ttf for standard therapy.

Summary of Protocol I13303: Diversified prime boost rV-

NY-ESO-1 and rF-NY-ESO1 in ovarian cancer patients

(n=22)

Antibody responses:

• 3/22 (14%) baseline seropositives

• 8/19 (42%) seroconverted.

• Total ab responses: 11/22 (50%)

CD8 T cell responses:

• 3/22 (18%) with pre-existing CD8+ T cells

• 7/19 (42%) developed de novo CD8+ T cell

responses.

CD4 T cell responses :

• 9/22(40%) with pre-existing CD4+ T cells

• 7 additional patients developed de novo CD4+ T cell

responses.

LogRank Pval= <.001

Immune Category 1:No AB, CD4 or CD82:No AB. >=1 CD4 or CD8 Post

3:No Pre AB. Post AB.4:Pre and Post AB

Dis

trib

utio

n F

un

ctio

n

0.00

0.25

0.50

0.75

1.00

Time at Risk (Months)

0 12 24 36 48 60 72

Overall Survival by Immune Response Category

Median 53 mths

Median 14 months

Median 48 months

GOG 182: OS 40mths

The expression of each of the structural genes is driven by a

vaccinia virus promoter

Amino acid at position 165 of NY-ESO-1 changed from cysteine

to valine.

GM-CSF days 1-4 of vaccination.

ALVAC(2)-NY-ESO-1(M)/TRICOM

NYESO-1 NYESO-1H6H6

C6 C3C3

C5 C5

hICAhLFA- hB7. hICA hLFA-hB7.30K 30K

I3 I3H6sE/L sE/LE3L

ALVAC(2)-NYESO-1-TRICOM (vCP2292)

K3 E3

Protocol I125207: Phase I study of ALVAC(2)-NY-ESO-1(M)/TRICOM

in patients with ovarian cancer whose tumors express NY-ESO-1 or

LAGE-1 antigen

Month 1 Month 2 Month 3 Month 4 Month 5 Month 6

Legend:

ALVAC(2)-NY-ESO-1(M)TRICOM Vaccine SC once monthly

GM-CSF 100mcg/day Day 0-4 SC

1

10

100

1000

10000

100000

1000000

pre d29 d57 d85 d113 d141 d169

Days

Tit

er

#001

#002

#003

#004

#005

#006

#007

#008

#009

#010

#011

#012

Antibody responses for NY-ESO-1

p1

-20

p11

-30

p21

-40

p3

1-5

0

p41

-60

p51

-70

p61

-80

p71

-90

p81

-10

0

p91

-11

0p

10

1-1

20

p111

-130

p11

9-1

43

p13

1-1

50

p13

9-1

60

p15

1-1

70

p16

1-1

80

1

2

3

4

5

6

7

8

9

10

Pat. No.

CD8

po

ol

11

12

N/A

N/A

N/A

CD4

p1

-20

p11

-30

p2

1-4

0

p3

1-5

0

p4

1-6

0

p5

1-7

0

p6

1-8

0

p7

1-9

0

p8

1-1

00

p91

-11

0

p1

01

-12

0

p111

-130

p11

9-1

43

p1

31

-15

0

p1

39

-16

0

p1

51

-17

0

p1

61

-18

0

po

ol

N/A

N/A

N/A

Pre-vaccinationPost-vaccination

CD8 and CD4 epitopes

Enhanced multifunctional effector cells, and TCR avidity

3

2

1

0

d1699

preee

Multicytokine

Pre Mid End

EC

50

(p

ep

co

nc

. fo

r 5

0%

IF

N-g

+)

0.00001

0.0001

0.001

0.01

0.1

1

10

#001#003#004#005#006#007#010#011#012AvgGM-CSF

IFN

-g

100

101

102

103

104

100

101

102

103

104

7.75%48.27%

38.12%5.85%

IL-2

TN

F-a

100

101

102

103

104

100

101

102

103

104

43.48% 3.48%

48.70% 4.35%

100

101

102

103

104

100

101

102

103

104

47.38% 25.07%

20.73% 6.82%

IFN-g+ DP

DN GM-CSF+

100

101

102

103

104

100

101

102

103

104

1.56%93.97%

0.31%4.16%

100

101

102

103

104

100

101

102

103

104

19.35%56.77%

10.32%13.55%

Summary: I

• NY-ESO-1157-170 peptide; rV/rF induced integrated

humoral, CD4+ and CD8+ T cell responses in a

significant proportion of EOC patients.

• Improved survival in a phase II trial.

• ALVAC-NY-ESO-1/TRICOM vaccination elicits

NY-ESO-1-specific immune responses in all patients• multicytokine producing CD4 and CD8 T cells• Skews towards Th1polarization.• Generates T cells with higher TCR avidity compared

with pre-existing T cells.• Long lived functional CD4+ T and CD8+ T cells at 6

months in all patients, and at 12 months in some patients.

• OS 36-85 months

Overview of vaccination strategies in clinical trials

Vaccine Phase Tumor PTS

*

Note

Vaccines with viral vectors

PSA-TRICOM II Prostate 122 8.5 mos OS improvement

PANVAC-VF III Pancreatic 255 Failed >OS.

Vaccines with peptides

Sipuleucel-T (Provenge) III Prostate 512 4.1 mo OS improvement

Vitespen (Oncophage) III Melanoma 322 Prolonged OS in M1a or M1b

subpopulation

III Renal 818 No difference in DFS and OS

Vaccines with tumor cells or tumor–cell lysates

OncoVAX III Colon 254 Significant improvement in DFS

and OS in stage II

Reniale III Renal 558 Significant improvement in DFS

and OS

GVAX III Prostate 626 Failed to improve OS versus

docetaxel

Odunsi K, et al. PNAS. 2007; Nishikawa H et al., JI, 2006;

Qian et al, Cancer Res. 2009; Matsuzaki J. et al PNAS 2010.

Camouflage and sabotage: ovarian

cancer escapes from immune attack

• Role of regulatory T cells.

• Loss of antigen expression

• Role of molecules that

mediate immune tolerance

and T cell exhaustion:

• Indoleamine 2,3

dioxygenase (IDO)

• Program Death-1 (PD1)

• Lymphocyte Activation

Gene-3 (LAG-3)

• CTLA-4

APC

SHP-

2

P

PITSM

ITIM

PD-1

PD-1

ligand

P

Proximal

signaling

kinases

CD-3

TCR

MHC

Class II

LAG-3

KIE

ELE

KIE

ELE

P

Dephosphorylation

Less T

cell

activation

Cytoplasmic domain of

PD-1 contains two

motifs

• ITIM: Immunoreceptor

tyrosine-based

inhibitory motif

• ITSM:

Immunoreceptor

tyrosine-based switch

motif

PD-1 and LAG-3 co-inhibitory molecules

Expression of PD-1, LAG-3 and CTLA-4 on NY-ESO-1

specific CD8+ cells at the tumor site

5.2%

IFN-γ+ve

33.9%

IFN-γ+ve

The capacity for IFN-γ

production is diminished in

LAG-3+ and PD1+ subsets

of tumor-antigen-specific

T cells.

Clone RPOV10: HLACw3

Dual LAG-3 and PD-1 pathway blockade during priming efficiently

restores frequency and effector function of NY-ESO-1–specific

CD8+ T cells

Safety and Activity of PD1-PDL-1 pathway

blockade in humans

• Safety, activity, and immune correlates of

anti-PD-1 antibody in cancer. Topalian SL et al, N Engl

J Med; 366:2443, 2012.

- Cumulative response rates were 18% NSCLC, 28%

melanoma and 27% RCC; PD-L1-positive tumors (36%).

• Safety and activity of anti-PD-L1 antibody in

patients with advanced cancer. Brahmer JR, et al, N Engl

J Med; 366:2455, 2012.

- 9 of 52 patients with melanoma, 2 of 17 with renal-cell

cancer, 5 of 49 with non-small-cell lung cancer, and 1 of 17

with ovarian cancer.

TUMOR

CELLSIFN-γ

NK cell

CTL B cell CD-4+

T cellTreg MDSC TAM

Proinflammatory cytokines

(e.g. IL-6)

IDO-DC

Tumor

growth

Tumor

immunity

Critical questions:Which antigen(s)?

Vaccine / Immunotherapy strategies?-DCs Vs other platforms, ACT

Desirable immune response?- Memory Vs Effector

What are the most significant

mechanisms of immune evasion in

human cancer?

On-going and Future Work

Perspectives for 2012

• Consolidation with immune therapy to minimize

relapse is feasible, yields robust immunological

results.

• Combination of vaccination with strategies to

overcome immune suppression.

• Integration of cellular therapies (dendritic cells,

engineered T cells) into treatment paradigms.

AcknowledgementsRoswell Park Cancer Institute

Buffalo, NY

Ludwig Institute, NY

Ovarian Cancer

Research Fund

Junko Matsuzaki

Raya Huang

Feng Qian

Amy Beck

Tony Miliotto

Cheryl Eppolito

Shashikant Lele

Nefertiti DuPont

Sacha GnjaticErika RitterGerd RitterLinda PanRalph VenhausLloyd J. Old**

NCI R01CA158318-01A1

P30 CA016056-32