IS VISTA AN ACTIONABLE IMMUNE

CHECKPOINT TARGET IN KIDNEY CANCER?

Kathleen Mahoney, M.D., Ph.D.Instructor of Medicine, Harvard Medical School

Attending, Beth Israel Deaconess Medical Center

Dana Farber Cancer Institute

September 13, 2019

• No relevant financial relationships

• There will be no discussion about the use of products

for non-FDA approved indications in this presentation

Disclosures

Adapted from Mahoney et al. Kidney Cancer, 2nd ed. 2015

Tumor cell

PD-L1

PD-1

PD-L1MHCI

MHCII

B7-1

LAG3PD-L1

B7-1

PD-1

CTLA-4CD-8

CD28

T cell

Antigen

Presenting

cell

Ipilimumab – initial FDA approval 2011 Melanoma

Pembrolizumab – initial FDA approval 2014

Nivolumab – initial FDA approval 2014

Melanoma, Non-small cell lung (NSCLC), H&N…

Melanoma, Kidney, Hodgkin lymphoma, H&N, Bladder…

Atezolizumab – initial FDA approval 2016

Durvalumab – initial FDA approval 2017

Avelumab – initially FDA approval 2017

Cemiplimab – initial FDA approval 2018

Sintilimab (IBI308) – China 2018

Bladder cancer, NSCLC

Bladder cancer, NSCLC

Merkle cell cancer, Bladder cancer

Head & Neck cancers, CSCC

Hodgkin lymphoma

PD-1/PD-L1 blockade has FDA approval for 14 tumor types.

Ipilimumab and Nivolumab – first combination immunotherapy Melanoma and Kidney Cancer

(intermediate and high risk RCC)

Immune checkpoints regulate T cell function.

Adapted from Mahoney et al. NRDD 2015

ipilimumab

nivolumab

Tumors cells may express immune checkpoints.

ipilimumab

nivolumab

Adapted from Mahoney et al. NRDD 2015

There are higher levels of many B7 family proteins,

such as VISTA, in kidney cancer than normal kidney tissue

Mahoney, Freeman

PD-L1 PD-L2 B7H3 VISTA HHLA2B7H4 BTNL2 ICOSL CD8 CD14

VIS

TA

PD

-1

PD

-L1

PD

-L2

Mostly

specimens

from

patients

with clear

cell RCC

MIXED

papillary and

ccRCC

Mostly

papillary

RCC

Mostly

Chromophobe

VISTA is highly expressed across human kidney cancer.

Mahoney, Obi, Culhane

VISTA can be expressed on T cells.

Tumor

cellMHCI

MHCIIB7-1

VISTACD-8CD28

T cell

Antigen

Presenting

cell

VISTA

MDSC

Tumor associated

macrophage

VISTA

VISTA

Kakavand et al. Mod Path 2017

Xie, et al. Cancer Imm Imm 2018

Cadena, et al Front Imm 2019

VISTA-fusion proteins inhibits T cells function,

indicating an inhibitory receptor in T cells.

Tumor

cell

R-X

MHCI

MHCIIB7-1

VISTA

R-X

CD-8CD28

T cell

Antigen

Presenting

cell

R-X

MDSC

Tumor associated

macrophage

R-X

VISTAVISTA

VISTA

Wang et al. (Noelle) J Exp Med 2011

Lines et al. (Noelle) Cancer Res 2014

Prodeus et al JCI Insight 2017

However, VISTA is highly expressed on myeloid cells.

Tumor

cell

R-X

VISTAMHCI

MHCIIB7-1

VISTA

R-X

CD-8CD28

T cell

Antigen

Presenting

cell

VISTA

VIST

A

R-X

MDSC

Tumor associated

macrophage

R-X

VISTA Li et al. (Wang) Sci Rep 2017

Blando et al. (Sharma) PNAS 2018 Liru et al. Oncoimmun 2018

Deng et al. CIR 2019

Mulati et al. BJC 2018

VISTA can be expressed on tumor cells

or infiltrating immune cells in kidney cancer.

Mahoney, Novak, Freeman, Signoretti

VISTA is a negative immune checkpoint,

but its receptor on lymphocytes is not well established.

VSIG3 (IgSF11) Yang et al. J Biotecnol 2017

Wang et al. Immun 2018

VSIG8 Patent: WO 2016/090347 A1

PSLG1 Patent: WO 2018/169993 A1

Confirmed Binding at neutral pH

-

-

?

Y

Y

IgC domain

PD-L1 VISTA

VISTA has homology to the IgV domain of PD-L1,

but VISTA’s IgV is histidine rich.

IgV domain

At pH of 7, histidine residues

are neutral.

At acidic pH (~5-6),

histidine residues become

positively charged.

Korman et al. (BMS), Patent: WO 2018/169993 A1

PSGL-1 is expressed on lymphocytes

and is an negative immune checkpoint.

Tinoco et al. Immunity 2016

PSGL-1 is a negatively charged protein;

thus an interesting candidate receptor for VISTA.

VSIG3 (IgSF11) Yang et al. J Biotecnol 2017

Wang et al. Immun 2018

VSIG8 Patent: WO 2016/090347 A1

PSLG1 Patent: WO 2018/169993 A1

Confirmed Binding at neutral pH

-

-

?

Receptor

VISTA may also function as a receptor.

31 amino acids 96 amino acidsY

Y

IgC domain IgV domain

PD-L1

Unclear function ?

VISTA has homology to the IgV domain of PD-L1,

but a longer cytoplasmic tail.

IgV domain

VISTA

VISTA Antibodies to test the effect on VISTA function.

1:50

1:100

1:200

1:400

1:800

1:1600

1:3200

1:6400

1:12800

1:25600

1:51200

1:102400

0

50000

100000

150000

200000

250000

Dilution

PE

-A M

ean

300-mVISTA

#1 on mVISTA

#2 on mVISTA

#1 on hCD161

#2 on hCD161

#3 on hCD161

To test in functional in vitro assays:

proliferation assays,

cytotoxic assays,

phagocytosis assays

and in vivo orthotopic RENCA tumor growth.

The mouse kidney tumor RENCA

is a model of PD-1 resistant, VISTA expressing tumor.

RENCA is also a high myeloid: low

lymphocyte model of cancer

to test VISTA antibodies.

post injection:

Day 21 Day 28

RENCA is a locally

aggressive mouse

kidney tumor.

VS

IG8

VS

IG3

VIS

TA

PD

-1

PD

-L1

PD

-L2

Characterizing VISTA receptor:ligand function may lead

to defining tumors by actionable pathway.

Mahoney, Obi, Culhane

Mostly

specimens

from

patients

with clear

cell RCC

MIXED

papillary and

ccRCC

Mostly

papillary

RCC

Mostly

Chromophobe

Summary and Project Plan:

• VISTA is a negative immune checkpoint expressed in kidney

cancer.

• VISTA’s receptor on lymphocytes is not well established.

• The VISTA:Receptor interaction may be pH dependent.

• Test VISTA antibodies in the RENCA model in vivo and in

lymphocytes and macrophage functional assays in vitro.

Acknowledgements

Gordon Freeman

Xia Bu

Ye Gan

Baogong Zhu

Ping Hua

William Kaelin

Abhishek Chakraborty

Sabina Signoretti

Paul Catalano

Aedin Culhane

Rupal Bhatt

David F McDermott

Toni Choueiri

Glenn Bubley

DOD Early Career Investigator Idea Development Award, KC170162

Kidney SPORE Career Enhancement Program, P50CA101942

ASCO Young Investigator Award, supported by the Kidney Cancer Association, 2014

AACR Anna D. Barker Fellowship in Basic Cancer Research, 2014

Barr Award for Innovative Cancer Research, 2014