1

Adoptive T-cell therapy

Translation of clinical efficacy in melanoma

to pancreatic cancer

Rienk Offringa

K.H. Bauer foundation endowed professor

Div. Molecular Oncology of Gastrointestinal Tumors, DKFZ

Div. Pancreas Cancer Research, Uniklinikum Heidelberg

2

‘Tumor-observing’ T-cell response in human

melanoma

T-cell

OFF ON

Dominance of

inhibitory signals

Tumor Infiltrating Lymphocytes (TILs)

3

Mobilization anti-tumor T-cell response

Go!

T-cell

OFF ON

Tumor Infiltrating Lymphocytes (TILs) Block

inhibitory signals

Cancer vaccines

Pre-requisites:

• Time (> 3 months)

• Immune system in good shape

• 1st (2nd?) line treatment

4 Adoptive T-cell therapy for

advanced, metastatic melanoma

ex vivo expansion

re-infusion tumor biopsy

TIL

TIL

Dudley/Rosenberg

OR rate: ~50%

Durable CR: ~20%

Advanced melanoma

> 300 patients

Also in European centers (but not yet in Germany)

TIL isolation

TIL = tumor-infiltrating lymphocyte

5

Proof of concept for TIL therapy in Heidelberg

ex vivo expansion

re-infusion tumor biopsy

TIL

TIL

TIL isolation

TIL = tumor-infiltrating lymphocyte

Clinical trial

• Metastatic melanoma (n=8)

• Progression after initial response to

checkpoint inhibitors (CTLA-4; PD-1)

• TIL-infusion & PD-1 blockade

Primary endpoint:

• Clinical response (RECIST)

Secondary endpoints:

• Toxicity

• Longevity TIL response (TCR deep seq.)

• Molec. tumor imaging

Further biomarkers:

• Antigen-specificity TIL response

• Tumor-antigen expression

A. Enk, D. Jäger, R. Offringa

6

Proof of concept for TIL therapy in Heidelberg

ex vivo expansion

re-infusion tumor biopsy

TIL

TIL

TIL isolation

TIL = tumor-infiltrating lymphocyte

Objectives:

• Offer TIL therapy to melanoma patients

in Heidelberg/Germany

• Combine with other approaches

(checkpoint blockade, vaccines, other)

• Prelude to studies with engineered T-

cells (TCR, CAR)

• Prelude to T-cell therapy in other

cancers

7

4 4

UK: 5th

USA: 4th

Pancreatic cancer-related death: 4th

(Germany 2012)

8

GER

UK

USA

Incidence*

11.5

9.6

10.9

Death*

11.1

9.0

10.9

Lifetime risk

~ 1.4%

* yearly cases/100.000

Pancreatic cancer

Incidence and death rates almost equal

9

Trends in 5-year survival rates (U.K.)

GER: 7%

USA: 6% age-standardized rates

Overall survival rates barely improved in 40 years (unmet medical need)

10

Pancreas cancer management to date

• Surgery only effective treatment option

• Fraction of eligible patients: 20-30%

• Devastating recurrence rate

• Chemotherapy: adjuvant treatment & palliative care

Markus Büchler

Oettle JAMA 2007 70% recurrence

in 2 years!

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T-cell therapy for countering recurrence

Oettle JAMA 2007 70% recurrence

in 2 years!

Isolate TILs from operated patients freeze

2. TIL therapy as adjuvant treatment

(4 wks needed for preparing TILs)

1. TIL therapy at tumor recurrence

(palliative setting)

12 Pancreatic cancer has been branded

poorly immunogenic

• Highly malignant

• No spontaneous regressions

• Very few T-cells

• T-cells cannot be isolated and cultured

Don Quichote Is immunotherapy of pancreatic cancer überhaupt a valid concept?

13

T-cell infiltrate found in majority of PDA samples

450 CD3+/mm2

280 CD3+/mm2

48 CD3+/mm2

Low (<100) Medium (100-300) High (>300)

~20% ~ 20% ~ 60%

CD3+ T-cells

14 Ex vivo expansion of Tumor-Infiltrating Lymphocytes

the melanoma TIL protocol

tumor

activated T cells

cluster of

proliferating T cells

Minced tumor bits

in 24 well format

with high-dose IL-2

Rapid expansion on

allogeneic feeders,

anti-CD3 and IL-2

Large number of

activated T cells

2 w

eek

s

2 w

eek

s

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A lot of experiments

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Mel008 TOP50 tumor

blood tumor tumor unique0

1

2

3

4

10

20

rela

tive

fre

qu

en

cy (b

eta

-se

q)

36/50

blood tumor tumor unique0.0

0.2

0.4

0.6

0.8

1.01.01.52.02.53.0

Tumor

enriched

Tumor

unique

Tumor

enriched

Tumor

unique

Pancreatic cancer Melanoma

Poschke, Hassel, Volkmar, Sahin & Offringa

TC

R-b

eta

re

lative

fre

que

ncy (

%)

TCR-seq: evidence for tumor-reactive TIL clones

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IHC: Tertiary Lymphoid Structures (TLS)

CD3 CD20

CD208 (DC-Lamp+ DC) CD21 (follicular DC)

• TLS in approx. 50% of PDA biopsies

• Definition: B and T cell areas with

appropriate DC subsets; HEV

• Associated with good prognosis in

lung, breast and colorectal cancer

Poschke, Halama, Bergmann & Offringa

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All TCR+ T-cells (CD3)

V-beta 2+ T-cells

Poschke, Halama, Bergmann & Offringa

Tertiary

Lymphoid

Structures

Evidence for clonal expansion in TLS

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Planning 2015-2016

Perform TIL study in melanoma

• GMP test runs

• Trial

Pre-clinical proof of concept for tumor-reactivity TILs in pancreatic cancer

• Cloning TCRs from TILs

• Tumor-reactivity in vitro

• Antigen-specificity (exome data)

• In vivo xenograft experiments

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Acknowledgements

European Pancreas Center

Markus Büchler

Oliver Strobel

Thilo Hackert

Christine Tjaden

Nathalia Giese

Pathology

Frank Bergmann

Offringa-lab DKFZ

Isabel Poschke

Michael Volkmar

Jenny Hermes

Janina Rebmann

Claudia Lauenstein

DKFZ

Andreas Trumpp

Martin Sprick

Roland Eils

Matthias Schlesner

Thomas Höffner

Michael Floβdorf

Michael Platten

NCT

Niels Halama

Dirk Jäger

Jessica Hassel

Alexander Enk

BioNTech

Ugur Sahin

Marta Faryna

Martin Loewer

GMP

Stefan Eichmüller

Michael Schmitt

Anthony Ho

Stefan Meuer

21 Melanoma TILs target mutated self antigens

hypothesis: same applies to PDA TILs