Immunity Agreement: Re-engineering the Immune Response to ... · Hingorani et al., J Clin Oncol...

6/3/2014

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Immunity Agreement: Re-engineering the Immune Response to Treat Pancreas Cancer

Sunil R. Hingorani, M.D., Ph.D. Fred Hutchinson Cancer Research Center University of Washington Medical Center

Seattle Cancer Care Alliance May 16, 2014

Disclosures

• I have no financial disclosures to report

• I will be presenting preliminary data on the investigational use of a stromal disrupting agent

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Cancer cells

Immune cells, fibroblasts, matrix

Pancreas cancer as a “solid tumor organ”: multi-faceted stromal response

CC

CD8

Mesenchyme

Shh TGFb

TGFb

IDO IL-10 PDL1

PDGF FGF

collagens

PSC

CXCL12

Olive, 2009, Science Kraman, 2010, Science Ene-Obong, 2013, Gastroenterology Feig, 2013, PNAS

GM-CSF G-CSF

CCL2

TAM Treg

MDSC

CD8 TGFb

IL-10 CTLA4

NO ARG1 ROS

NO ARG

PDL2

Hematopoietic

Clark, 2007, Cancer Research Beatty, 2011, Science Bayne, 2012, Cancer Cell Pylayeva-Gupta, 2012, Cancer Cell Porembka, 2012, Can Immunol Immunother Sanford, 2013 Clin Canc Res Stromnes, 2014, Gut

HA

fibrillary collagen

IFP

vessel

ECM

cytotoxic

HA

Provenzano, 2012, Cancer Cell Jacobetz, 2013, Gut

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In vitro

Crossing the translational divide: matching model systems to clinical strategies

Hingorani and Potter, Sci Transl Med 2013

In vitro


Heterotopic


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In vitro


Orthotopic


In vitro


PDX


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Pla

tfo

rm

De

scri

pti

on

P

roce

sse

s M

od

ele

d

Stre

ngt

hs

Lim

itat

ion

s

Heterotopic

Human cell lines in immune-deficient host (murine cells can be

studied in syngeneic immune-competent host)

Growth (+/- metastasis) of

invasive disease

Human tumor cells; relative

ease

Heterologous, cross-species

growth of clonally selected

lines; absent immunity

Orthotopic


invasive disease

Human tumor cells; growth in

orthologous organ

Cross-species growth of

clonally selected lines; absent

immunity

Autochthonous (preinvasive)

GEMM of spontaneous

PDA at preinvasive stage

Initiation and progression of

preinvasive disease

Spontaneous co-evolution of tumor epithelium and microenvironment in

native organ; recapitulation of clinical and physiological disease syndromes;

amenable to rigorous correlative studies

Murine cancer; multifocal disease initiation; labor-

and resource-intensive

Autochthonous (invasive/metastatic)

GEMM of spontaneous

PDA at invasive and metastatic stages

Progression of invasive and

metastatic disease with associated clinical sequelae

Murine cancer; multifocal disease initiation; labor-

and resource-intensive

PDX

Patient-derived tumor pieces in

immune deficient host


invasive disease

Human tumor cells with some

preserved human stromal

elements

Heterologous, cross-species

growth; absent immunity; loss of architecture and

mechanics


Pancreas cancers are pale and fibrous

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WT KPC

Lect

in/S

HG

Le

ctin

/Do

xo

Vasculature in PDA is sparse, collapsed and poorly functional (intravital imaging)

Provenzano et al., Cancer Cell 2012

MPLSM

Real-time monitoring of Interstitial Fluid Pressure (IFP)


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IFP is low in normal tissues but extremely high in autochthonous PDA

normal

KPC

WHY?


Characterizing stromal cells and matrix deposition in PDA


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Hyaluronic acid (HA), or hyaluronan

• Large, linear glycosaminoglycan composed of repeating units of NAG and glucuronic acid

• Has viscoelastic properties, and figures prominently in tissue architecture, integrity and malleability

• Imbibes and “traps” large amounts of water in both mobile and immobile fluid phases

Systemic administration of pegylated hyaluronidase degrades intratumoral HA and alters microenvironment

Baseline PEGHAse

HA

BP

H

&E

CD

31

24 hours

Provenzano et al., Cancer Cell 2012 Jacobetz et al., Gut 2013

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WT KPC KPC + PEGHAse Le

ctin

/SH

G

Lect

in/D

oxo

Enzymatic degradation of HA improves functional perfusion (intravital imaging)

MPLSM


0

50

100

150

200

250

300

Pre treatment Post 6 wks treatment

H-S

co

re

HA High

HA Low

HA-rich metastases can be depleted with systemic PEGPH20

Pre-treatment biopsy

h m

H+E HABP

Post-treatment biopsy

h

m

H+E HABP

Clinical Vignette #1

Hingorani et al., ECCO 2013 (abstr 2598) Hingorani et al., J Clin Oncol 2013 (abstr 4010)

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• 33 % reduction in size across three target lesions at EOC1 • A representative target lesion (TL3; red arrow) demonstrated a 47% reduction in

cross-sectional size and 78% reduction in volume • Representative non-target lesions (white arrows) also improved

Combination therapy can induce regression of primary tumors and metastases

TL3 D:19.3 mm V:1.5 cm3

TL3 D:10.2 mm V:0.33 cm3

EOC 1 (Sum of Target Lesions) = 56.1 mm Baseline (Sum of Target Lesions) = 83.2 mm

Baseline: Mean = 0.14 min–1 Median = 0.10 min–1

24-hr Post 1st Dose: Mean = 0.20 min–1 Median = 0.17 min–1

Vignette #2: Increased perfusion (Ktrans) after PEGPH20


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Combination therapy can decrease metabolic activity in primary tumors and metastases

Vignette #2: PMR at EOC1 (52% reduction in 18FDG-PET)

Bas

elin

e SU

Vm

ax

(Su

m o

f Ta

rget

Les

ion

s)

Axial Fused 18FDG-PET/CT

26

.3

12

.6

2o Endpoints: Objective Response

• PEGPH20/Gem ORR = 42% (21.9%-61.4%) (95% CI) (1.6 and 3.0 µg/kg dose levels)

• Total of 24 patients (20 patients with at least 1 baseline and 1 follow up scan)

• 6 patients had clinical progression in Cycle 1 (no follow up CT scans)

-70

-60

-50

-40

-30

-20

-10

0

10

20

1.0 g/kg

3.0 g/kg

1.6 g/kg

* * * * * * * * * *

* PR (n=10)

Ch

an

ge f

rom

Baselin

e in

Targ

et

Lesio

ns (

%)


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Current status of systemic enzymatic therapy (PEGPH20) for PDA

• Phase 1b Trial completed and analyzed • Two randomized, placebo-controlled Phase 2 trials have opened

nationwide to test the two most recent FDA-approved combination cytotoxic regimens (Abraxane+Gem and mFOLFIRINOX) with and without PEGPH20

• Plans to incorporate clinical science correlates in separate of

cohorts of patients

Hematopoietic Compartment

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Why does the immune system fail?

• Cancer cells are not different enough from normal cells

• Immune cells do respond at first and then are shut down to prevent “auto-immunity”

• Something else

Cancer immunosurveillance and immunoediting

Modified from Dunn, Old, and Schreiber, Immunity 2004

Elimination Equilibrium

CD4+ CD8+

NK

Escape

CD8+

CD4+

NK

Treg

CD4+

CD8+

NK

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Cancer immunosurveillance and immunoediting

Modified from Dunn, Old, and Schreiber, Immunity 2004

Elimination Equilibrium

CD4+ CD8+

NK

Escape

CD8+

CD4+

NK

Treg

CD4+

CD8+

NK

1. Early infiltration of TAMs and Treg followed by MDSC 2. Absence of any

effective immunity

TAM Treg MDSC

Clark et al. Cancer Res 67:9518-27, 2007

Stromal reaction in pancreas cancer: Immune Cells

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Stromnes et al., Gut 2014

1 0 7

# Tr

eg 1 0

6

1 0 5

1 0 4

1 0 3

n.s. *

nl Pre PDA

# M

acro

ph

ages

1 0 7

1 0 6

1 0 5

1 0 4

1 0 3

n.s. *

nl Pre PDA

# M

DSC

1 0 7

1 0 6

1 0 5

1 0 4

1 0 3

n.s. *

nl Pre PDA

# N

K

1 0 7

1 0 6

1 0 5

1 0 4

1 0 3

n.s. n.s.

nl Pre PDA

Distinct kinetics of tumor infiltration by immune cell populations

Treg Mac MDSC NK

Clark et al., Canc Res 2007


Normal Preinvasive PDA

Ly6

C

CK

D

AP

I Ly

6C

C

K

DA

PI

Ly6

G/

*

MDSC infiltrate in the transition between preinvasive and invasive disease

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CD8

(-)

88.6

CFSE

Sple

en

PD

A

44.8

54.5

Gr-MDSC inhibit CD8+ T cell proliferation


Gr-MDSC induce CD8+ T cell apoptosis

CFSE

An

nex

in-V

(-)

40.4 5.5

(+)

Gr-MDSC

Spleen PDA

18.9 33.8

(+) (+)

0

1 0

2 0

3 0

4 0

Gr-MDSC

% A

nn

exin

-V

Spl PDA (-)

**

*

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Basic Media PDA-CM

Basic Media PDA-CM

75.1 38.6

An

nex

in-V

Gr-1

Tumor epithelial cells promote Gr-MDSC survival

Pylayeva-Gupta et al., Cancer Cell 2012 Bayne et al., Cancer Cell 2012 Stromnes et al., Gut 2014


0

2

4

6

8

1 0

2 0

4 0

6 0

8 0

1 0 0 PDA Metastasis

Fold

ch

ange

/ p

rein

vasi

ve

Tumor epithelial cells secrete critical chemokines and cytokines

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Blood

KPC Normal KPC + 1A8

53.4 0.2 0.01

CD11b

Gr-

1

Targeted antibody-mediated depletion of endogenous Gr-MDSC

Proposal: Removing the Cloaking Device in Pancreas Cancer will Awaken the Immune Response

CD8+

CD8+

CD8+

CD8+

CD8+

CD8+ CD8+

CD8+

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CD

8

CK

D

AP

I Control 1A8

*

Targeted depletion of endogenous Gr-MDSC results in increased intratumoral CD8+ T cells


Targeted depletion of endogenous Gr-MDSC induces specific increase in intratumoral CD8+ T cells

0

2

4

6

8

10

% C

D8

T c

ells

n.s. *

Spleen PDA

n.s. *

10 5

10 6

10 7

# C

D8

T c

ells

Spleen PDA

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0

10

20

40

% K

i67

+ C

D8

cel

ls

* ***

Spleen PDA

30

% C

D6

9+

CD

8 c

ells

* *

Spleen PDA 0

10

20

50

30

40

Infiltrating intratumoral CD8+ T cells show signs of activation


Control 1A8

CC

3

Ki6

7

*

Control 1A8

# C

C3

+/4

0X

fie

ld

0

5

10

15

20

n.s.

Control 1A8

# K

i67

+/4

0X

fie

ld

0

10

20

30

40

Targeted depletion of Gr-MDSC results in increased tumor cell apoptosis

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Increased intratumoral CD8+ T cell accumulation is accompanied by stromal remodeling

Control 1A8

H&

E M

asso

n’s

M

ova

t’s

CD

31

CK

DA

PI

Conclusions

• Multiple stromal processes contribute significantly to progression and resistance of PDA

• Systemic administration of a catalytic agent with a prolonged circulatory half-life can remodel the TME, altering stromal architecture and mechanics and enabling delivery of therapeutics

• Targeted depletion an MDSC subset (Gr-MDSC) can unmask PDA to endogenous adaptive immune response

• Diverse strategies for “stromal disruption” represent novel, and perhaps essential, approaches to eradicating PDA

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Acknowledgments

Hingorani Lab Scott Brockenbrough Markus Carlson Carlos Cuevas Ashley Dotson Kathy DelGiorno Libing Feng Nathan Lee Justin Mirus Joe Ryan Ingunn Stromnes Shelley Thorsen Marty Whittle Peter Williams

Hingorani Lab - Past Paolo Provenzano Kamel Izeradjene Geetha Rani Amy Chang Philamer Calses Hsin-Pin Lin Melissa Best Catherine Gard Natalie Wong Randi Simmons

Giles and Elise Mead Foundation Lustgarten Foundation Jeffrey Rosenzweig Foundation Safeway Fund Tagney-Jones Funds

Halozyme Therapeutics Gregory Frost Michael Shepard Joy Zhu Curtis Thompson

FHCRC Phil Greenberg

http://www.nci.nih.gov/

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