Division of Dermatology, Department of Surgery, City of Hope National Medical Center, Duarte, California, USA; Beckman Research Institute, City of Hope National Medical Center, Duarte, California, USA.
Irell and Manella Graduate School of Biological Sciences, Beckman Research Institute, City of Hope National Medical Center, Duarte, California, USA.
J Invest Dermatol. 2022 Mar;142(3 Pt A):603-612.e7. doi: 10.1016/j.jid.2021.08.447. Epub 2021 Nov 11.
Cutaneous T cell lymphoma (CTCL) is characterized by a background of chronic inflammation, where malignant CTCL cells escape immune surveillance. To study how microRNAs (miRs) regulate T-cell exhaustion, we performed miR sequencing analysis, qRT-PCR, and in situ hybridization on 45 primary CTCL samples, three healthy skin samples, and CTCL cell lines, identifying miR-155-5p, miR-130b-3p, and miR-21-3p. Moreover, miR-155-5p, miR-130b-3p, and miR-21-3p positively correlated with immune checkpoint gene expression in lesional skin samples and were enriched in the IL-6/Jak/signal transducer and activator of transcription signaling pathway by gene set enrichment analysis. Further gene sequencing analysis showed decreased mRNA expression of the major negative regulators of Jak/signal transducer and activator of transcription signaling: SOCS, PIAS, and PTPN. Transfection of MyLa and HuT78 cells with anti-miR-155-5p, anti‒miR-21-3p, and anti‒miR-130b revealed a considerable increase in SOCS proteins along with a significant decrease in the levels of activated signal transducer and activator of transcription 3 and immune checkpoint surface protein expression as well as decreased cell proliferation. Downregulation of miR-155, miR-130, and miR-21 in CTCL cell lines decreased CTCL cell growth and facilitated CD8 T-cell-mediated cytotoxic activity, with concordant production of IFN-γ and CD107a expression. Our results describe the mechanisms of miR-induced T-cell exhaustion, which provide a foundation for developing synthetic anti-miRs to therapeutically target the tumor microenvironment in CTCL.
皮肤 T 细胞淋巴瘤 (CTCL) 的特征是慢性炎症背景，其中恶性 CTCL 细胞逃避免疫监视。为了研究 microRNAs (miRs) 如何调节 T 细胞衰竭，我们对 45 个原发性 CTCL 样本、3 个健康皮肤样本和 CTCL 细胞系进行了 miR 测序分析、qRT-PCR 和原位杂交，鉴定出 miR-155-5p、miR-130b-3p 和 miR-21-3p。此外，miR-155-5p、miR-130b-3p 和 miR-21-3p 在病变皮肤样本中与免疫检查点基因表达呈正相关，并且通过基因集富集分析在 IL-6/Jak/信号转导和转录激活因子信号通路中富集。进一步的基因测序分析显示 Jak/信号转导和转录激活因子主要负调节剂的 mRNA 表达减少：SOCS、PIAS 和 PTPN。用抗 miR-155-5p、抗 miR-21-3p 和抗 miR-130b 转染 MyLa 和 HuT78 细胞，发现 SOCS 蛋白表达显著增加，同时激活的信号转导和转录激活因子 3 和免疫检查点表面蛋白表达水平显著降低，细胞增殖减少。CTCL 细胞系中 miR-155、miR-130 和 miR-21 的下调降低了 CTCL 细胞的生长，并促进了 CD8 T 细胞介导的细胞毒性活性，同时伴有 IFN-γ 和 CD107a 表达的协同产生。我们的结果描述了 miR 诱导的 T 细胞衰竭的机制，为开发合成抗 miR 来治疗性靶向 CTCL 肿瘤微环境提供了基础。
