Beschreibung:
<jats:title>Abstract</jats:title>
<jats:p>Immunotherapy strategies for the treatment of melanoma have achieved impressive clinical outcomes over the past decade. Response rates to checkpoint blockade by PD-1 and CTLA-4 antibodies range from 15-40%, while in adoptive cell therapy using tumor infiltrating lymphocytes (TILs), anti-tumor response is observed in approximately 50%. However, the need to improve immunotherapies is evident as the majority of patients are unresponsive to treatment. Dysfunctional T-cells are thought to contribute to failed responses to checkpoint inhibition. As such we sought to investigate the ability of drugs targeting the epigenetic regulatory machinery as a means to alter T-cell function(s) and improve the anti-melanoma response. Here we demonstrate that the HDAC6 selective inhibitor ACY1215 disrupts mTORC signaling pathways in T-cells obtained from melanoma patients. Phosphorylation of mTOR, RAPTOR and the downstream molecules AKT, SGK1, PKCa and S6K were reduced on CD4 and CD8 T-cells after ACY1215 in vitro treatment (p&lt;0.05). The levels of the Th2 cytokines IL-4, IL-6, IL-10 generated by ACY1215-treated T-cells (p&lt;0.05) were also decreased. Similar results were achieved with an SGK1 inhibitor, in agreement with published data demonstrating SGK1 as a regulator of Th2 polarization. Since the mTOR/RAPTOR complex is known to be involved in determining T regulatory (Treg) function, the effects of ACY1215 on Tregs were evaluated. Treatment in vitro with ACY1215 decreased phosphorylated mTOR and RAPTOR in Tregs, and reduced the levels of FOXP3. In a functional suppression assay, ACY1215-treated Tregs displayed a reduced ability to impair proliferation of effector T-cells (Teff) compared to control (DMSO: 10% vs ACY1215: 25% Teff proliferation, p&lt;0.05). To explore whether HDAC inhibition during expansion of tumor infiltrating lymphocytes (TIL) for adoptive transfer would improve their quality and anti-tumor reactivity, TIL isolated from melanoma surgical biopsies were cultured in vitro with IL-2 and ACY1215. Treatment with ACY1215 led to an accumulation of central memory CD4 and CD8 TILs (p&lt;0.01 and p&lt;0.05, respectively), which was maintained even after rapid expansion with anti-CD3 and anti-CD28 stimulation in vitro. Similarly, ACY1215 treatment of T-cells derived from peripheral blood of melanoma patients and healthy donors also displayed an increased central memory phenotype, characterized by expression of CD45RO, CD62L and CCR7 (p&lt;0.05). Inhibition of AKT has been shown to increase T-cells with memory characteristics, and the use of an AKT inhibitor also resulted in accumulation of central memory T-cells. Confirming the observed phenotypic changes, microarray analysis of ACY1215-treated TILs revealed up-regulation of genes associated with a T-cell central memory and inflammatory response (e.g. SELL, LEF1, TNFRSF9) and downregulation of genes associated with Treg function (e.g. LGMN, CXCL8). Collectively these data suggest that reprogramming T-cells with epigenetic modulators may improve melanoma immunotherapy by reducing Treg suppression and production of immunosuppressive cytokines, while favoring generation of central memory T-cells.</jats:p>
<jats:p>Citation Format: Andressa L. Sodre, David M. Woods, Amod Sarnaik, Brian C. Betts, Jeffrey S. Weber. Epigenetic reprogramming of T-cells from metastatic melanoma patients enhances central memory and decreases Th2/Treg phenotypes [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr B109.</jats:p>