Spatially Resolved and Quantitative Analysis of VISTA/PD-1H as a Novel Immunotherapy Target in Human Non–Small Cell Lung Cancer

Abstract

Purpose Determine the localized expression pattern and clinical significance of VISTA/PD-1H in human NSCLC. Experimental Design Using multiplex quantitative immunofluorescence (QIF), we performed localized measurements of VISTA, PD-1 and PD-L1 protein in 758 stage I-IV NSCLCs from 3 independent cohorts represented in tissue microarray format. The targets were selectively measured in cytokeratin+ tumor epithelial cells, CD3+ T-cells, CD4+ T-helper cells, CD8+ cytotoxic T-cells, CD20+ B-lymphocytes and CD68+ tumor-associated macrophages. We determined the association between the targets, clinico-pathological/molecular variables and survival. Genomic analyses of lung cancer cases from TCGA was also performed. Results VISTA protein was detected in 99% of NSCLCs with a predominant membranous/cytoplasmic staining pattern. Expression in tumor and stromal cells was seen in 21% and 98% of cases, respectively. The levels of VISTA were positively associated with PD-L1, PD-1, CD8+ T-cells and CD68+ macrophages. VISTA expression was higher in T-lymphocytes than in macrophages; and in cytotoxic T-cells than in T-helper cells. Elevated VISTA was associated with absence of EGFR mutations and lower mutational burden in lung adenocarcinomas. Presence of VISTA in tumor compartment predicted longer 5-year survival. Conclusions VISTA is frequently expressed in human NSCLC and shows association with increased TILs, PD-1 axis markers, specific genomic alterations and outcome. These results support the immuno-modulatory role of VISTA in human NSCLC and suggests its potential as therapeutic target.

VISTA: Coming of age as a multi-lineage immune checkpoint

“…Similarly, VISTA expression is up‐regulated after ipilimumab (anti‐CTLA‐4) therapy in prostate cancer . In addition, VISTA expression seems to correlate with PD‐1/PD‐L1 expression; for example, in colon cancer, NSCLC and gastric cancer . Overall, these data on the expression of VISTA in human cancer are highly supportive that targeting of VISTA as part of immunotherapeutic strategies may be of great value.…”

“…This has been demonstrated in gastric cancer , ovarian cancer , colorectal cancer and primary oral squamous cell mesothelioma . Interestingly, in non‐small‐cell lung cancer (NSCLC) and hepatocellular carcinoma (HCC), tumor cell expression appears to be associated with improved objective survival (OS) . Currently, it is not clear if and how VISTA expression induced in CD45‐negative cells might be supportive for anti‐tumor immunity.…”

Expression and Significance of Immune Checkpoints in Clear Cell Carcinoma of the Uterine Cervix

“…Samples were considered positive for B7-H3 and B7-H4 when ≥5% of the TCs expressing these proteins at any intensity; the 5% cut-off was chosen based on a previous publication [15]. As VISTA expression on TCs and tumourassociated immune cells (TAICs) may have distinct prognostic values [16][17][18], we evaluated the expression of VISTA on these cell types separately. TCs and TAICS were each considered VISTA-positive when ≥5% of them were stained for this protein, as described in previous publications [17,18].…”

VISTA expression is associated with a favorable prognosis in patients with high-grade serous ovarian cancer

“…These studies suggest that VISTA may modulate a novel immune evasion mechanism and is thus a potential target for cancer immunotherapy. VISTA expression in human cancers has been reported in non-small cell lung cancer (NSCLC), hepatocellular carcinoma, colorectal carcinoma, oral squamous cell carcinoma, gastric carcinoma, acute myeloid leukemia, and gestational trophoblastic neoplasia [13][14][15][16][17][18][19][20][21]. Mulati et al also found that VISTA was highly expressed in human ovarian and endometrial cancers [20].…”