Neuroimmune regulation of neurotropic JC virus by SF2/ASF in glial cells

神经胶质细胞中 SF2/ASF 对嗜神经 JC 病毒的神经免疫调节

• 批准号: 8660281
• 负责人: Ilker Kudret Sariyer
• 金额: $ 38.25万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-01 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8660281
• 关键词: Acquired Immunodeficiency Syndrome; Address; Alternative Splicing; Astrocytes; Autoimmune Diseases; Biochemical; Biological; Biological Assay; Biological Response Modifiers; Brain; Cell Culture Techniques; Cells; Characteristics; Childhood; Communication; Cytoplasm; Demyelinating Diseases; Disease; Down-Regulation; Evaluation; Gene Expression; Genetic Transcription; Goals; Human; Immune; Immune system; Immunocompromised Host; Incidence; Individual; Infection; JC Virus; Life; Life Cycle Stages; Mediating; Modeling; Molecular; Multiple Sclerosis; Neuraxis; Neuroglia; Oligodendroglia; Outcome; Patients; Peripheral; Polyomavirus; Population; Progressive Multifocal Leukoencephalopathy; Proteins; RNA Splicing; Regulation; Reporter Genes; Rest; Role; Sampling; Series; Signal Pathway; Staging; Testing; Therapeutic Intervention; Tissues; Viral; Viral Genes; Viral Genome; Viral Proteins; Virus; Virus Diseases; Virus Latency; cell type; cytokine; enhancing factor; high risk; latent infection; neurotropic; novel; novel strategies; operation; overexpression; promoter; research clinical testing; stable cell line; white matter

DESCRIPTION (provided by applicant): Progressive multifocal leukoencephalopathy (PML) is a fatal demyelinating disease caused by human neurotropic JC virus. JCV infects more than 80% of human population during the childhood and establishes a latent infection for the rest of the life. The virus reactivates under immunomodulatory conditions with an unknown mechanism, resulting in productive infection of oligodendrocytes and astrocytes in the central nervous system (CNS). JCV replicates almost exclusively in glial cells, and its promoter sequence, which has tissue- specific characteristics, tightly modulates expression of viral genome in appropriate cell types and immune conditions through communication with cellular factors. We recently identified the alternative splicing factor, SF2/ASF, as a potential regulator of JCV as its overexpression in glial cells strongly suppresses viral gene expression and replication. Unexpectedly, down-regulation of JCV by SF2/ASF is mediated at transcriptional stage, thus ascribing a novel role for SF2/ASF in the control of promoter activity. Results from our preliminary studies suggest that immune mediators secreted by PBMCs induce the expression of SF2/ASF, and inhibit the replication of JCV. These observations suggest operation of a novel immune signaling pathway between peripheral immune cells and glial cells that controls the immediate early stage of JCV gene expression during the course of viral reactivation. Our preliminary studies from JCV-infected glial cells also revealed a significant role of agnoprotein, a small viral protein crucial for JCV replication, in regulation of SF2/ASF. Expression of SF2/ASF is induced, and colocalized with agnoprotein in the cytoplasm of infected cells. Results from biochemical assays showed interaction of agnoprotein with SF2/ASF suggesting a functional cross-association between these two proteins. Results from reporter gene assays revealed that agnoprotein induces the viral transcription silenced by SF2/ASF. These observations provided us a rationale to study the role of SF2/ASF in JCV life cycle, and led us to formulate our central hypothesis which s that neuroimmune regulation of JCV replication is mediated by SF2/ASF, and viral agnoprotein interferes with this regulation. We propose to examine our hypothesis by (i) examining immune mediated regulation of SF2/ASF and replication of JCV during the latent as well as productive period of the viral infection and (ii) investigating the functional interplay between SF2/ASF and JCV agnoprotein in this regulation. We will employ molecular, cellular, and virological approaches to address these questions and examine the biological relevance of our findings by immunohistochemical evaluation of clinical samples from patients with PML disease. The information from these studies will offer a novel strategy for the treatment of PML.

进行性多灶性脑白质病（PML）是一种由嗜神经型JC病毒引起的致死性脱髓鞘疾病。儿童期感染超过80%的人口，并形成终身潜伏感染。该病毒在免疫调节条件下以未知机制重新激活，导致中枢神经系统（CNS）少突胶质细胞和星形胶质细胞的多产性感染。JCV几乎只在神经胶质细胞中复制，其启动子序列具有组织特异性，通过与细胞因子的通信，在适当的细胞类型和免疫条件下紧密调节病毒基因组的表达。我们最近发现了另一种剪接因子SF2/ASF作为JCV的潜在调节因子，因为它在神经胶质细胞中的过表达强烈抑制病毒基因的表达和复制。出乎意料的是，SF2/ASF在转录阶段介导了JCV的下调，因此SF2/ASF在控制启动子活性方面发挥了新的作用。