VISTA/PD-1H is an Ig domain-containing type I transmembrane protein able to suppress T-cell activation and is being evaluated as a candidate anti-cancer immunotherapy target. VISTA can bind P-selectin glycoprotein ligand-1 (PSGL1) and V-set and immunoglobulin domain containing 3 (VSIG3) suggesting these interactions could mediate VISTA's immunomodulatory effect. We studied the tissue distribution of VISTA, PSGL1 and VSIG3 in human non-small cell lung cancer (NSCLC).
We used multiplexed quantitative immunofluorescence (QIF) to simultaneously measure DAPI, cytokeratin (CK), VISTA, PSGL1 and VSIG3 in 48 paired tumor/normal lung samples and in 850 stage I-IV NSCLCs from four independent cohorts represented in tissue microarray format. The first 2 NSCLC cohorts (Cohorts #1, n=382 and #2, n=282) included cases treated with standard of care non-immunotherapy. Cohort #3 included 137 lung adenocarcinomas with analysis of mutant oncogenic drivers; and Cohort #4 included 49 NSCLC cases treated with PD-1 axis blockers. The targets were selectively measured in CK+ tumor cells and CK-negative stromal cells. We also determined the spatial association between VISTA and its binding partners by fluorescence signal co-localization. The association between target levels, clinicopathologic/molecular variables, tumor infiltrating lymphocytes (TILs), PD-L1 expression and survival was established.
PSGL1 was located predominantly in stromal/immune cells and VSIG3 was detected in both tumor and stromal compartments. Both targets were expressed at higher levels in NSCLC than in non-tumor lung tissue and showed a positive association with VISTA in cancer tissues. Using the visual detection threshold PSGL1 and VSIG3 expression were detected in >90% of cases from cohorts #1 and #2 and showed positive association with CD3+ TILs and PD-L1 levels. PSGL1 was higher in lung adenocarcinomas harboring EGFR mutations than in tumors with KRAS variants or cases lacking mutations in both oncogenes. Elevated VISTA/PSGL1 co-localization was significantly associated with longer 5-year overall survival in cases not treated with immunostimulatory therapy (Cohorts #1 and #2). However, an opposite association was seen in cases treated with PD-1 axis blockers, where elevated VISTA/PSGL1 co-expression was associated with shorter survival.
PSGL1 and VSIG3 are frequently expressed in human NSCLC. Expression of PSGL1 is associated with increased tumor immune infiltration and activating EGFR mutations. High baseline VISTA/PSGL1 co-expression is associated with adverse outcome after PD-1 axis blockers. The latter suggest VISTA/PSGL1 as a dominant immune evasion pathway independent from PD-1/PD-L1 axis in a subset of human NSCLC. Validation of these findings is ongoing.
Citation Format: Alicia Ding, Franz Villarroel-Espindola, Adam Ducler, Brian S. Henick, Shruti Desai, Nicole Gianino, Jon Zugazagoitia, David L. Rimm, Alain Robert, Francisco Cruzalegui, Pierre Ferré, Roy Herbst, Miguel Sanmamed, Lieping Chen, Kurt A. Schalper. VISTA/PSGL1 axis as a dominant immunomodulatory pathway in human non-small cell lung cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5525.