Anti-OX40 Antibody Directly Enhances The Function of Tumor-Reactive CD8⁺ T Cells and Synergizes with PI3Kβ Inhibition in PTEN Loss Melanoma.

Abstract

OX40 agonist-based combinations are emerging as a novel avenue to improve the effectiveness of cancer immunotherapy. To better guide its clinical development, we characterized the role of the OX40 pathway in tumor-reactive immune cells. We also evaluated combining OX40 agonists with targeted therapy to combat resistance to cancer immunotherapy.<b>Experimental Design:</b> We utilized patient-derived tumor-infiltrating lymphocytes (TILs) and multiple preclinical models to determine the direct effect of anti-OX40 agonistic antibodies on tumor-reactive CD8⁺ T cells. We also evaluated the antitumor activity of an anti-OX40 antibody plus PI3Kβ inhibition in a transgenic murine melanoma model (<i>Braf</i> mutant, PTEN null), which spontaneously develops immunotherapy-resistant melanomas. We observed elevated expression of OX40 in tumor-reactive CD8⁺ TILs upon encountering tumors; activation of OX40 signaling enhanced their cytotoxic function. OX40 agonist antibody improved the antitumor activity of CD8⁺ T cells and the generation of tumor-specific T-cell memory <i>in vivo</i>. Furthermore, combining anti-OX40 with GSK2636771, a PI3Kβ-selective inhibitor, delayed tumor growth and extended the survival of mice with PTEN-null melanomas. This combination treatment did not increase the number of TILs, but it instead significantly enhanced proliferation of CD8⁺ TILs and elevated the serum levels of CCL4, CXCL10, and IFNγ, which are mainly produced by memory and/or effector T cells. These results highlight a critical role of OX40 activation in potentiating the effector function of tumor-reactive CD8⁺ T cells and suggest further evaluation of OX40 agonist-based combinations in patients with immune-resistant tumors.