免疫检查点分子转化生长因子β受体II（TGFβRII）和T细胞免疫球蛋白粘膜3（TIM3）已被确认为前列腺癌T细胞免疫抑制的贡献因素。本研究的目标是通过同时靶向TIM3和TGFβRII来提高特异性前列腺膜抗原（PSMA）嵌合抗原受体T（CAR-T）细胞的肿瘤杀伤能力。为生成dnTGFβRII-trTIM3-PSMA-CAR-T（DT-PSMA-CAR-T）细胞，PSMA-CAR-T细胞的表面过表达了显性负性TGFβRII（dnTGFβRII）和截短外溶核TIM3（trTIM3）。通过体外杀伤实验和动物实验评估了DT-PSMA-CAR-T细胞的疗效。DT-PSMA-CAR-T细胞表现出能够消除PSMA阳性前列腺癌细胞的能力，即使在存在外源性TGF-β和/或TIM3激活抗体的情况下也能如此。此外，该细胞还展示了在体外移植了GAL9-PSMA-PC3细胞的免疫缺陷小鼠模型中消除肿瘤组织的能力，延长了生存时间且没有明显的毒性副作用。本研究强调，在T细胞表面上上调dnTGFβRII和trTIM3可能会降低TGFβRII和TIM3的抑制效应。版权所有 © 2023作者。由Elsevier B.V.出版。保留所有权利。

The immune checkpoint molecules, Transforming growth factor beta receptor II (TGFβRII) and T cell immunoglobulin and mucin domain 3 (TIM3), have been identified as contributors to T cell immune suppression in prostate cancer. The objective of this investigation was to improve the tumor killing capability of prostate-specific membrane antigen (PSMA)-chimeric antigen receptor T (CAR-T) cells by targeting TIM3 and TGFβRII simultaneously.To generate dnTGFβRII-trTIM3-PSMA-CAR-T (DT-PSMA-CAR-T) cells, the surface of PSMA-CAR-T cells was overexpressed with dominant negative TGFβRII (dnTGFβRII) and truncated extracellular TIM3 (trTIM3). The efficacy of DT-PSMA-CAR-T cells was assessed through in vitro killing experiments and animal experiments.The DT-PSMA-CAR-T cells demonstrated the ability to eradicate PSMA-positive prostate cancer cells, even in the presence of exogenous TGF-β and/or TIM3 activating antibodies. In addition, the cells demonstrated the ability to eliminate tumor tissue in an immunodeficient mouse model transplanted with GAL9-PSMA-PC3 cells in vitro, prolonging survival without significant toxic side effects.This study emphasizes that upregulating dnTGFβRII and trTIM3 on the surface of T cells can potentially diminish the inhibitory effects of TGFβRII and TIM3.Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.