Role of Daxx in Reovirus-induced Neuronal Apoptosis

Daxx 在呼肠孤病毒诱导的神经元凋亡中的作用

基本信息

批准号：
8143541
负责人：
Kalen Ray Dionne
金额：
$ 2.71万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-15 至 2015-09-14
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8143541
关键词：
Adaptor Signaling Protein Address Annexins Antibodies Antiviral Response Antiviral Therapy Apoptosis Apoptotic Binding Biological Assay Brain Brain Diseases Brain Injuries Cause of Death Cell Death Cell membrane Cellular Immunity Central Nervous System Diseases Cessation of life Cytoplasm Data Development Diagnostic Disease Dominant-Negative Mutation Encephalitis Enzymes Etiology Family Fibroblasts Gene Expression Genes Goals Image Immune response Immunity Immunofluorescence Immunologic In Vitro Induction of Apoptosis Interferon Activation Interferons JUN gene Laboratories Lead Link MAPK14 gene Measures Mediating Mediator of activation protein Membrane Messenger RNA Methodology Mitogen-Activated Protein Kinases Modeling Molecular Morbidity - disease rate Mus N-terminal Neonatal Neurobiology Neurons Neuropathogenesis PC12 Cells Pathway interactions Patients Pharmacologic Substance Phosphorylation Phosphotransferases Play Process Protein Overexpression Proteins Receptor Activation Receptor Signaling Recombinant Interferon Relative (related person)Reoviridae Infections Reovirus Reovirus 3 Research Role Secondary to Serotyping Signal Pathway Signal Transduction Slice Small Interfering RNA Staining method Stains Study models Techniques Tissues Transfection Up-Regulation Viral Viral Encephalitis Virus Virus Diseases Western Blotting Wild Type Mouse Work arm base brain tissue caspase-3 cellular targeting human DAXX protein in vitro Model in vivo in vivo Model mortality nervous system disorder neuron apoptosis neuroprotection new therapeutic target novel overexpression response stress activated protein kinase stress-activated protein kinase 1 therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Neuronal apoptosis is a common pathological feature in neurological disorders of diverse etiology. Viral encephalitis, an exemplary disease associated with neuronal apoptosis, remains a significant cause of morbidity and mortality due to severely limited diagnostic and treatment options. The primary goal of the proposed research is to utilize in vivo, ex vivo, and in vitro models of reovirus infection to characterize signaling pathways activated during neuronal viral infection and, ultimately, to identify novel therapeutic targets for neuroprotection. Fas death receptor signaling and c-Jun N-terminal Kinase (JNK) activation are known to contribute to reovirus-induced neuronal apoptosis. These seemingly isolated findings are consolidated into a broader model, described in this proposal. As discovered in various non-viral models of apoptosis, Fas signaling originating at the cell membrane may initiate intracellular JNK activation by proceeding through the Fas-Daxx-ASK1-Sek1-JNK apoptotic pathway. I hypothesize that Daxx is an especially important mediator of apoptosis in the context of viral infection because its expression is controlled by the host anti-viral immune response. In Aim 1, models of dominant-negative Daxx overexpression (in vivo and in vitro) and siRNA- mediated Daxx knockdown (in vitro only) will be utilized to determine the relative importance of Daxx in reovirus-induced apoptosis and neuropathogenesis. Aim 2 will employ immunofluorescence imaging and Western blotting techniques to assess the binding interactions and signaling roles of Daxx following reovirus infection. Finally, Aim 3 will address the possibility that Daxx expression is regulated by interferon (IFN) released by the virally infected host. Overall, I hypothesize that IFN signaling and consequential Daxx gene expression leaves virally infected neurons vulnerable to apoptosis. The proposed studies will probe the interfaces between viral infection, immunity, and cellular/tissue/host death to deepen our understanding of the viral-host interaction and neuronal apoptotic signaling. Furthermore, these studies will potentially build the theoretic basis for the eventual development of neuroprotective pharmaceuticals.

描述（由申请人提供）：神经凋亡是多种病因的神经系统疾病中的常见病理特征。病毒性脑炎是一种与神经元凋亡相关的示例性疾病，由于严重有限的诊断和治疗选择，导致发病率和死亡率的重要原因。拟议的研究的主要目标是利用体内，体内和小葡萄病毒感染的体外模型来表征神经元病毒感染过程中激活的信号通路，并最终确定神经保护的新型治疗靶标。已知FAS死亡受体信号传导和C-JUN N末端激酶（JNK）激活有助于蠕虫病毒诱导的神经元凋亡。这些看似孤立的发现被合并为更广泛的模型，在本提案中描述。正如在各种非病毒凋亡模型中发现的那样，源自细胞膜的FAS信号传导可能通过通过FAS-DAXX-ass1-sek1-sek1-jnk凋亡途径进行细胞内JNK激活。我假设在病毒感染的背景下，DAXX是凋亡的特别重要的介体，因为其表达受宿主抗病毒免疫反应的控制。在AIM 1中，将利用主导阴性DAXX过表达（体内和体外）和siRNA介导的DAXX敲低模型（仅在体外）来确定DAXX在依伏病毒诱导的凋亡中的相对重要性。 AIM 2将采用免疫荧光成像和Western印迹技术来评估依孢病毒感染后DAXX的结合相互作用和信号传导作用。最后，AIM 3将解决DAXX表达受病毒感染宿主释放的干扰素（IFN）调节的可能性。总体而言，我假设IFN信号传导和相应的DAXX基因表达会使病毒感染的神经元容易受到凋亡。拟议的研究将探测病毒感染，免疫和细胞/组织/宿主死亡之间的界面，以加深我们对病毒宿主相互作用和神经元凋亡信号传导的理解。此外，这些研究将有可能建立理论基础，以最终开发神经保护药物。