combination opportunities in immunotherapy with ... · hospital universitario reina sofía,...

Miguel F Sanmamed

María Rodriguez

Carlos Alfaro

Carmen Oñate

Inmaculada Rodriguez

José L Pérez Gracia

Bruno Sangro

Salvador Martin -Algarra

JM Lopez Picazo

Alfonso Gúrpide

Javier Rodriguez

Jesús San Miguel

Jesús Prieto

Alberto Benito

Ivan Peñuelas

Alvaro Gonzalez

Mercedes Iñarrairaegui

Aizea Morales

Arantza Azpilikueta

Elixabet Bolaños

Inmaculada Rodriguez

Sara Labiano

Eneko Elizalde

Alfonso Rodriguez

Angela Aznar

Maria Rodriguez-Ruiz

Pedro Berraondo

Luna Carneiro

Ines Guetgeman

Juan M Zapata

Manuel Rodriguez

Valerie Lang

Bettina Weigelin

Peter Friedl

Bristol Myers Squibb

Roche/Genentech

Pfizer

AstraZeneca

Boehringer Ingelheim

IGNORANT T CELL

Insufficient antigen/

lack of co-stimulatory signals

ANERGIC T CELL

Unsuccessful stimulation/

negative regulation

APOPTOTIC T CELL

(DELETED)

Programmed cell death

NAIVE T CELL PRIMED T CELL EFFECTOR T CELL MEMORY

T CELL:

Central memory cell

Effector memory cellCD27

CD28

HVEM

TCR

MHC

-peptide

CD27L

CD80/86

LIGHT

CTLA-4

B7-H4R

CD80/86

B7-H4

PD-1

B7-H4R

B7-H1

BTLA-4

B7-H1

B7-DC

B7-H4

BTLA-4R

CD137

OX40

ICOS

B7-H3R

CD137L

OX40L

B7h

B7-H3

IL-15

IL-7

CD-137

OX-40

others

IL-12

IFN-α

Schematic representation of the concept of immunostimulatory mAbs.

Melero I et al. Clin Cancer Res 2013;19:997-1008

©2013 by American Association for Cancer Research

Immune Checkpoint Blockade

against Malignancy

Blocking

Self-inflicted

Inhibitions?

CD80

CD86

CD28

CTLA-4

+

-

Anti-CTLA-4

Phase III clinical trials

Registered for metastatic melanoma.

Combines with vaccination preclinically.

10-20% sustained responses in phase II

30% incidence of moderate/severe autoimmunity

CTLA-4-/- dies because of multiorgan

lymphocyte infiltration

Response to Ipilimumab 10 mg/kg x 2 doses

No progression 5+ years

Survival Rate Ipilimumab + gp100 Ipilimumab alone gp100 alone

1-yr 44% 46% 25%

2-yr 22% 24% 14%

Hepatocellular carcinoma

A major target for immunotherapy

with checkpoint inhibitors.

Killing two birds with one stone

Phase II Trial of Anti-CTLA-4 Human Monoclonal

Antibody CP-675,206 in Patients With Advanced

HCC

Clinica Universidad de Navarra,

Pamplona

Hospital Universitario Reina Sofía,

Córdoba

Hospital Universitario 12 de Octubre,

Madrid

Protocol code CT-2007-01

EudraCT no. 2008-001177-15

NCT Identifier NCT01008358

Sponsor: University of Navarra, Pamplona, Spain

• Tremelimumab (CP 675,206) is a fullyhumanized IgG2 mAb that antagonizesbinding of CTLA-4 to B7 ligands.

– Antitumor activity has been demonstrated in a variety of murine tumor models

– It has shown efficacy on melanoma patients after single ormultiple doses. Similar drug Ipilimumab already approvedfor metastatic melanoma.

– It had not been tested against hepatocellular carcinoma

– Its antiviral effect had not been tested whatsoever

Tumor Response (Efficacy Population)

• Median number of courses: 2 (range: 1-4)

– At least 2 courses: 13 (76%)

– At least 4 courses: 6 (35%)

3 10 4

0% 20% 40% 60% 80% 100%

Disease Control Rate: 76.4%

Stable disease ≥ 6m: 3 (30%)17%

Tumor Response on Target

Lesions

-80

-60

-40

-20

0

20

40

60

Antiviral EfficacySerum HCV Viral Load

1,00E+00

1,00E+01

1,00E+02

1,00E+03

1,00E+04

1,00E+05

1,00E+06

1,00E+07

1,00E+08

0 30 120 210 300 360 390 530

Co

pie

s/m

L

Days after initiation of therapy

Patients with virological follow-up longer than 1 month

rCore

0 30 120 210 300 3300

10

20

30

40

50

Days after treatment

SF

C

rNS3

0 30 120 210 300 3300

20

40

60

Days after treatmentS

FC

rNS4

0 30 120 210 300 3300

50

100

150


SF

C

rNS5

0 30 120 210 300 3300

20

40

60

80


SF

C

pepP7

0 30 120 210 300 3300

10

20

30

40

50


SF

C

pepE2

0 30 120 210 300 3300

50

100

150


SF

C

pepE1

0 30 120 210 300 3300

50

100

150


SF

C

pepCore

0 30 120 210 300 3300

20

40

60

80


SF

C

pepNS5

0 30 120 210 300 3300

100

200

300

400

500


SF

C

pepNS4

0 30 120 210 300 3300

50

100

150


SF

C

pepNS3

0 30 120 210 300 3300

50

100

150

200


SF

C

pepNS2

0 30 120 210 300 3300

20

40

60

80


SF

C

*

*

*

Immunological Outcomes

Global Changes in Anti-HCV Immune Response

Immune Checkpoint Blockade

against Malignancy

Blocking

Self-inflicted

Inhibitions?

PD-L1 and IDO-1 permit placentation across MHC barriers

Why might PD-L1 be important?

Anti-tumour immune

responses are turned off

Promoting anti-tumour immune

responses

T cell activationT cell inactivation

PD-1

PD-L1/PD-L2

TCR

MHC

PD-1TCR

MHC No PD-L1

Partner cell

T cell T cell

Partner cell

Why might PD-L1 be important?

Anti-tumour immune

responses are turned off

Mechanism to promote anti-tumour

immune responses

T cell activationT cell inactivation

PD-1

PD-L1/PD-L2

TCR

MHC

PD-1TCR

MHCAnti-PD1

PD-L1

Partner cell

T cell T cell

Partner cell

PD-L1 blockade PD-1 blockade

PD-1 blockade = PD-L1 blockade?

Keir, et al. Annu Rev Immunol 2008

TCRMHC

PD-L1

PD-L2

B7.1PD-L1

B7.1

PD-1

-

APC T cell

TCRMHC

PD-L1

PD-L2

B7.1PD-L1

B7.1

PD-1

-

APC T cell

-

PD-1/PD-L1 blockade: a simple

paradigm

MHC=major histocompatibility complex; TCR=T cell receptor

Hayden. Nature 2012

T cell receptor

recognises

tumour cell

PD-L1 binds

PD-1, inhibiting

T cell response

A separate

therapy uses

antibodies that

bind PD-L1 on the

tumour cell

Antibodies

block inhibitory

signal to PD-1

T cellTumour

cellT cell

Tumour

cell

PD-L1PD-1

TCR MHC

PD-L1

PD-1

PD-L1

PD-1

Truth is: many cell types can be PD-L1+

IFN-g=interferon gamma

Chen, et al. Clin Cancer Res 2012

PD-L1

PD-L1

B7.1

B7.1

MHC I

B7.1

PD-1

PD-L1

CD28

TCR

PD-L1

PD-1

MHC I

TCR

PD-L1 PD-1

PD-L1

PD-1

PD-1PD-L2

Tumour

cell

CD8+ cytotoxic

T lymphocyte (CTL)

Tumour-associated

fibroblast

M2 Macrophage

Dendritic

cell

Th2

T cell

Treg

cell

Priming and

activation of T cells

PD-L1/PD-1-mediated

inhibition of tumour

cell killing

IFN-g-mediated

upregulation of

tumour PD-L1

Immune cell

modulation of T cells

PD-L2-mediated

inhibition of TH2 T cells

Stromal PD-L1

modulation of T cells

IFN-g

IFN-gR

TGF-b

IL-4/IL-13

T cell polarisation

Other NFkB PI3K

Shp-2

Shp-2

CARs and PD-1 at tumor–T cell immune synapses.

Morales-Kastresana A et al. Clin Cancer Res 2013;19:5546-5548


MHC

PD-L1

PD-1

PD-1

T-cellreceptor

PD-L2

T cell

NFκB

Other

PI3K

Tumor cell

IFNγ

IFNγR

Shp-2

Nivolumab

Nivolumab and Immune

Checkpoint Inhibition

• Nivolumab is a fully human IgG4 anti-PD-1 monoclonal antibody that selectively

blocks the interaction between PD-1 and PD-L1/PD-L2,1 restoring T-cell immune

activity directed against the tumor cell

1. Topalian SL, et al. N Engl J Med. 2012;366:2443-2454

25

And across tumour types, mutation load does not

correlate with clinical activity40%

20%

0%

28%2

0%2

6%1

25%3

0%1

20%5

31%4

0%1

~26%7

~21%8

33%9

29%6

ORR

Alexandrov, et al. Nature 2013

1. Brahmer, et al. N Engl J Med 2012; 2. Lesokhin, et al. ASH 2014; 3. Motzer, et al. N Engl J Med 2015; 4. Muro, et al. ESMO 2014

5. Chow, et al. ESMO 2014; 6. Ott, et al. WCLC 2015; 7. Powles, et al. Nature 2014; 8. Garon, et al. ESMO 2014; 9. Robert, et al. ESMO 2014

27

HCC and Programmed Death-1

(PD-1)

• HCC is typically an inflammation-associated cancer and can be immunogenic1

• Association of hepatitis C and hepatitis B infection with upregulation of PD-12,3

• Upregulation of PD-1 and the PD-1 immune checkpoint ligand, PD-L1, in HCC is

associated with poor outcomes4

• Blockade of PD-1 with monoclonal antibodies combined with immunostimulatory

monoclonal antibodies extended survival in a mouse model of HCC5,6

• Immune checkpoint inhibition (anti-CTLA-4) has shown encouraging activity in an early

clinical trial in HCC7

• PD-1 blockade with nivolumab may boost host immunity against HCC and improve clinical

outcomes

1. Hato T, et al. Hepatol. 2014;60:1776-1782

2. Xu P, et al. Gut Liver. 2014;8:186-195

3. Barathan M, et al. Apoptosis. 2015;20:466-480

4. Zeng Z, et al. PLoS One. 2011;6:e23621

5. Chen Y, et al. Hepatology. 2015;61:1591-1602

6. Morales-Kastresana A, et al. Clin Cancer Res.

2013;19:6151-6162

7. Sangro B, et al. J Hepatol. 2013;59:81-88

Phase 1/2 Safety and Antitumor Activity

of Nivolumab in Patients With Advanced

Hepatocellular Carcinoma (HCC): CA209-

040

Anthony B. El-Khoueiry,1 Ignacio Melero,2 Todd S. Crocenzi,3

Theodore H. Welling III,4 Thomas Yau,5 Winnie Yeo,5 Akhil Chopra,6

Joseph F. Grosso,7 Lixin Lang,7 Jeffrey Anderson,7 Christine de la Cruz,7 Bruno

Sangro2

1University of Southern California Norris Comprehensive Cancer Center, Los Angeles,

CA, USA; 2Clinica Universidad de Navarra and CIBERehd, Pamplona, Spain; 3Providence Cancer Center, Portland, OR, USA; 4University of Michigan, Ann Arbor,

MI, USA; 5University of Hong Kong, China; 6Johns Hopkins Singapore International

Medical Centre, Singapore; 7Bristol-Myers Squibb, Princeton, NJ, USA

Abstract LBA 101

29

Study Design

• Patients received nivolumab Q2W for up to 2 years (maximum of 48 doses), depending on response

– Imaging for disease assessment performed every 6 weeks

• A 3+3 design was used in the phase 1 dose escalation phase

• Here, we report interim results from the ongoing dose escalation phase and part of the expansion phase

Dose Escalation Expansion

Uninfected sorafenibprogressors

3 mg/kg (n=50)

HCV-infected

Expand at 3 mg/kg

(n=50)

HBV-infected

Dose TBD

(n=50)

Uninfected 0.3 mg/kg 1 mg/kg 3 mg/kg 10 mg/kg0.1 mg/kg

HCV-infected 0.3 mg/kg 1 mg/kg 3 mg/kg 10 mg/kg

Uninfected sorafenib-naïve or intolerant

3 mg/kg (n=50)

HBV-infected 0.3 mg/kg0.1 mg/kg 1 mg/kg 3 mg/kg 10 mg/kg

Baseline Patient CharacteristicsUninfected

(n=24)

HCV

(n=12)

HBV

(n=11)

Total

(N=47)

Age (years), median

(range) 63.5 (22–79) 67 (55–83) 62 (41–68) 64 (22–83)

Male, n (%) 18 (75) 7 (58) 9 (82) 34 (72)

Race, n (%)

White 20 (83) 10 (83) 1 (9) 31 (66)

Asian 2 (8) 2 (16) 10 (91) 14 (30)

Black 2 (8) 0 0 2 (4)

Extrahepatic

metastases, n (%) 17 (71) 7 (58) 9 (82) 33 (70)

Vascular invasion, n (%) 3 (13) 2 (17) 1 (9) 6 (13)

Child-Pugh Score, n (%)

5 21 (88) 9 (75) 11 (100) 41 (87)

6 3 (13) 2 (17) 0 5 (11)

7 0 1 (8) 0 1 (2)

AFP >200 μg/L, n (%)* 8 (33) 2 (17) 7 (64) 17 (36)

*Baseline AFP values were missing for 5 patients 30

Prior Treatment History

Treatment Type, n (%) Uninfecte

d (n=24)

HCV

(n=12)

HBV

(n=11)

Total

(N=47)

Prior surgical resection 14 (58) 8 (67) 9 (82) 31 (66)

Prior radiotherapy 6 (25) 1 (8) 1 (9) 8 (17)

Local treatment for HCC 12 (50) 8 (67) 9 (82) 29 (62)

TACE, TAE, or

radioembolization 11 (46) 8 (67) 7 (64) 26 (55)

RFA or PEI 5 (21) 4 (33) 4 (36) 13 (28)

Other 1 (4) 0 1 (9) 2 (4)

Prior systemic therapy 17 (71) 7 (58) 11 (100) 35 (75)

Prior sorafenib 15 (63) 6 (50) 11 (100) 32 (68)

TACE, transcatheter arterial chemoembolization; TAE, transcatheter arterial embolization; RFA, radiofrequency ablation;

PEI, percutaneous ethanol injection

31

Uninfecte

d

(n=21)

HCV

(n=11)

HBV

(n=10)

Total

Evaluable

*

(n=42)

Objective response, n (%) 3 (14) 4 (36) 1 (10) 8 (19)

Complete response 2 (10) 0 0 2 (5)

Partial response 1 (5) 4 (36) 1 (10) 6 (14)

Stable disease 10 (48) 5 (45)† 5 (50) 20 (48)

Progressive disease 8 (38) 2 (18) 4 (40) 14 (33)

Ongoing response, n (%) 3/3 (100) 3/4 (75) 0 6/8 (75)

Responses assessed by RECIST 1.1

*5 patients not evaluable: first disease assessment not yet performed in 4 patients, 1 patient died from clinical

progression before disease assessment†Patient with resolved HCV infection

Investigator-Assessed Best Overall

Response

32

Maximal Change in Target Lesions

From Baseline120

100

80

60

40

20

0

-20

-40

-60

-80

-100

Patients (N = 40)†

Ch

an

ge

in

Ta

rge

t L

es

ion

Fro

m B

as

eli

ne

, %

Uninfected

HCV

HBV

Confirmed response

*

* *

* ** *

* *

†2 uninfected patients not shown: 1 had disease progression before the first assessment; 1 had a maximal change of +23%‡Patient with resolved HCV infection

‡

33

Response Kinetics

Time Since First Dose, Months

Ch

an

ge

in

Ta

rge

t L

es

ion

Fro

m B

as

eli

ne

, %

180

0 3 6 9 12 15 18 21

-100

-80

-60

-40

-20

0

20

40

60

80

100 Months,

range

Uninfected

(n=21)

HCV

(n=11)

HBV

(n=10)

Total

Evaluable

(n=42)

DOR7.2* –

12.5*1.4* – 8.3* 11.9 1.4* – 12.5*

Duration of SD1.1* –

17.3*2.9† – 14.0 2.7* – 6.9* 1.1* – 17.3*

*Censored†Patient with resolved HCV infection

First occurrence of new lesion

34

+

Partial Response to Nivolumab

• 63 year-old male, uninfected HCC, Child-Pugh score A5

• No prior sorafenib or other treatment for HCC

35

BaselineAFP: 21,000 IU/mL

Week 6AFP: 283 IU/mL

Remove inhibitions

Release the brakes Press the gas pedal to the metal

Enforce costimulation

• CD137, also known as 4-1BB, is a surface glycoprotein involved in T cell costimulation

• Its cognate ligand is CD137L, which is expressed on APCs

• Functions: T cell proliferation, inhibition of apoptosis, enhances cytotoxic activity, cytokine production

• Therapeutic target: treatment with agonist anti-CD137 monoclonal antibodies can overcome tumour antigen tolerance

• Anti-human CD137 agonist monoclonal antibodies are undergoing phase I/II clinical trials

CD137 (4-1BB, TNFRSF9)

T cellAPC

4-1BBL 4-1BB

Melero, et al. Trends Pharmacol Sci 2008

• Resistance to apoptosis• Proliferation• Gain effector functions• Differentiation to memory cells

Early cytokine production

Early tumour cell killing

Provision of tumour cell debris for cross-priming

Tumour antigencross-presentation

Proinflammatory moleculeslymphocyte infiltration?

Inhibition/activationin vivo paradox

Antigen-independentactivation

Controversial effects

on regulatory function

CytokinesecretionIL-13 and

IFN-g

Anti-CD137mAb

Memory

CD8+ T cell

Dendritic

cell

Activated

NK cell

Activated

CD4+ T cell

T regs?

IKDC?

NKT

cell

Tumour endothelium

Activated

CTL

Melero, et al. Trends Pharmacol Sci.2008

NF-kBJNK, AP-1

p38 MAPK

-Transcriptional control

Anti-CD137

CD137 (4-1BB)

TRAF-2

Ubiquitin

TAK1

4-1BB signalsome?

TRAF-1

Figure 2

Early signal transduction events from CD137.



SYNERGY

syn-ergos, συνεργός, meaning 'working

together'.

http://en.wiktionary.org/wiki/%CF%83%CF%85%CE%BD%CE%B5%CF%81%CE%B3%CF%8C%CF%82

Strategies for immunotherapy combinations.



-OX40

Radiotherapy

Chemotherapy

Vaccination

T-reg depletion/

inactivation

Adoptive T-cell

immunotherapy

–Clinical Standard

–Clinical Trials

–Preclinical Studies

Antiangiogenic

therapy

-TIM-3

-LAG3

-CTLA-4

-CD137

-PD1

-PD-L1

-CD40

Virotherapy

or intratumoral

PAMPS

Spontaneous HCC (c-mycOVA75/tTALAP)

Ney et al, Hepatology 2009

HC

CLitte

rmate

Week 5

H&E OVA

Anti-CD137 + anti-PD-1 + anti-OX40

12/39

Combo3

d -21 d 0 d21

breeding Birth weaning

Dox

d25

TREATMENT

mAbs mAbs

Survival

Rat IgG

Combo3

ACT

ACT + Combo3

Rat IgG ACT + Combo3Combo3

42,42 mm2 11,18 mm2 No tumor

4/11

2/10

Week 6

TILs

CD8+

alive/total

Combo3 efficacy is dependent on CD8+ T cells

GzmB Perforin FasL

Rat IgG

Combo3

17,8% 12,7

462 185

15% 4,5

143 23

1,4% 0,1

12 1

69,4% 6

2083 225

63,5% 0,4

371 7

3,1% 0,6

39 7

Effector machinery is enhanced upon Combo3 Tx

Gated on CD8 TILs

Synergy with adoptive T cell therapy

0 1 0 2 0 3 0 4 0 5 0 6 0 7 0 8 0

0

1 0 0

2 0 0

3 0 0

4 0 0

B 1 6 -O V A w /o T x

O T 1 + R a t Ig G

O T 1 w /o a b

0 /50 /50 /5

T im e (D a y s )

Tu

mo

r a

re

a (

mm

2)

0 1 0 2 0 3 0 4 0 5 0 6 0 7 0 8 0

0

1 0 0

2 0 0

3 0 0

4 0 0

0 /6

T im e (D a y s )

0 1 0 2 0 3 0 4 0 5 0 6 0 7 0 8 0

0

1 0 0

2 0 0

3 0 0

4 0 0

1 1 /1 1

T im e (D a y s )

0 1 0 2 0 3 0 4 0 5 0 6 0 7 0 8 0

0

1 0 0

2 0 0

3 0 0

4 0 0

0 /5

T im e (D a y s )

0 1 0 2 0 3 0 4 0 5 0 6 0 7 0 8 0

0

1 0 0

2 0 0

3 0 0

4 0 0

5 /6

T im e (D a y s )

Tu

mo

r a

re

a (

mm

2)

0 1 0 2 0 3 0 4 0 5 0 6 0 7 0 8 0

0

1 0 0

2 0 0

3 0 0

4 0 0

0 /6

T im e (D a y s )

0 1 0 2 0 3 0 4 0 5 0 6 0 7 0 8 0

0

1 0 0

2 0 0

3 0 0

4 0 0

0 /1 1

T im e (D a y s )

0 1 0 2 0 3 0 4 0 5 0 6 0 7 0 8 0

0

1 0 0

2 0 0

3 0 0

4 0 0

1 /7

T im e (D a y s )

C o n tr o lsN o n -T C R tr a n s g e n ic

a c t iv a t e d T c e l ls + C D 1 3 7

O T -1 + C D 1 3 7

(T r e a te d o n d a y + 3 )

O T -1 + C D 1 3 7

(T r e a te d o n d a y + 7 )

O T -1 + C D 1 3 7

(C D 4 d e p le t io n )

O T -1 (C D 1 3 7 K O )

+ C D 1 3 7

O T -1 + C D 1 3 7

in C D 1 3 7 K O

O T -1 + C D 1 3 7

in R A G 1 K O

A B C D

E F G H

Figure 2.

0

5 0 0

1 0 0 0

1 5 0 0

2 0 0 0

M o u s e :

O T -1 :

A b :

W T C D 1 3 7 K O

1 D 8 R a t Ig G

2 3 7 ,5

0 ,0 0 69 ,5 0 5 4 2 7

p = 0 ,0 0 6 3

W T

W T W T W T

1 D 8

C D 1 3 7 K O

W T

1 D 8

MF

I

0

3 5 0

7 0 0

1 0 5 0

1 4 0 0

1 9 9 ,5

p = 0 ,0 0 2

p = 0 ,0 1 0 9

1 4 4

M o u s e :

O T -1 :

A b :

W T C D 1 3 7 K O

1 D 8 R a t Ig G

W T

W T W T W T

1 D 8

C D 1 3 7 K O

W T

1 D 8

0 ,0

1 6 ,5

MF

I

0

2 0 0

4 0 0

6 0 0

8 0 0

2 1 2

2 8 ,5 0 ,0

1 4 8

p = 0 ,0 2 6 7

M o u s e :

O T -1 :

A b :

W T C D 1 3 7 K O

1 D 8 R a t Ig G

W T

W T W T W T

1 D 8

C D 1 3 7 K O

W T

1 D 8

MF

I

0

1 6 0 0

3 2 0 0

4 8 0 0

6 4 0 0

1 3 7 2

p = 0 ,0 0 1 8

p < 0 ,0 0 0 1

p = 0 ,0 0 0 3

p = 0 ,0 3 1 1

1 8 2 4

2 7 4 ,5

M o u s e :

O T -1 :

A b :

W T C D 1 3 7 K O

1 D 8 R a t Ig G

W T

W T W T W T

1 D 8

C D 1 3 7 K O

W T

1 D 8M

FI

0

6 5 0 0

1 3 0 0 0

1 9 5 0 0

2 6 0 0 0 n s

5 2 32 5 2 0 ,5 1 7 1

M o u s e :

O T -1 :

A b :

W T C D 1 3 7 K O

1 D 8 R a t Ig G

W T

W T W T W T

1 D 8

C D 1 3 7 K O

W T

1 D 8

MF

I

0

3 0 0 0

6 0 0 0

9 0 0 0

1 2 0 0 0n s

2 0 0 ,00 ,0 1 5

M o u s e :

O T -1 :

A b :

W T C D 1 3 7 K O

1 D 8 R a t Ig G

W T

W T W T W T

1 D 8

C D 1 3 7 K O

W T

1 D 8

MF

I

S u r fa c e C D 1 0 7 a

S u r fa c e K L R G 1

In tra c e llu la r IF N g

T ra n s fe r re d C D 8+

E n d o g e n o u s C D 8+

A

B

C

Figure 3.

MS

D(µ

m2)

A B C

D

Unconfined tracks, low dwell time per tumour cell

α-CD137

Confined tracks, concentrated Dwell timehigh dwell time per tumour cell

Ctrl

0h 1h

0

50

100

150

200

CT

L :

tu

mo

ur

ce

llco

nta

ctd

ura

tio

n(m

in)

0

Tumour: F10 F10/OVA

2

4

8

6

10

12

Sp

ee

d(µ

m/m

in)

OT1OT1 + αCD137

OT1OT1 + αCD137

OT1OT1 + αCD137

0 20 40 60 80

dt (min)

0

1000

2000

3000

Weigelin, et al. PNAS in press

Tumor

Proliferation

Blood

vessel

Activation of

endothelial cells

Enhanced CTL effector

functions

Costimulation

Killing

A B

C

00:00 h 00:40 h 01:06 h

CTL mitosis00:00 h 00:14 h 00:32 h

Tumor cell apoptosis

OT1 CD8 T cells Blood vessels

B16F10/OVA melanoma cell nuclei

CD8 T cells B16F10/OVA melanoma cells

Autofluorescent tissue

Hot spots for extravasationOverview tumor lesion

500 µm

CTL migration tracks

Enrichment of potent

effector cells

Costimulation during

activation

Chimeric receptors

containing CD137cyt

Ligand binding

Transmembrane

Costimulatory

Signaling

CD137 -derived

Dendritic cell Migration track

CD8 T cellTumor cellα-CD137 mAb

TILAPC

Proliferation

Effector function

Cell culture flaskDomains:

Imm

uno

-magnetic

cell

sort

ing

Undesired cells

TIL

CD137 and adoptive T cell transfer. A perfect marriage?

PROFESSIONALCROSSPRIMING DENDRITIC CELLS

0 2 0 4 0 6 0

0

1 0 0

2 0 0

3 0 0

4 0 0

0 / 2 8

0 2 0 4 0 6 0

4 / 2 3

0 2 0 4 0 6 0

1 6 /2 3

0 2 0 4 0 6 0

8 / 9

0 2 0 4 0 6 0

0 / 1 6

0 2 0 4 0 6 0

0

1 0 0

2 0 0

3 0 0

4 0 0

0 / 2 4

0 2 0 4 0 6 0

0 / 1 6

0 2 0 4 0 6 0

0 / 9

Untreated αCD137 αPD-1 αCD137 + αPD-1

WT

Batf3-/-

Days post-inoculation

Tu

mo

r si

ze (

mm

2)

0 2 0 4 0 6 0 8 0

0

2 0

4 0

6 0

8 0

1 0 0 8/9

16/23

4 /23

W T C D 1 3 7 + P D - 1

W T C D 1 3 7

W T P D - 1

W T c o n tro l

***

0 2 0 4 0 6 0 8 0

0

2 0

4 0

6 0

8 0

1 0 0

B a t f3- / -

C D 1 3 7 + P D - 1

B a t f3- / -

C D 1 3 7

B a t f3- / -

P D - 1

B a t f3- / -

c o n t r o l

**

WT Batf3-/-

Per

cent su

rviv

al

Days post-inoculation Days post-inoculation

Crosspriming DC absolutely needed!

0 10 20 300

50

100

150

200

Control mAb

13%

CD137

Avera

ge t

um

or

siz

e (

mm

2)

0 10 20 300

50

100

150

200

250

Batf3-/-


Avera

ge t

um

or

siz

e (

mm

2)

0 10 20 300

50

100

150

200

Batf3-/-


Avera

ge t

um

or

siz

e (

mm

2)

0 10 20 300

50

100

150

200

33%

FLT3L+PolyIC FLT3L+PolyIC + mAb

-PD-1

Avera

ge t

um

or

siz

e (

mm

2)

IMMUNOAVATAR

MICE PROJECTS

0

50

100

150

hPBMCs

hIgG4hCD137

hPD-1

4 7 21Time (Days) 4 7 21 4 7 21 4 7 21

+ + ++ + + + + ++ + +- - -- + +- + +

- - -- - -- + +

- - -- + +- - -

- - -- - -

- + +

h-I

FNg

(pg

/mL

)

Xenografted syngenic gastric cancer with autologous PBMCs experiments

Sacrifice(day 50)

5·105

PBMCs(i.p.)

7mmx7mmTUMOR

TRANSPLANT

0 5 6 12 19 26 33 40 48

200g Urelumab 200g Nivolumab (iv)TUMOR:

-FACS ANALYSES

-IHQ

Immunostimulatory monoclonal antibodies anti-hCD137 (urelumab) and anti-hPD-1 (nivolumab) alone or in combination show antitumoractivity against a xenografted human colon cancer mediated by transferred allogenic human PBMC.

Xenografted syngenic gastric cancer with autologous PBMCs experiments

Co

mb

oα

-hC

D1

37

α-h

PD

-1C

on

tro

l

CD3/CD8/CD20/CK/DAPI




IgG4

-hCD13

7

-h

PD-1

Com

bo0.00

0.01

0.02

0.03

0.04

Tumor compartment

*

rati

o h

CD

3/t

ota

l p

are

cn

ch

ima c

ells

IgG4

-hCD13

7

-h

PD-1

Com

bo0.0

0.2

0.4

0.6

0.8Stromal compartment

rati

o h

CD

3/ to

tal str

om

a c

ells

Multiplexed immunofluorescence microphotographs showing the architecture of tumor xenografts

MINECO

Gob Navarra

EU FP7 & H2020

AECC

AICR

Fundación BBVA

Fundación CAN

OUR GRATITUDE GOES TO:

Patients and

their families

Central facilities at

CIMA and CUN

combination opportunities in immunotherapy with ... · hospital universitario reina sofía,...

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