4/8/2016
The Immunology and Genetics of
Cancer Immunotherapy
ACCME/Disclosure
Dr. Yang has nothing to disclose
James Yang
Surgery Branch, NCI
March 13, 2016
Adoptive Cellular Transfer:
T-Cells as Therapy
Melanoma TIL in IL-2
(Tumor Infiltrating Lymphocytes)
• Adoptive cell therapy (ACT) with
tumor reactive T-cells can cause
complete and durable tumor rejection
in some patients with advanced cancer
• This was first optimized and then
demonstrated in patients with
metastatic melanoma using tumor
infiltrating lymphocytes (TIL)
Fresh digest
One week
Two weeks
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T-Cell Adoptive Therapy
• Transferring tumor-reactive T-cells
activated and expanded in vitro rapidly
establishes an anti-tumor repertoire
• Ex-vivo expansion permits the use of
reagents and methods not tolerated in
vivo
• This also allows the separate
manipulation of reactive T-cells and
the inhibitory tumor microenvironment
to optimize efficacy
Preparative Host
Immunosuppression
• Immunosuppressing the recipient prior
to T-cell transfer enhances T-cell
survival and efficacy
– Removes resident Tregs
– Induces homeostatic cytokines
– Reduces competition for cytokines (‘sinks’)
– Non-specifically increases TLR ligands (LPS)
Cyclophosphamide + Fludarabine
Non-Myeloablative Chemotherapy
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CR Metastatic Melanoma
TIL for Metastatic Melanoma
• Between 2000 and 2007, 93 patients with
measurable metastatic melanoma were
treated with a preparative lymphodepleting
regimen followed by TIL and IL-2
Overall RR= 56%
PR= 34%
CR= 22%
• 86% had visceral metastases
• 83% had tried prior IL-2
• Only two patients had a second treatment
12
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Melanoma TIL Protocol #2
• Beginning in 2011, 101 additional pts
received TIL and IL-2 after being
randomized to high or low intensity
lymphodepletion (p= N.S.)
• 3.4 yr median FU
• One treatment related mortality
• Overall RR for all pts is 54% with
24% CRs (only one CR has relapsed)
Metastatic Melanoma‐ CR
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TIL Responses in Melanoma Patients Refractory to Other Immunotherapies
N= 194
ORR= 55%
PR = 35%
CR = 20%
What Are the Tumor-Associated
Antigens Being Recognized?
• Tissue differentiation antigens (MART1,
gp100)
• Tumor germline antigens (NY-ESO1, MAGE)
• Normal proteins overexpressed by cancer
cells (hTERT, EGFR)
• Proteins containing tumor specific mutations
(MUM-1, CDK4, B-catenin, erbB2IP)
Prior
Immunotherap
y
N=
ORR
(%)
High Dose IL-2
29
Anti-CTLA4
only
CR
(n=)
PR
(n=)
59%
8
9
31
61%
9
10
Anti-PD1 only
3
66%
1
1
Anti-CTLA4
and
Anti-PD1
8
25%
1
1
Categories of Tumor Associated Antigens
• Tissue differentiation antigens
• Tumor germline antigens
• Normal proteins overexpressed by
cancer cells
• Proteins containing tumor specific
mutations
• Viral proteins
• Viral proteins (HPV, EBV, MCC)
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Categories of Tumor Associated Antigens
• Tissue differentiation antigens
• Tumor germline antigens
• Normal proteins overexpressed by
cancer cells
• Proteins containing tumor specific
mutations
• Viral proteins
Targeting Shared Antigens with
Receptor-Engineered T-Cells
Unmutated Self
Antigens
• Constant between
patients (off-theshelf reagents)
• Potential for
autoimmune toxicity
• T-cell repertoire
may be limited by
thymic deletion
Mutated Non-Self
Antigens
• Very patient
specific
• Very low potential
for autoimmunity
• No central thymic
tolerance
(‘neoantigens’)
TCRs & Chimeric Antigen
Receptors (CAR)
• High-efficiency, stable gene insertion into
mature human T-cells is possible using viral
gene-therapy vectors
• “One-shot” replication-incompetent
retroviruses have been safely used to modify
T-cells in hundreds of patients
• Native alpha-beta T-cell receptors (TCR) as
well as chimeric antigen receptors (CAR)
have been used
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Adoptive Transfer with Receptor
Engineered T-Cells
Targeting Melanocytic Proteins:
Anti-MART1 TCR-Engineered PBL
• Tissue differentiation antigens (MART1,
gp100, CEA, CD19)
• Tumor germline antigens (NY-ESO1, MAGE)
• Normal proteins overexpressed by cancer
cells (hTERT, EGFR)
• Proteins containing tumor specific mutations
(MUM-1, CDK4, B-catenin, erbB2IP)
• Viral proteins (HPV, EBV, MCC)
PBL with TCR Targeting CEA
Successful Targeting of Normal Ags
• Tissue differentiation antigens (MART1,
gp100, CEA, CD19)
• Tumor germline antigens (NY-ESO1, MAGE)
• Normal proteins overexpressed by cancer
cells (hTERT, EGFR, mesothelin)
Effective attack on normal self-antigens
may cause unacceptable autoimmunity
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Anti‐CD19 CAR: Follicular Lymphoma
Primary Mediastinal B-Cell Lymphoma
Successful Targeting of Normal Ags
Gene Therapy with Anti-NY ESO-1 TCR
(Melanoma)
• Tissue differentiation antigens (MART1,
gp100, CEA, CD19)
• Tumor germline antigens (NY-ESO1, MAGE)
• Normal proteins overexpressed by cancer
cells (hTERT, EGFR, mesothelin)
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Synovial Sarcoma
What Are the Native Antigens
for TIL?
• When patients respond to TIL,
autoimmune sequelae are rare
• T-cells against many of the
described antigens (such as the
tumor-germline Ags) are not
found in most therapeutic TIL
Categories of Tumor Associated Antigens
• Tissue differentiation antigens (MART1,
gp100, CEA, CD19)
• Tumor germline antigens (NY-ESO1, MAGE)
• Normal proteins overexpressed by cancer
cells (hTERT, EGFR)
• Proteins containing tumor specific mutations
(MUM-1, CDK4, B-catenin, erbB2IP)
• Viral proteins (HPV, EBV, MCC)
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T‐Cells Can Recognize Tumor‐Associated Mutated Antigens
Somatic mutation frequencies observed in exomes
from 3,083 tumour–normal pairs.
J Ex Med, 1996
MS Lawrence et al. Nature 1-5 (2013) doi:10.1038/nature12213
How Often Do Melanoma TIL Recognize
Tumor-Specific Mutated Antigens?
• Perform whole exomic sequencing (WES)
on melanomas containing tumor-reactive
TIL and identify mutations
• In silico, list every 9-mer or 10-mer
epitope that contains a mutated amino
acid (19 candidates per mutation) in
order of predicted binding to the MHC
allele of the patient
• Test top binders for TIL recognition
Exomic sequencing of melanoma 2369
(570 mutations; 10830 Candidate Peptides)
Rank
Peptide
Mutation
Affinity
(nM)
Gene
IFN- (pg/ml)
<30
10400
1 FSDYYDLSY
2 LTDDRLFTCY
117 G to S
1005 H to Y
2
3
C22orf33
PLEKHM2
3 YSSALDLCY
621 N to D
5
GRIN3B
<30
4 FSDKKVGTY
688 L to F
5
PLCB1
<30
5 HSEYSSFFY
603 H to Y
6
HEG1
<30
6 CSNFLLLAY
84 S to L
7
BAI3
<30
7 ESDKEELVGY
332 F to L
7
MPP4
<30
8 CTDTYMLELF
191 H to Y
8
OR4C46
<30
9 FTGTISVMY
60 P to S
12
UEVLD
<30
10 QTQSVVFLY
156 S to L
13
COL9A1
<30
11 MSSYIASFTY
356 L to F
14
LST-3TM12
<30
12 CTDTYMLEL
191 H to Y
22
OR4C46
<30
13 LLDLMAYDRY
117 G to D
22
OR2T2
<30
14 SSDSQEENY
117 G to E
23
MEOX2
<30
15 LTSMAYDCY
122 R to C
31
OR8B3
16 YTDFHCQYV
176 P to H
49
PPP1R3B
17 WADWGHRTY
3344 A to T
51
LRP2
<30
18 FTMVILYVVY
219 S to L
54
LRRC3B
<30
19 CVDSPPPLFF
528 S to F
71
C15orf2
<30
20 VSDGFTAVM
198 P to S
85
RNPEP
<30
<30
13400
Robbins et al, Nat Med 2013
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Mismatch Repair and Response to
Pembrolizumab
Mismatch
Mismatch
Repair
Repair
Proficient CRC Deficient CRC
Mismatch
Repair
Deficient
Non-CRC
Pts Treated
18
10
7
Complete
Responses
0
0
1
Partial Responses
0
4
4
Response Rate
0%
40%
71%
Pt tumors
sequenced
6
7
2
73
1875
1455
Mean # somatic
mutations
Le et al, NEJM 2015
Science, May 2014
Tumor Burden
(Sum of Max Lesion Diameters)
43 yo F with cholangiocarcinoma to liver and lungs
Lung Relapse
Treated with bulk unselected TIL ‐ minimal response
Identified TIL culture reactive with one of her 26 tumor mutations (in mut‐ERBB2 interacting protein)
This culture expanded and pt was treated identically but with this selected culture
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Treatment #2: Selected TIL
Treatment #1: Unselected TIL
45
Unmutated Self
Antigens
• Constant between
patients (off-theshelf reagents)
• Potential for
autoimmune toxicity
• T-cell repertoire
may be limited by
thymic deletion
Mutated Non-Self
Antigens
• Totally patient
specific
• Very low potential
for autoimmunity
• No central thymic
tolerance
(‘neoantigens’)
Shared Mutated
Antigens
• Constant between
patients (off-theshelf reagents)
• Very low potential
for autoimmunity
• No central thymic
tolerance
(‘neoantigens’)
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RAS Pathway
The KRAS gene is the second-most mutated gene in human
cancer (second only to p53). Approximately 25% of human
cancers have abnormalities in KRAS.
Identification of T-Cell Receptors Targeting
KRAS-Mutated Human Tumors
ACT of Colon Cancer with Anti‐G12D KRAS TIL
(HLA‐C*08:02 Restricted)
Qiong J. Wang*, Zhiya Yu, Kayla Griffith, Ken-ichi Hanada,
Nicolas P. Restifo, James C. Yang*
Published online Dec 23, 2015, Cancer Immunol Res
Two high avidity HLA-A*1101 restricted T-cell receptors
that recognize G12V and G12D mutated RAS proteins were
generated by immunizing mice engineered with HLA-A11
PBL transduced with these murine receptors recognize
HLA-A11 tumor lines with the appropriate mutations
HLA-A*1101 is in approx 13% of the U.S. population
but is the most common Class I allele in Han Chinese
Tran et al,
Science Dec 2015
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Conclusions
Conclusions
• Melanoma has taught us that T-cell
adoptive transfer can induce complete
and durable tumor rejection in some
patients
• T-cell therapy relies on finding
safe antigens to attack
• The native antigens recognized by
melanoma TIL are often tumorspecific mutated neoantigens
• Personalized ACT will require
coordination with pathologists to
characterize a patient’s tumor
antigen profile
• Receptor gene engineering of T-cells
and selection of TIL for this reactivity
could allow application to any cancer
TIL Protocols
•
•
•
•
•
•
Cutaneous melanoma
Non‐cervical HPV‐related cancers
Non‐small cell lung cancer*
Common GI adenocarcinomas*
Ovarian cancer*
Ocular melanoma
* Mutation-reactive selected TIL (2-3 months)
Gene Therapy T-Cell Protocols
Target
TCR/
CAR
HLA Tumors
MAGE‐A3
TCR
A1 or Melanoma, various epithelial DP04
NY‐ESO‐1
TCR
A2
Melanoma, synovial sarcoma
EGFR v.3
CAR
‐‐
Glioblastoma
Mesothelin
CAR
‐‐
Ovarian, mesothelioma, ?pancreas
Thyroglob
TCR
A2
Differentiated thyroid cancer
HPV 16 E6
TCR
A2
Any HPV 16 cancer
CD70 (pend) CAR
‐‐
Clear cell renal, thymic carcinoma/thymoma
RAS (pend)
G12V/G12D
A11
Any G12V or G12D RAS mutated cancer
TCR
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Acknowledgements:
Steven Rosenberg
Chief, Surgery Branch, NCI
Paul Robbins
Stephanie Goff
Qiong Wang
Kenichi Hanada
Rob Somerville
– TIL Lab
• Steve Feldman
• Eric Tran
•
•
•
•
•
•
•
•
•
Yong-Chen Lu
Todd Prickett
Mark Raffeld
Richard Lee
• Immunotherapy Staff
• Clinical Fellows and
Nursing Staff
15