长链非编码RNA SNHG1调控记忆性CD8 T细胞分化及其机制研究

Memory CD8 T-cell plays a key role in protecting host from intracellular pathogens and cancer. While the immune functions of lncRNAs in regulating an array of immune responses have been established, whether they are involved in regulating memory CD8 T cell differentiation remains unclear. The applicant focused on immunogenicity of viruses as a Ph.D. student, and continue to work with Dr. Lilin Ye on memory CD8 T cell differentiation. Our preliminary data by profiling transcriptomes with RNA-Sequencing have indicated that the lncRNA SNHG1 was able to regulate memory CD8 T cell differentiation during acute viral infection. Knock-down of SNHG1 significantly reduced the formation of memory CD8 T cell pool. Furthermore, several potential binding proteins of SNHG1 were identified by RNA-pull down-MS. Next, we set out to more comprehensively investigate the regulation function of SNHG1 in memory CD8 T cell differentiation and elucidate the underlying molecular mechanism by which SNHG1-binding partner regulates memory CD8 T cell formation. This study will facilitate us to have a better understanding of memory CD8 T cell differentiation and help us improve the efficacies of immunotherapies and vaccines.

记忆性CD8 T细胞在抗病毒、胞内菌感染及肿瘤免疫中起关键作用。长链非编码RNA(lncRNA)被发现参与调控一系列免疫相关过程,但其是否调控记忆性CD8 T细胞的分化及机制并不清楚。申请人博士期间从事病毒免疫原性分析,博士后于叶丽林团队开展CD8 T细胞免疫记忆的相关研究。前期利用RNA-Seq技术,我们筛选到记忆性CD8 T细胞特异性表达的lncRNA SNHG1;将其敲低显著影响记忆性CD8 T细胞的形成,并进一步由RNA-pull down-质谱鉴定若干SNHG1 的潜在结合蛋白。下一步,拟利用体内、体外模型,结合病毒特异性CD8 T细胞过继转移,验 证SNHG1促进记忆性CD8 T细胞形成的作用并阐明SNHG1-结合蛋白调控此过程的分子机制。本研究将加深对CD8 T细胞免疫记忆多层次调控机制的认识,并为通过靶向lncRNA来提升疫苗介导 的CD8 T细胞应答提供一定的理论基础。

记忆性CD8+ T细胞对于机体起到重要的作用，能有效地保护机体免遭病原体的再次侵害。诸多因素参与调控记忆CD8+ T细胞的分化和维持，但长链非编码（lncRNA）在这一过程中的作用尚不明确。在本研究中，我们首先通过RNA-seq高通量测序和RT-qPCR验证，发现SNHG1在初始和记忆CD8+ T细胞中相对高表达，而在效应CD8+ T细胞中相对低表达。随后，通过敲减SNHG1在CD8+ T细胞中的表达，我们发现SNHG1可促进记忆CD8+ T细胞的分化。接下来，通过RNA pull down和RIP等实验，我们找到了与SNHG1相互作用的囊泡运输蛋白Vps13D。我们进一步通过免疫荧光染色等实验，证明了SNHG1-Vps13D可通过促进CD127 (IL7R)的胞内转运而影响其在细胞膜上的表达。最后，通过ChIP、流式细胞染色、TCF-1过表达等实验，我们发现SNHG1-Vps13D是通过影响IL7-CD127信号通路来调控记忆CD8+ T细胞分化的。总之，我们首次发现lncRNA SNHG1可促进记忆CD8+ T细胞的分化，我们也揭示了SNHG1促进记忆CD8+ T细胞分化的分子机制，为靶向lncRNA治疗感染性疾病和疫苗设计提供新的思路及理论基础。