Department of General Surgery, The Second Affiliated Hospital of Soochow University, Suzhou 215004, Jiangsu Province, China; Department of General Surgery, The Second Affiliated Hospital of Xuzhou Medical University, Xuzhou 221000, Jiangsu Province, China.
Department of General Surgery, The Second Affiliated Hospital of Xuzhou Medical University, Xuzhou 221000, Jiangsu Province, China.
Int Immunopharmacol. 2024 Jun 30;135:112317. doi: 10.1016/j.intimp.2024.112317. Epub 2024 May 25.
Colorectal cancer (CRC) is a significant global health challenge, with increasing rates of incidence and mortality. Despite advancements in immunotherapy, resistance, particularly due to T cell exhaustion, remains a major hurdle. This study explores the role of YWHAH, mediated by N4-acetylcytidine (ac4C) modification, in CRC progression and its impact on CD8 T cell exhaustion. Analysis of five paired CRC patient tissue samples using acetylated RNA immunoprecipitation and sequencing (acRIP-seq)identified ac4C-modified mRNAs. Functional assays, including cell culture, transfection, qRT-PCR, and immune assays, investigated the influence of YWHAH expression on CRC advancement. Bioinformatics analysis of TCGA data assessed the correlation between YWHAH and immune responses, as well as checkpoint inhibitors. Flow cytometry and Immunohistochemistry validated these findings, complemented by a co-culture experiment involving CD8 T cells and CRC cell lines (LOVO and HCT116). acRIP-seq revealed YWHAH as a potential driver of CRC progression, exhibiting ac4C modification-mediated stability and upregulation. High YWHAH levels correlated with adverse outcomes and immune evasion in CRC patients, showing strong associations with immune checkpoint proteins and modest correlations with CD8 T cell infiltration. Co-culture experiments demonstrated YWHAH-induced CD8 T cell exhaustion, characterized by decreased proliferation and increased exhaustion markers. NAT10-mediated ac4C modification enhanced YWHAH stability in CRC. The involvement of YWHAH in CD8 + T cell exhaustion suggests its potential as a therapeutic target and prognostic marker in CRC immunotherapy, highlighting the intricate interplay between epitranscriptomic modifications, the tumor microenvironment, and immune responses in CRC progression.
结直肠癌（CRC）是一个重大的全球健康挑战，其发病率和死亡率呈上升趋势。尽管免疫疗法取得了进展，但耐药性，尤其是由于 T 细胞耗竭，仍然是一个主要障碍。本研究探讨了 YWHAH 通过 N4-乙酰胞苷（ac4C）修饰在 CRC 进展中的作用及其对 CD8 T 细胞耗竭的影响。使用乙酰化 RNA 免疫沉淀和测序（acRIP-seq）分析了五个配对的 CRC 患者组织样本，鉴定了 ac4C 修饰的 mRNAs。功能分析，包括细胞培养、转染、qRT-PCR 和免疫分析，研究了 YWHAH 表达对 CRC 进展的影响。TCGA 数据分析评估了 YWHAH 与免疫反应以及检查点抑制剂之间的相关性。流式细胞术和免疫组织化学验证了这些发现，并通过涉及 CD8 T 细胞和 CRC 细胞系（LOVO 和 HCT116）的共培养实验进行了补充。acRIP-seq 显示 YWHAH 是 CRC 进展的潜在驱动因素，表现出 ac4C 修饰介导的稳定性和上调。高 YWHAH 水平与 CRC 患者的不良结局和免疫逃避相关，与免疫检查点蛋白具有强烈关联，与 CD8 T 细胞浸润具有适度相关性。共培养实验表明 YWHAH 诱导 CD8 T 细胞耗竭，表现为增殖减少和耗竭标志物增加。NAT10 介导的 ac4C 修饰增强了 CRC 中 YWHAH 的稳定性。YWHAH 参与 CD8 + T 细胞耗竭表明其在 CRC 免疫治疗中作为治疗靶点和预后标志物的潜力，突出了 CRC 进展中表转录组修饰、肿瘤微环境和免疫反应之间的复杂相互作用。
