PHASE I VACCINATION TRIAL OF SYT-SSX JUNCTION PEPTIDE AND ITS HLA-A*2402 ANCHOR

SUBSTITUTE IN PATIENTS WITH DISSEMINATED SYNOVIAL SARCOMA

Kawaguchi S, Wada T, Nagoya S, Ida K, Sato Y,

Torigoe T, Sato N, Ishii T, Tatezaki S, Yamashita TSapporo Medical University

Chiba Cancer Center Hospital

Synovial sarcoma

Survival rate

Guillou L et al., J Clin Oncol, 2004

Peptidevaccination(MAGE-3)

Peptide immunotherapy for malignant melanoma

Coulie PG, Universite de Louvain , Brussels

Antigenic peptide

Tumor cell

T cell

T cell

T cell

T cell

T cellT cell

T cell

T cell

Dendritic cell

Patient

T cell

T cell

T cell

Injection

T cell

Peptide immunotherapy

Gene

T cell

ProteinAntigenic peptide

Tumor cell

Antigenic peptide

HLATCR

Activation

Desirable antigenic peptide

1.Tumor specific expression

2. High affinity to HLA

3. Immunogenicity

Desirable antigenic peptide

1.Tumor specific expression

2. High affinity to HLA

3. Immunogenicity

Tumor Fusion gene Frequency (%)

Synovial sarcoma SYT-SSX1 65 SYT-SSX2 35Ewing sarcoma EWS-FLI1 85

EWS-ERG 5-10Liposarcoma TLS-CHOP >95RMS PAX3-FKHR >75Clear cell sarcoma EWS-ATF1 >80DFSP COL1A1-PDGFb >99

Specific fusion gene

Desirable antigenic peptide

1.Tumor specific expression

2. High affinity to HLA

3. Immunogenicity

HLA-A24-binding motif

22 99 10101 3 456

7 8

2nd PortionY, F, W, or M

9th or 10th PortionF, L, I, W, R, or KHLA-A24

Tumor cell

T cell receptor

T cell

SYT SSX(1, 2)

…… …PQQRPYGYDQIMPKKPA EHAWTHRLRERK

Y, W, K, R: HLA-A24 binding motif

A: PYGYDQIMPK C: AWTHRLRER

B: GYDQIMPKK D: AWTHRLRERK

Peptides

SYT-SSX peptides

EBV NA24 F4.2 A B C D

1.8

1.6

1.4

1.2

1.0

0.8

0.6

0.4

0.2

0

Binding affinity to HLA-A24

SYT-SSX peptides

Desirable antigenic peptide

1.Tumor specific expression

2. High affinity to HLA

3. Immunogenicity

Reactivity to circulating T cells

◇◇

◇ ◇

T cell(CD8)

HLA/Peptide Tetramer

FACS

1.17%

HLA-A24-positiveSynovial sarcoma patients : 16 Other sarcoma patients : 10 Healthy individuals : 10

Synovial sarcoma

Other sarcomas

Healthy Individuals

-0.1

0

0.1

0.4

0.5

0.6

0.25

SYT-SSX A

SYT-SSX B

SYT-SSX C+D

Po

siti

ve c

ells

(%

)Reactivity to circulating T cells

Induction of cytotoxic T lymphocytes (CTL)

Peptide stimulation(SYT-SSX B)

Mixed CultureCr release assay51

T cells

Synovial sarcomaCell line

HLA-A24 SYT-SSX●Fuji (+) (+)

○HS-SY-II (+) (+)

■SW982-A24 (+) ()□K562 () ()

E/T ratio

30

20

10

0

40

30 10 3

%S

pec

ific

Lys

isCTL induction

Sato et al., J Immunol, 2002

SYT-SSX B peptide

1.Tumor specific expression

2. High affinity to HLA-A24

3. Immunogenicity

SYT-SSX

SYT SSX

T cellHLA-A24

SYT-SSX B

Rationale

・・ PQQRPYGYDQIMPKKPA・・・

SYT-SSX B peptide

Synovial sarcoma cell

T cell

T cell

T cell

T cell

T cellT cell

T cell

T cell

Dendritic cell

Patient

T cell

T cell

T cell

Injection

T cell

Phase I clinical trial

To evaluate

(i) Toxicity

(ii) Immunological property

(iii) Anti-tumor effect

of SYT-SSX B peptide vaccine in patients with disseminated synovial sarcoma

Purpose

Eligibility1. Histologically diagnosed synovial sarcoma

2. SYT-SSX positive

3. HLA-A*2402 positive

4. Unresectable tumors

5. One month or more after chemotherapy

Protocol

2 108640

Peptide: SYT-SSX B: GYDQIMPKK

Schedule: Every two weeks

Dose: 　 0.1mg in 3 patients

　　　　 1.0mg in 3 patients 　　　

1. Toxicity 　 National Cancer Institute of Common Toxicity Criteria

2. Immunological property(i) Delayed-type hypersensitivity (DTH)(ii) HLA-A*2402/peptide tetramer(iii) In vitro CTL induction

3. Anti-tumor effectCT scan

Evaluation

Results

Case Age Gender Dose No. of immunization

1 69 M 0.1mg 1

2 32 M 0.1mg 3

3 21 F 0.1mg 6

4 21 M 1mg 6

5 39 F 1mg 6

Participants

6 26 M 1mg 4

Toxicity DTH

－ －－ －－ －－ －

Fever(Grade1)

－

Toxicity and DTH

Case

12345

－ －6

Before vaccination After 1st vac. After 6th vac.

HIV

te

tram

er B

tet

ram

er 0.06 0.41 0.47

0.01 0.00 0.01

Tetramer analysis (Case 4)

Case Pre-vac. 1st vac. 　 3rd vac. 　 6th vac.

2 0.02 0.02 3.05 N.D3 0.42 0.49 0.52 0.624 0.06 0.41 0.36 0.475 0.50 0.52 0.09 0.036 0.02 0.15 0.08 N.D

Tetramer analysis

Case Pre-vac. 1st vac. 　 3rd vac. 　 6th vac.

2 Failure Failure Success ND3 Success Success Success Failure4 Failure Success Success ND5 Failure Success Failure Failure6 Failure Failure Failure ND

CTL induction

May 15 June 18

Anti-tumor effect (Case 3)

Anti-tumor effect (Case 5)

July 8 July 22

Sept 16Aug 19

Discussion

Clinical trials of fusion-gene peptides

Tumor Peptide Adjuvant Remission

SS Class I None 0/6

ES Class I IL-2 (9x106IU/m2) 1/12

RMS Class I IL-2 (9x106IU/m2) 0/4

CML Class I QS-21 5/14Class II

Modification of peptide and protocol

1. Anchor motif substitution

2. Adjuvant and cytokine

3. Class II peptides

4. Protocol at the adjuvant setting

Tumor

G D Q I M P K KY

HLA-A24

Anchor motif substitution

SYT-SSX B peptide

W （ tryptophan

）

L （ leucine ）

F （ phenylalanine ）I （ isoleucine ）

0

10

20

30

40

%S

pe

cif

ic L

ys

is

Fuji HS-SYII SW982 K562

0

10

20

30

40

E/T=30

E/T=10

E/T=3

Fuji HS-SYII SW982 K562

B peptide K9I peptide

CTL induction

Ida et al., J Immunol, 2004

Before vaccination After 1st vaccination

Vaccination of K9I peptideH

IV

tetr

amer

B t

etra

mer 0.02

0.00

0.41

0.01

1. The safety and efficacy of SYT-SSX B peptide were evaluated in 6 patients with disseminated synovial sarcoma.

2. A total of 16 vaccinations were carried out, resulting in (i) no serious adverse effects or DTH reactions, (ii) tumor dormancy in 1, (iii) increases in CTL in 3, and (iv) CTL induction from 4 patients.

3. The present trial demonstrated the safety and immunogenicity of the peptide. Subsequent trial using a modified peptide and adjuvants is currently underway.

Conclusion