Wang Mei-Rong, Bai Cheng-Si, Dai Jian-Wei, Yang Lan, Quan Fang-Yi, Ma Jian-Chun, Chen Xing-Yuan, Zhu Shao-Wei, Xu Ying-Qi, Xiang Zhou-Fu, Jiang Ya-le, Cheng Qi, Zhang Wei-Hao, Chen Ke-Han, Wang Jian-Hua, Feng Yong, Chen Xiao-Ping, Xiong Yong, Chen Shu-Liang, Hou Wei, Xiong Hai-Rong
State Key Laboratory of Virology and Biosafety /Department of Infectious Diseases /Hubei Province Key Laboratory of Allergy and Immunology, School of Basic Medical Sciences /Zhongnan Hospital, Wuhan University, Wuhan, Hubei Province, China; Department of Clinical Laboratory, Yantai Yuhuangding Hospital, Yantai, Shandong Province, China.
State Key Laboratory of Virology and Biosafety /Department of Infectious Diseases /Hubei Province Key Laboratory of Allergy and Immunology, School of Basic Medical Sciences /Zhongnan Hospital, Wuhan University, Wuhan, Hubei Province, China.
Mol Cell Proteomics. 2025 May 23;24(7):100997. doi: 10.1016/j.mcpro.2025.100997.
Long noncoding RNAs (lncRNAs) are effective regulators of both RNA and protein functions throughout cell biology, including viral replication. Emerging studies have shown that lncRNAs activate or inhibit the replication and latency of HIV-1 by regulating different cellular mechanisms. Metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) is an oncogenic lncRNA required for paraspeckle integrity and has been proven to be linked to viral infection. However, the mechanisms by which it influences HIV-1 infection in macrophages remain unclear. In this study, we performed RNA-deep sequencing to compare the profiles of lncRNAs in macrophages with or without HIV-1 and found that MALAT1 was dramatically upregulated in HIV-1-infected macrophages. MALAT1 knockdown inhibited HIV-1 infection, whereas MALAT1 overexpression enhanced viral replication, indicating that MALAT1 promotes HIV-1 replication. We further performed proteomics analysis and found that coiled-coil-helix-coiled-coil-helix domain-containing 2 (CHCHD2) was the most downregulated protein affected by RNAi-mediated knockdown of MALAT1. We next demonstrated that MALAT1 favored HIV-1 replication in a CHCHD2-dependent manner and functioned as a competing endogenous RNA to regulate CHCHD2 expression by sponging miR-145-5p, which could mutually bind the MALAT1 and 3'UTR of chchd2 mRNA. Furthermore, knockdown of endogenous MALAT1 or CHCHD2 with specific small interfering RNAs (siRNAs) promoted the expression of IRF7, and enhanced the promoter activities of interferons-α and -β, increasing their production as well as that of a critical interferon-stimulated gene (ISG), myxovirus resistance protein B (MxB). Moreover, MALAT1 or CHCHD2 knockdown promoted the expression of STAT2 to enhance the production of downstream MxB, which expanded the role of CHCHD2 as a negative regulator of the innate immune response. These findings improve our understanding of MALAT1/miR-145-5p/CHCHD2 pathway regulation of HIV-1 replication in macrophages, providing new insights into potential targeted therapeutic interventions.
长链非编码RNA（lncRNAs）是细胞生物学中RNA和蛋白质功能的有效调节因子，包括病毒复制。新兴研究表明，lncRNAs通过调节不同的细胞机制来激活或抑制HIV-1的复制和潜伏。转移相关肺腺癌转录本1（MALAT1）是一种致癌lncRNA，是副斑点完整性所必需的，并且已被证明与病毒感染有关。然而，其影响巨噬细胞中HIV-1感染的机制仍不清楚。在本研究中，我们进行了RNA深度测序，以比较有或没有HIV-1的巨噬细胞中lncRNAs的谱，发现MALAT1在HIV-1感染的巨噬细胞中显著上调。MALAT1敲低抑制HIV-1感染，而MALAT1过表达增强病毒复制，表明MALAT1促进HIV-1复制。我们进一步进行了蛋白质组学分析，发现含卷曲螺旋-螺旋-卷曲螺旋结构域2（CHCHD2）是受RNAi介导的MALAT1敲低影响最大的下调蛋白。接下来，我们证明MALAT1以CHCHD2依赖的方式促进HIV-1复制，并作为竞争性内源RNA通过海绵化miR-145-5p来调节CHCHD2表达，miR-145-5p可与chchd2 mRNA的MALAT1和3'UTR相互结合。此外，用特异性小干扰RNA（siRNAs）敲低内源性MALAT1或CHCHD2可促进IRF7的表达，并增强干扰素-α和-β的启动子活性，增加它们以及关键的干扰素刺激基因（ISG）、黏液病毒抗性蛋白B（MxB）的产生。此外，MALAT1或CHCHD2敲低促进STAT2的表达以增强下游MxB的产生，这扩展了CHCHD2作为先天免疫反应负调节因子的作用。这些发现增进了我们对MALAT1/miR-145-5p/CHCHD2通路调节巨噬细胞中HIV-1复制的理解，为潜在的靶向治疗干预提供了新见解。
