USP7 targeting modulates anti-tumor immune response by reprogramming Tumor-associated Macrophages in Lung Cancer
USP7 targeting modulates anti-tumor immune response by reprogramming Tumor-associated Macrophages in Lung Cancer
复制标题
USP7靶向通过重编程肺癌中肿瘤相关巨噬细胞来调节抗肿瘤免疫反应
DOI：
10.7150/thno.47137
复制
发表时间：
2020-01-01
期刊：
影响因子：
12.4
通讯作者：
Yang, Kunyu
中科院分区：
文献类型：
--
作者：
Dai, Xiaomeng;Lu, Lisen;Yang, Kunyu
背景：肿瘤相关巨噬细胞（Tumor associated macrophages, tam）具有很强的可塑性，通过重编程可以清除肿瘤细胞，调节适应性免疫系统，用于肿瘤免疫治疗。去泛素化酶（deubiquitination酶，DUBs）可以从泛素修饰的底物中去除泛素（ubitin, Ub），它与致癌代谢有关，但不知道如何调节tam复极化。方法：采用逆转录- pcr方法检测巨噬细胞（MΦs）中DUB相关基因的表达。采用流式细胞术和免疫荧光法检测肿瘤微环境和脾脏免疫细胞的变化，包括M1 （CD11b+F4/80+CD86+CD206-）、M2 (CD11b+F4/80+CD86-CD206+) MΦs和IFN-γ+CD8+T细胞。增殖试验用于测定M2 MΦs经USP7抑制剂处理后对T细胞增殖的影响。Western blotting检测USP7的表达和MAPK通路的激活情况。利用TGCA数据库评估USP7在人肺腺癌（LUAD）免疫微环境中的作用。结果：筛选到51个DUB基因，发现USP7在M2中高表达，而在M1中不高表达MΦs。使用siRNA或USP7抑制剂特异性沉默USP7可导致M2 MΦs的表型和功能变化，有利于CD8+T细胞的体外增殖。USP7抑制剂延缓Lewis肺癌小鼠肿瘤生长，促进M1 MΦs和IFN-γ+CD8+T细胞的肿瘤浸润。tam的消耗减弱了这些治疗效果。USP7抑制被证明通过激活p38 MAPK通路介导MΦs重编程。施用USP7抑制剂可增加肿瘤中程序性细胞死亡配体1 （PD-L1）的表达，而阻断程序性细胞死亡蛋白1 （PD-1）可提供有效的抗肿瘤应答。临床资料显示，LUAD中USP7的高表达与先天免疫和适应性免疫呈负相关。结论：综上所述，这些结果提供了证据，表明针对USP7的治疗方法，结合免疫治疗，应考虑用于肺癌治疗。
Background: Tumor associated macrophages (TAMs) have strong plasticity and if reprogrammed, can clear tumor cells and regulate the adaptive immune system for cancer immunotherapy. Deubiquitinating enzymes (DUBs), which can remove ubiquitin (Ub) from Ub-modified substrates, have been associated with oncogenic metabolism but are not well-known for regulating TAMs repolarization. Methods: The expression of DUB related genes in macrophages (MΦs) was detected by reverse transcription-PCR. Flow cytometry and immunofluorescence were used to detect the changes of immune cells in the tumor microenvironment and spleen, including M1 (CD11b+F4/80+CD86+CD206-), and M2 (CD11b+F4/80+CD86-CD206+) MΦs, and IFN-γ+CD8+T cells. A proliferation assay was used to determine the effect of M2 MΦs treated with a USP7 inhibitor on T cell proliferation. Western blotting was used to detect the expression of USP7 and the activation of the MAPK pathway. The TGCA database was used to assess the role of USP7 in the immune microenvironment of human lung adenocarcinoma (LUAD). Results: 51 DUB genes were screened and USP7 was identified as a highly expressed gene in M2 but not M1 MΦs. Specific silencing of USP7 using siRNA or USP7 inhibitors led to phenotypical and functional changes in M2 MΦs, favoring CD8+T cells proliferation in vitro. USP7 inhibitors delayed tumor growth in mice with Lewis lung carcinoma, and promoted tumor infiltration of M1 MΦs and IFN-γ+CD8+T cells. Depletion of TAMs attenuated these therapeutic effects. USP7 inhibition was shown to mediate MΦs reprogramming by activating the p38 MAPK pathway. Administration of USP7 inhibitors increased the expression of programmed cell death ligand 1 (PD-L1) in tumors, while blocking programmed cell death protein 1 (PD-1) provided an effective anti-tumor response. Clinical databases suggest that high expression of USP7 in LUAD was negatively correlated with innate and adaptive immunity. Conclusions: Taken together, these results provide evidence to suggest that therapeutic approaches targeting USP7, in combination with immunotherapy, should be considered for lung cancer treatment.