Temsirolimus Enhances Anti-Cancer Immunity by Inducing Autophagy-Mediated Degradation of the Secretion of Small Extracellular Vesicle PD-L1

SIMPLE SUMMARY: Immune checkpoint blockade therapies (ICBT) have increasing importance in patient survival and prognosis because it enhances immune cell activation by inhibiting the binding of programmed death-ligand 1 (PD-L1) of tumor and programmed death-1 (PD-1) of T cells. However, tumor-derived small extracellular vesicle (sEV) PD-L1 trigger low reactivity in immunotherapy because it promotes tumor growth and metastasis and inhibits activation of immune cell. In this study, temsirolimus (TEM) which the Food and Drug Administration (FDA) approved as a targeted anti-cancer drug, inhibited tumor-derived sEV PD-L1 secretion by activating autophagy. In addition, TEM induced systemic anti-cancer immunity by increasing the number and activation of CD4(+) and CD8(+) T cells. Therefore, TEM showed that the anti-cancer effect was better in the breast cancer-bearing-immunocompetent mice than in the nude mice. In summary, we suggest that TEM can overcome sEV PD-L1-mediated immunosuppression in patients with cancer through activation of the immune system in the body by inhibiting tumor-derived sEV PD-L1. ABSTRACT: Tumor-derived small extracellular vesicle (sEV) programmed death-ligand 1 (PD-L1) contributes to the low reactivity of cells to immune checkpoint blockade therapy (ICBT), because sEV PD-L1 binds to programmed death 1 (PD-1) in immune cells. However, there are no commercially available anti-cancer drugs that activate immune cells by inhibiting tumor-derived sEV PD-L1 secretion and cellular PD-L1. Here, we aimed to investigate if temsirolimus (TEM) inhibits both sEV PD-L1 and cellular PD-L1 levels in MDA-MB-231 cells. In cancer cell autophagy activated by TEM, multivesicular bodies (MVBs) associated with the secretion of sEV are degraded through colocalization with autophagosomes or lysosomes. TEM promotes CD8(+) T cell-mediated anti-cancer immunity in co-cultures of CD8(+) T cells and tumor cells. Furthermore, the combination therapy of TEM and anti-PD-L1 antibodies enhanced anti-cancer immunity by increasing both the number and activity of CD4(+) and CD8(+) T cells in the tumor and draining lymph nodes (DLNs) of breast cancer-bearing immunocompetent mice. In contrast, the anti-cancer effect of the combination therapy with TEM and anti-PD-L1 antibodies was reversed by the injection of exogenous sEV PD-L1. These findings suggest that TEM, previously known as a targeted anti-cancer drug, can overcome the low reactivity of ICBT by inhibiting sEV PD-L1 and cellular PD-L1 levels.