Abstract 391: Lrg1 blockade modulates the tumor immune microenvironment and improves the efficacy of cancer therapeutics

Abstract

Abstract Although the use of checkpoint inhibitors and adoptive T cell therapy have been reported in multiple cancers resulting in a plethora of clinical trials, the efficacy of these therapeutic strategies is often limited by the presence of impaired vasculature in the tumour mass. Indeed, in solid tumors, blood vessels are dysfunctional, leaky and poorly perfused, and thwart not only the efficacy of systemically derived drugs but also the extravasation of T cells, promoting an immunosuppressive microenvironment. Leucine-rich alpha-2-glycoprotein 1 (LRG1) is a secreted glycoprotein that is associated with pathogenic neovascularization, autoimmune diseases, chronic inflammation, and is significantly up-regulated in many solid cancers. We have observed that treatment with a LRG1 function-blocking antibody (15C4) significantly limits growth of solid tumors inducing vessel normalization and functionality. In the present study, we aim to investigate if LRG1 blockade enhances an immunostimulatory milieu, making tumors more responsive to current immunotherapies. Murine melanoma cells harbouring the internal influenza nucleoprotein antigen, NP68, were grafted subcutaneously before antigen-specific F5B6 CD8+ T cells were also transferred. Interestingly, the combination with 15C4 promoted a greater reduction in tumor growth rate. Moreover, not only was the number of donor CD8+ T cells significantly higher when delivered with 15C4, but also the total number of CD3+ T cells was increased following combination therapy, suggesting a more robust host-derived lymphocyte trafficking. As blocking LRG1 normalizes the tumor vasculature and enhances T cell extravasation, we then evaluated whether 15C4 could improve delivery and efficacy of an anti-PD1 checkpoint inhibitor. As monotherapies, both 15C4 and anti-PD1 antibodies induced a marked reduction in tumor volume but when the two were combined, a significant additive effect was also observed. Histological analysis revealed that the combination therapy was associated with a significantly higher number of activated CD8+ T cells, together with enhanced expression of Granzyme B. Current studies are investigating whether the observed boost in cytotoxic T cell density and functionality is a direct consequence of vessel normalization or whether LRG1 blockade is able to modulate the tumour immune microenvironment at different levels: for example, FACS analysis of tumour-infiltrating leukocytes revealed a reduction in the percentage of regulatory T cells after dual therapy, while histological analysis suggested an increase in the number of M1 macrophages. These data show that LRG1 blockade orchestrates a shift in the tumour microenvironment from being immune silent to immune active and, therefore, might represent a novel tool to enhance the efficacy of cancer therapies. Citation Format: Carlotta Camilli, Camilla Pilotti, Marie N. O'Connor, David Kallenberg, Markella Alatsatianos, Angharad H. Watson, Julia Ohme, Athina Dritsoula, Ann Ager, Stephen E. Moss, John Greenwood. Lrg1 blockade modulates the tumor immune microenvironment and improves the efficacy of cancer therapeutics [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 391.