Optimal-affinity T cell receptors

from humanized TCR mice Elisa Kieback, May 2017

Immunotherapy for Cancer

Cancer-specific receptors

TCR gene therapy

Cancer patient

Cancer-specific T cell

Cancer-specific receptor

T cell

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CASH

BILLS

PROBLEM

NOT EFFICIENT STRATEGY NOT SAFE DMF4

DMF5 CEA

p53

MAGE-A3 MAGE-A3a3a

DMF5

???

HBV

gp100

MAGE-A10

Phase I/IIa

Phase IIb

CARs TCRs

•  CD19 CAR T cells: success story for leukemia/lymphoma

•  Up to 90% complete remission in end-stage patients

•  Target only cell surface antigens

•  „Breakthrough Therapy“ designation by FDA

•  Employ natural T cell signaling •  Target also intracellular

antigens •  Target microenvironment •  More efficient for solid tumors

How do we find TCRs that are safe and efficient?

MAGE-A4

1D3 WT1

HPV E7 HPV E6

Tyr

gp100

mNY-ESO

NY-ESOc259

Technology – Humanized TCR Mouse

yHuTRA (~1400 kb)

Vα1...41 Cα Jα

yHuTRB (~700 kb) Jβ1 Jβ2

Cβ1 Cβ2 Vβ2...30 D1 D2

Human TCR gene loci transgenic mice

HLA-A*02:01 HLA-DR4

IFN-g

Immunization of mice

Mice are optimal TCR factories

•  Most tumor-associated antigens (TAAs) are self-proteins

•  Deletional tolerance of high avidity T cells in humans

•  Many TAAs differ between mouse and human

•  More efficient immune response can be elicited in vivo

à BUT: Mouse TCRs are immunogenic in humans

antigen

Characterize immune response in blood

Isolate reactive T clells

MAGE-A1278 a – fetoprotein158 MAGE-A10254 NY-eso157

4.7 0.9 1.1 3.0

CD

8

p/MHC multimer

5‘ LTR TCRβ 3‘ LTR P2A TCRα WPRE Ψ"

Competing technology Efficacy (TCR affinity)

Safety (no off-target toxicity)

Technology (feasibility/robustness)

Humanized TCR mouse

✔Optimal affinity

✔Self-restricted

"

✔Efficient priming TCR selection

Autologous T cells/TILs" ✖ " ✔ ✖ "Allo-restricted T cells" ✔ ✖ " ✖ "Affinity enhancement" ✔ ✖ " ✖ "HLA-transgenic mice" ✔ ✖ " ✔

Differentiation

2/5 mice 8/10 antigens

Highly robust immune response

Example: MAGE-A1 TCR

•  Multiple myeloma •  Lung cancer (NSCLC) •  Melanoma •  Breast cancer •  Colon cancer •  Hepatocellular

carcinoma •  ... 0

20

40

% M

AG

E-A

1+ bi

opsi

essingle cells1-5%10-30%40-60%80-100%

13%

MAGE-A1+ plasma cells

(n=137)

Efficacy

RP

MI8

226

OP

M-2

NC

I-H92

9

RP

MI8

226

OP

M-2

NC

I-H92

9

U26

6

L363

0

1000

2000

IFN

g (p

g/m

l)

MAGE-A1

HLA-A*02

- + - + +

+ - + + +

+ + + + + + + +

- - - + + + + +

Myeloma Melanoma/Lung

Recognition of tumor cells Rejection of tumors in mice

b a

TRIM

25

SA

MD

9

CO

L28A

1

MM

P27

DA

PK

3

MA

GE

-A1

PM

A/io

no

T2 a

lone

IFN

-γ (p

g/m

l)

Donor 1 Donor 2

Sequence Gene IC50 (nM)

X X X E Y X I K X T1376 motif

K V L E Y V I K V MAGE-A1 4.11

E L L E Y Y I K V TRIM25 23.34

G L L E Y L I K S SAMD9 73.23

K Q F E Y D I K T MMP27 834.74

R S L E Y D I K L COL28A1 4258.75

R L K E Y T I K S DAPK3 5212.32

Potential off-target epitopes

No relevant off-target toxicity

No recognition of: •  Allo-MHCs •  Primary cells of vital organs

(lung, heart, bone marrow) •  Broad panel of self-peptides

-13 -12 -11 -10 -9 -8 -7 -6 -5 -40

50

100

peptide concentration (log M)

IFNγ

(% o

f max

.)Donor 1Donor 2Donor 3

Donor 1Donor 2Donor 3

SAMD9

MAGE-A1

No expression in normal tissue

Safety

à Initiation of Phase I/IIa trial

Antigen expression

MAGE-A1+ HLA-A2+

Commercial Potential

MAGE-A1 TCR / multiple myeloma (US per year, 2016)

New cases 30,330 Deaths 12,650

HLA-A2+ 6325 MAGE-A1+ 822

Costs per treatment 300,000 $ Market penetration 15%

Estimated sales 37 Mio $

Application for many indications!

Ø  Cancer Immunotherapy Market (Bio US$, Markets & Markets 2017)

Ø  Multiple Myeloma Market (Bio US$, Global Data 2017)

2016 2021

61.9 119.4

2016 2023

12.8 22.6

Preclinic IND Phase I/IIa Discovery

Development Status

NY-ESO (CD4+CD8)

HPV16 E6

Neoantigens (>20)

New HLA variants

A*03:01

A*11:01 C*04:01 B*15:01

B*07:02 C*07:02

MAGE-A1

End of 2017

> 100 antigens

analyzed in vivo

> 500 mice

immunized

> 250 TCRs cloned

> 10 billion T cells

screened

DR1

DR7 DR15

DR3

MHC I MHC II

TCR IP package

Transfer system

Humanized TCR mice

Individual TCRs Groups of TCRs Safety switch

FTO with retroviral vector Patent on Transposon technology

Licence agreement with MDC spin-off

Intellectual Property / Commercialization

What we are looking for 1. VC investment for (pre-) clinical product

development

2. Biotech partner for clinical development of

TCR candidates and sublicensing

3. Fee-for-service TCR isolation

GMP production Validated process Manufacturing licence pending

Complex Know-how No patent filed FTO

Thank you. Max Delbrück Center (MDC)

Thomas Blankenstein Gerald Willimsky Liang-Ping Li Vivian Scheuplein Stephanie Petzold Ioannis Gavvovidis Lucia Poncette Xioajing Chen Dana Hoser Christoph Lampert Arunraj Dahmodaran Meng-Tung Hsu Georgios Papafotiou Wolfgang Uckert Matthias Leisegang

Charité

Antonio Pezzutto Matthias Obenaus Igor Blau Axel Nogai

Holger Specht

GMP facility

Joachim Kopp Martin Vaegler Hana Rauschenbach

Stem Cell Facility, Charité

Charité Research Organization

Medigene AG

Dolores Schendel Carina Wehner Sabine Hauck Christine Leichtle Christiane Eck Silke Raffegerst

S F B – T R 3 6