Wong raymond symposium2016_present

Harnessing the Full Potentialof Immunotherapy by Targeting the Tumor Microenvironment

Raymond Wong, Ph.D.Principal Investigator

US FDA approved immune checkpoint blockers: 2011 ipilimumab (anti-CTLA-4) melanoma 2014 pembrolizumab (anti-PD-1) melanoma 2014 nivolumab (anti-PD-1) melanoma 2015 pembrolizumab (anti-PD-1) lung cancer 2015 nivolumab (anti-PD-1) lung cancer 2015 nivolumab + ipilimumab combo melanoma 2016 nivolumab (anti-PD-1) kidney cancer 2016 nivolumab (anti-PD-1) Hodgkin’s lymphoma 2016 atezolizumab (anti-PD-L1) bladder cancer

PD-1 or PD-L1 clinical trials: ≥ 700 registered trials for PD-1 or PD-L1 blockers. ≥ 10 PD-1 or PD-L1 trials for mesothelioma.

Recent Breakthroughs inCancer Immunotherapy

Programmed death-1 (PD-1) and programmed death ligand-1 immune checkpoints.

PD-1/PD-L1 Immune Checkpoint Blockers for Cancer

PD-1 or PD-L1blockers

TCR

MHC

PD-L1

CD80PD-L1

PD-L1

PD-1

PD-1CD80

PD-L1

T Cell

Antigen Presenting Cell(lymphoid organs)

Tumor “Microenvironment”(periphery)

Nivolumab / pembrolizumab / atezolizumab. Extended overall survival. Tumor shrinkage in select cases (15-25%).

• For most cancers complete remission is rare. • Nivolumab + ipilimumab = 10-20% complete remission in melanoma (Larkin, et

al., 2015 & Postow et al., 2015). Responding patients experience durable benefit that can last years after

stopping treatment.

Pembrolizumab (PD-1) in pleural mesothelioma (AACR 2015): Partial response in 28% patients (25 patients treated; Alley, et. al).

Avelumab (PD-L1) in pleural/peritoneal mesothelioma (ASCO 2016): Partial response in 9.4% patients (53 patients treated; Hassan, et. al).

Clinical Effects of PD-1/PD-L1 Blockers

Galectin-9

TCR

MHCPD-L1

PD-1/PD-L1blockers

PD-L1upregulate

CD80

T cell

Tumor

Adaptive Immune Resistance inthe Tumor Microenvironment

PD-1 TIM-3

upregulate

Modified from Wong et al., 2016

Combination Immunotherapyto Combat Immune Resistance

PD-1 blocker“common denominator”

CTLA-4 blocker Other Biological Response Modifiers

TIM-3 blocker PD-L1 blocker

Biological Response Modifiersfor Enhancing T Cells

Modulate the activity of T cells in tumor tissue: Promote T cell trafficking (influx to tumors)

• Chemokines Promote T cell survival and effector function

• Cytokines

Tumor-targeted delivery methods: Novel fusion proteins Nanoparticles Mesenchymal stem cells

• Natural tumor homing properties• Possibly “universally compatible”

Multi-potent can develop into different cell types Fat Cartilage Bone Muscle (e.g., heart)? Neurons? Blood vessels?

Can be harvested from: Bone marrow Body fat Umbilical cord blood Term placenta

Mesenchymal Stem CellProperties

Tissue maintenance / regeneration

Mesenchymal stem cells in culture

Pre-Clinical Efficacy ofMSCs in Mesothelioma Model

Sage et al., 2014

Intrapleuraltreatment

Systemic treatment

Isolating Mesenchymal StemCells From Primary Tissue

Digest / Separate

Adherent Cell Mixture+ MSC Growth Medium

~4 weeks(clear unwanted

cells)Expanded / Pure MSCs

Cryopreserve

Placenta / Bone Marrow (or other sources)

Engineered MSCs toProduce Human Cytokines

Producing MSC prototype therapeutics: Adenoviral-mediated genetic engineering of mesenchymal stem cells.

• Interleukin-12• Interferon-α

Adenovirus-IFN-αor

Adenovirus-IL-12

MSCs

IFN-α or IL-12 secretion

In Vitro Screen

Enhance human T cells?

IL-12-Producing MSCsBoost Human T Cells

Ad-IL-12 Ad-IFN-αAd-Mock(control)

Unmodified(control)

CD8

CFSE proliferation

77.4% 22.8% 34.1% 39.2%

CD4

19.4% 10.5% 12.1%4.71%

Ad-IL-12 Ad-IFN-αAd-Mock(control)

Unmodified(control)

0.1

1

10

100

1000

10000 Mock Virus

Ad-IFN-a2IL-12

Alterations in MSCSecretome Biology

FGF-

2Eo

taxi

nTG

F-a

G-CS

FFl

t-3L

GM-C

SFFr

acta

lkin

eIF

N-a

2IF

N-g

GRO

IL-10

MCP

-3IL-

12(p

40)

MDC

IL-12

(p70

)IL-

13IL-

15sC

D40L

IL-17

AIL-

1RA

IL-1a

IL-9

IL-1b

IL-2

IL-3

IL-4

IL-5

IL-6

IL-7

IL-8

IP-1

0M

CP-1

MIP

-1a

MIP

-1b

TNF-

aTN

F-b

VEGF

EGF

Ad-IL-12

IFN-α

Fold

Cha

nge

vs. C

ontr

ol

Pleural Effusion After P/D Surgery in Meso Patients

0

2000

4000

6000

8000

10000

12000

14000

16000

18000

20000 Meso Pt #1

FGF-

2

TGF-

aG-

CSF

Flt-3

LGM

-CSF

Frac

talk

ine

IFN

-a2

IFN

-gGR

OIL-

10M

CP-3

IL-12

(p40

)M

DCIL-

12(p

70)

IL-13

IL-15

sCD4

0LIL-

17A

IL-1R

AIL-

1aIL-

9IL-

1bIL-

2IL-

3IL-

4IL-

5IL-

6IL-

7IL-

8IP

-10

MCP

-1M

IP-1

aM

IP-1

bTN

F-a

TNF-

bVE

GF

EGF

Eota

xin

Meso Pt #2Meso Pt #3Control #1Control #2

pg /

ml

No detectable IL-12

Other Biological Response Modifiers

Immune checkpointblockersT cell activity

- LIGHT- CCL chemokine family - CXCL chemokine family

- IL-12- IL-2- IL-15

- Soluble CD80- High affinity PD-1 variant

T cell trafficking

Suicide gene

Clinical safety testing

+

Key Points Immunotherapy rapidly moving forward as new option for treating

diverse cancers. PD-1 / PD-L1 immune checkpoint blockers. Combination immunotherapies moving forward.

Many cancer patients do not respond to immune checkpoint blockers (≥ 50%). Complete regression / disease remission still rare. Countering adaptive immune resistance via combination immunotherapy is

likely key to improve tumor responses.

Mesenchymal stem cells as vectors for countering immune resistance in tumor microenvironment. Deliver immunotherapeutic agents specifically to tumor tissue.

Acknowledgements

Pacific Mesothelioma CenterIrina Ianculescu, PhDSteven ChingTonya LeeBlair Kimble

UCLA / West Los Angeles VARobert Cameron, MD

Research FundingPacific Heart, Lung & Blood InstituteRichard M. Shulze Family FoundationH.N. & Frances C. Berger FoundationKazan McClain Partners’ Foundation

Personalized vs. Off-The-ShelfStem Cell Manufacturing

Personalized Therapy

10 weeks

Off-The-Shelf Therapy

Healthy DonorPatient

ImmediatelyAvailable

Compatibility?