Regulation of NK cell antiviral responses through alternative splicing

通过选择性剪接调节 NK 细胞抗病毒反应

基本信息

批准号：
10388402
负责人：
Daniel Ramos Ram
金额：
$ 17.58万
依托单位：
BETH ISRAEL DEACONESS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-15 至 2023-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10388402
关键词：
Abnormal Cell Acute Affect Alternative Splicing Antigens Antisense Oligonucleotides Antiviral Response Area Cancerous Candidate Disease Gene Cell physiology Cells Cellular biology Characteristics Chronic Cryopreservation Cytomegalovirus Data Development Disease ES04 Effector Cell Epigenetic Process Event Evolution Exhibits Exons Family Family member Female Flow Cytometry Functional disorder Gene Expression Gene Expression Profile Gene Expression Regulation Genes Genetic Transcription Goals HIV HIV Infections HIV therapy Human Immune Immune response Immunity Immunology Impairment Inactivated Vaccines Individual Infection Infection Control Innate Immune Response Innate Immune System Laboratories Lead Macaca Macaca mulatta Malignant Neoplasms Mediating Memory Modification NUP214 gene Natural Killer Cells Output Pattern Peptides Peripheral Blood Mononuclear Cell Persons Play Population Property Protein Isoforms Publishing RNA RNA Splicing Regulation Reporting Research Resistance Role SIV Sampling Small Interfering RNA Spliced Genes TNFRSF6 gene Testing Therapeutic Intervention Third Generation Sequencing Tissues Training Transcript Vaccination Vaccines Variant Viral Viral reservoir Virus Virus Diseases Work acute infection adaptive immunity antiretroviral therapy chronic infection cohort environmental change epitranscriptomics gene product immunosenescence improved in vivo influenza virus vaccine insight interest male member novel peripheral blood post-doctoral training pre-doctoral protein complex receptor response stem transcription factor transcriptome transcriptome sequencing transcriptomics tumor vaccine response vaccine strategy

项目摘要

Project Summary/Abstract Natural killer (NK) cells have been shown to play a significant role in regulating viral infections as well as in eliminating cancerous cells. Traditionally, NK cells are classified as members of the innate immune system because of their ability to respond rapidly and non-specifically to infected or abnormal cells. Interestingly, burgeoning evidence suggests NK cells may also possess peptide-specific memory responses (previously only attributable to adaptive immunity), especially in the context of several viral infections including HIV/SIV and CMV. How NK cells can form these adaptive responses is the subject of an emerging area in immunology research, which also includes studying trained immunity of traditionally innate effectors like NK cells. Whether adaptive NK cell responses are the result of epigenetic modifications as observed in trained immunity, or the product of pre- formed polymorphic receptors like KIRs or members of the NKG2 family which interact with MHC molecules, or yet to be discovered receptors is to be determined. Here, we propose that investigating the NK cell transcriptome can provide insights into how infection or disease states can modulate NK cell function particularly by assessing how changes in alternative gene splicing following infection result in NK cell dysfunction or the formation of adaptive NK cell responses. Alternative splicing is a mechanism utilized by cells in order to produce multiple gene products from an individual gene, by splicing exons together in various combinations, sometimes leading to gene products with very different functions. Regulation of gene splicing is complicated, but it has been shown that environmental changes as are seen during viral infection or disease can gene splicing patterns. Using preliminary data from sorted NK cells in an acute SIV-infection cohort we have already identified several candidate alternatively spliced genes that exhibit differential isoform expression following SIV infection. Of particular interest are members of the bHLH transcription factor (TF) family due to their association with the formation of memory responses and NK cell development. Further, in a proof-of-concept study we have shown that NKp30 isoform expression is altered following ex vivo stimulation or even following vaccination in rhesus macaque and in human NK cell samples. Here, I propose to identify novel gene candidates and assess the roles of the bHLH TF splice variants and other candidate genes in rhesus macaque and human NK cells. Through this work I aim to determine whether specific gene isoform expression impacts normal NK cell functions or the formation of adaptive NK cell responses in vivo in rhesus macaques during acute and chronic SIV infection. The approaches outlined in this proposal will allow me to combine my postdoctoral training in NK cell biology with my predoctoral training assessing the role of alternative splicing in vivo in order to determine the roles that several gene candidates have on normal or adaptive NK cell responses during retroviral infection. Our ultimate goal is to identify and target specific isoforms to manipulate NK cell function in vivo for the purposes of improving current vaccine responses or eliciting targeted NK cell-specific therapies for HIV.

项目摘要/摘要天然杀手（NK）细胞已显示在调节病毒感染以及在消除癌细胞。传统上，NK细胞被归类为先天免疫系统的成员由于它们能够快速响应对感染或异常细胞的特异性反应。有趣的是，新兴的证据表明，NK细胞也可能具有特异性的记忆反应（以前仅归因于适应性免疫），特别是在包括HIV/SIV和CMV在内的几种病毒感染的背景下。 NK细胞如何形成这些适应性反应是免疫学研究中新兴领域的主题，这还包括研究传统上固有效应子（如NK细胞）的训练有素的免疫力。是否适应NK 细胞反应是在受过训练的免疫力中观察到的表观遗传修饰的结果，或与MHC分子相互作用的KIRS或NKG2家族的成员形成的多态受体，或然而，要确定被发现的受体。在这里，我们建议研究NK细胞转录组可以提供有关感染或疾病状态如何调节NK细胞功能的见解，特别是通过评估感染后替代基因剪接的变化如何导致NK细胞功能障碍或形成自适应NK细胞反应。替代剪接是单元使用的一种机制，以产生多个来自单个基因的基因产物，通过将外显子剪接在一起，有时是领先的到具有截然不同功能的基因产物。基因剪接的调节很复杂，但已显示在病毒感染或疾病期间看到的环境变化可以基因剪接模式。使用急性SIV感染队列中分类的NK单元的初步数据，我们已经确定了几个 SIV感染后表现出差异同工型表达的候选基因。的特别兴趣是BHLH转录因子（TF）家族的成员内存反应和NK细胞发展的形成。此外，在概念证明的研究中，我们已经显示在离体刺激甚至恒河猴疫苗接种后，NKP30同工型表达会改变猕猴和人类NK细胞样品。在这里，我建议识别新型基因候选者并评估角色 BHLH TF剪接变体和恒河猴和人类NK细胞中的其他候选基因的。通过这个我旨在确定特定基因同工型表达是否会影响正常NK细胞功能还是急性和慢性SIV感染过程中恒河猕猴中体内自适应NK细胞反应的形成。该提案中概述的方法将使我能够在NK细胞生物学中结合博士后培训通过我的遗传学训练，可以评估体内替代剪接的作用，以确定角色在逆转录病毒感染期间，几个基因候选物具有正常或适应性NK细胞反应。我们的最终目的是识别和靶向特定的同工型以在体内操纵NK细胞功能以改善的目的当前的疫苗反应或引起针对HIV的靶向NK细胞特异性疗法。