The Negative Checkpoint Regulator VISTA in Cancer and Tolerance
Isabelle Le Mercier, PhD Randolph Noelle Lab Presenter Disclosure Information
Isabelle Le Mercier
The following relationships exist related to this presentation:
In part supported by a grant from ImmuNext, Inc. and Janssen Biotech, Inc.
2 What are the Immune Checkpoints ? IG SUPERFAMILY TNF/TNFR SUPERFAMILY
B7-1/CD80 CTLA-4 B7-2/CD86 CD28 B7-1/CD80 OX40 OX40L PD-L1/B7-H1 PD-1
PD-L2/B7-DC RGMb CD40 CD154/CD40L B7-H2/ICOSL ICOS
B7-H3 CD27 CD70 APC B7-H4 T cell APC VISTA VISTA 4-1BB 4-1BBL
MHC TCR LAG3 GITR GITRL Gal-9 TIM-3 HVEM BTLA HVEM LIGHT CD226 PVR TIGIT VISTA (PD-1H, DD1a)
V-region of Immunoglobulin containing Suppressor of T cell Activation
Negative Checkpoint Regulator
• Membrane protein of the Ig SF • 92 Aa cytoplasmic domain • Homologous to PD-L1 (ligand) or PD-1 (receptor)
• Hematopoietically-restricted * VISTA is expressed on: • Myeloid cells • T cells • In the TME on MDSCs, human TILs
Controls inflammation through the suppression of T cell and myeloid cell mediated activities It is likely both a ligand and a receptor VISTA Biology VISTA is expressed on T cells and myeloid cells
100 Total CD4 CD8 CD11b high B cells spleen80 T cells T cells cells cells 60 Murine
% % of Max 40 spleen 20
0 10 0 10 1 10 2 10 3 10 4 VISTA
Human PBMCs
VISTA
Li Wang et al. J Exp Med 2011 Louise Lines et al. Cancer Res. 2014 VISTA is expressed on T cells and myeloid cells
Anti-VISTA on WT Anti-VISTA on KO Ham Ig on WT
Marginal Zone T cell zone SPLEEN
CD11b CD3 VISTA Over expression of VISTA on APCs inhibits T cell activation A20 B cell line + OVA peptide + OVA specific (D011) TCR Tg T cells
VISTA on APCs inhibits T cell proliferation VISTA on APCs inhibits activation markers upregulation A20-RFP A20-VISTA
CFSE
Li Wang et al. J Exp Med 2011 Jiannan Li Over expression of VISTA on tumor cell prevents antitumor immunity
2 weeks MCA105-VISTA MCA105-RFP Immunization Tumor challenge Irradiated tumor cells
Li Wang et al. J Exp Med 2011 Activated T cell and myeloid cell phenotype in VISTA deficient mice
Mild splenomegaly/lymphadenopathy in aged mice
Increased in myeloid populations
Gradual accumulation of spontaneously activated T cells (CD44+ CD62L low)
Enhanced T-cell responsiveness
Increased infiltration in specific organs
No systemic or organ specific autoimmune disease VISTA deficiency exacerbates EAE, model of MS
Myelin specific (2D2) TCR Tg WT Myelin specific (2D2) TCR Tg x VISTA KO - No disease - Spontaneous EAE disease (MS model) - Ascending paralysis - Reduced survival
Li Wang et al. PNAS 2014 VISTA deficiency on the host exacerbates passive EAE
Passive EAE, MS model Immune deficient
Myelin specific T cells Host KO Host KO Host WT T cells KO T cells WT T cells KO
HostHost WTWT TT cellscells WTWT Disease score Disease
Li Wang et al. PNAS 2014 VISTA deficiency in SLE prone mice exacerbates disease
Sle1/Sle3 WT Sle1/Sle3 x VISTA KO - Late incidence lupus - Early incidence lupus (starts 7 weeks) - Reduced survival (< 20 weeks)
100 500 B6 VISTA -/- 400 sle1,3 300 sle1,3xVISTA -/- 50 200 Protein (mg/dL) Protein 100 survival Percent 0 0 5 10 15 20 25 0 0 10 20 30 40 50 Weeks Weeks
Sabrina Ceeraz Delong
o
k
A
T
S
I V
**
6 B 0
750 500 250
1000 # IFNg spots in dLN in spots IFNg # eg function suppressive / conversion Allogeneic skin skin Graft Allogeneic 1 week Time (anti-CD40L) Costimulatory Blockade VISTA deficiency prevents allograft tolerance allograft prevents deficiency VISTA Donor Specific Transfusion Donor Transfusion Specific 0 20 40 60 80 0
75 50 25
100
Graft survival survival Graft Graft No defect in graft specific T cell deletion and Tr and deletion cell T specific graft in defect No Loss of VISTA from the host is responsible for enhanced graft T cell response
ELISPOT Recall
OVA specific T cells OVA skin Graft
2.0×10 4 ** *** Host WT host VISTA KO host 4 T cell 1.5×10
WT WT WT WT KO 1.0×10 4 T cell Normal Elevated response response 5.0×10 3 # IFNg spots in dLN in spots IFNg # KO WT KO KO VISTA KO 0.0 Normal Elevated T cell t t s s response response o o h h host T host W KO WT KO WT T cells KO T cells VISTA intrinsically regulates innate immunity
Listeria Spleen IV infection Bug (CFU) count
Early infection control Control in T cell deficient in VISTA KO RAG KO x VISTA KO
WT ** 6 KO 6 10 *** 10 10 5 10 5
10 4 10 4
10 3 10 3 CFU/spleen CFU / spleen / CFU
10 2 10 2 0 1 2 3 4 5 RAG Days post-infection
Louise Lines RAG x VISTA VISTA negatively regulates T cells and myeloid cells activities
Negatively regulates T cell responses
- acts as a ligand expressed on APCs (host)
Negatively regulates innate immunity
- acts as a receptor
Prevents autoimmunity in various suceptible models
Is critical for tolerance induction VISTA in Cancer VISTA is highly expressed in tumor infiltrating MDSCs
PD-L1 VISTA Melanoma B16 OVA Melanoma B16 OVA CD45 TILs Tumor cells Isotype
Bladder carcinoma Melanoma MB49 B16 OVA MDSCs CD11b hi CD11c - Gr1 +
TILs DAPI CD11b VISTA dLN CD11b+/VISTA+ cells Non dLN Isotype
VISTA Le Mercier et al. Cancer Res. 2014 Hypoxia upregulates VISTA expression in the TME
HRE1 HRE2 Bladder carcinoma MB49 ACGTG GCGTG CD45 TILs VISTA -1200 -800 -400
21% O 2 1% O 2
Pimonidazole
Gr1 Non hypoxic
Relative Luciferase Units Luciferase Relative Hypoxic Empty HRE1 Mut-HRE1 HRE2 Mut-HRE2 Isotype Vector
CD11b Jie Deng Rodwell Mabaera VISTA Anti -VISTA monotherapy controls tumor growth in multiple models
Melanoma Bladder carcinoma Fibrosarcoma B16-OVA MB49 MCA105
Le Mercier et al. Cancer Res. 2014 Anti -VISTA mediated changes in TME and antitumor immune responses
Melanoma B16-OVA
Le Mercier et al. Cancer Res. 2014 Anti -VISTA synergizes with anti -PD-1 to control tumor growth CT26 Colon Carcinoma
Early intervention (2-3 mm) Late intervention (5-6 mm)
Control Ig 1500 control 2500 ) ) anti-VISTA13F3 3 3 2000 anti-PD-1PD-1 1000 Combocombo 1500 co ntro l 1000 500 500 Tumor volume (mm volume Tumor Tumor volume (mm volume Tumor 0 0 0 10 20 30 5 7 1012141719212426293133 Days post-tumor inoculation Days post-tumor inoculation Combo : 50% CR Other groups 0% Louise Lines Anti-VISTA mediated changes in cancer immunity
VISTA is highly expressed in the TME: - Expression on MDSCs - Transcriptionally regulated by hypoxia
Anti-VISTA mediated myeloid changes: - Decrease in MDSCs - Increase in macrophages and activated DCs
Anti-VISTA mediated lymphoid changes: - Increase in T cells infiltration and recall responses
Anti-VISTA synergizes with anti-PD-1 Therapy VISTA is a potential target in lung cancer
Tumor cells
TILs VISTA regulation of immunity
VISTA is a negative regulator ligand of TUMOR T cell activation 1 Naïve/early activated Tumor T cell VISTA infiltrating MDSCMDSC VISTA T cell VISTA VISTA is a negative APC regulator receptor VISTA of T cell activation 3 VISTA is a receptor that regulates the biology of myeloid cells 2
1. Wang (J Exp Med 2011), Wang (PNAS 2014), Le Mercier (Cancer Res 2014); Lines (Cancer Immunol 2014); 2. Bharaj (PLoS One 2014) and unpublished data 3. Flies (J Clin Invest 2014), Flies (J Immunol 2015), Flies (J Immunol 2011) Geisel School of Medicine at Dartmouth
Janssen Pharmaceutical Companies of Johnson & Johnson
Randolph Noelle Team ImmuNext Rodwell Mabaera, M.D. Ph.D. Dave DeLucia Louise Lines, Ph.D. Jay Rothstein Sabrina Ceeraz Delong, Ph.D Catherine Carriere Anna Kuta, Ph.D. Tse Dan Elizabeth Nowak, Ph.D. Phil Gobeil Tommy Broughton David Blair Jie Deng Mike Molloy Yuchi Lee Yalin Guo Jiannan Li Xin Huang Aurelien Sarde Dongxu Son Petra Sergent Dov Pechenick Maria Day Rheumatology Ron Hirte Christopher Burns , M.D. Nick Schwertner Mike Nelson