Effects of TDP-43 Pathology on Innate Antiviral Mechanisms in Neurodegenerative Disease

TDP-43 病理学对神经退行性疾病先天抗病毒机制的影响

• 批准号: 10537727
• 负责人: Stephanie Jackvony
• 金额: $ 4.68万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10537727
• 关键词: Affect; Alternative Splicing; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Antiviral Response; Astrocytes; Binding; Brain; Cells; Cytoplasm; DNA Transposable Elements; DNA-Binding Proteins; Data; Dementia; Disease; Disease Progression; Exposure to; Frontotemporal Dementia; Functional disorder; Gene Expression; Genes; Genetic; Genetic Transcription; Herpesvirus 1; Hippocampus (Brain); Human; Immune; Immune response; Impairment; In Vitro; Infection; Interferons; Laboratories; Link; Molecular; Monitor; Mus; Natural Immunity; Nerve Degeneration; Neurodegenerative Disorders; Neuroimmune; Neurologic; Neurons; Nuclear; Onset of illness; Outcome Study; Pathogenicity; Pathologic; Pathology; Pathway interactions; Pharmacology; Physiological; Plaque Assay; Poly C; Poly I-C; Predisposition; Process; Proteins; RNA; RNA Binding; RNA Processing; RNA Recognition Motif; RNA Splicing; RNA Transport; RNA-Binding Proteins; Signal Transduction; Specific qualifier value; Techniques; Testing; Transcriptional Regulation; Variant; Viral; Virus Diseases; Western Blotting; adeno-associated viral vector; antimicrobial; base; case control; cell type; differential expression; effective therapy; in vivo; in vivo Model; interest; knock-down; mouse model; mutant; new therapeutic target; non-demented; novel; overexpression; pathogen; pathogenic virus; prevent; protein TDP-43; relating to nervous system; response; risk variant; single-cell RNA sequencing; therapeutic target

Project Summary/Abstract Dysregulation of transactivating response region DNA-binding protein-43 (TDP-43) has been linked to many neurodegenerative diseases, including frontotemporal dementia, amyotrophic lateral sclerosis, and Alzheimer’s disease (AD). TDP-43 has a variety of functions linked to its RNA-binding motif, including regulation of transcription, splicing, and RNA transport. Along with these effects, TDP-43 alters expression of interferon (IFN)- related and other immune genes essential for antiviral responses. The relationship between viral pathogens and TDP-43 is bidirectional, as exposure to poly(I:C), which simulates viral pathogens, can promote subcellular mislocalization of TDP-43. Viral pathogens, like TDP-43 dysregulation, are linked to AD and other dementias; AD has been associated with increased presence of viral pathogens, like herpes simplex virus 1 (HSV-1), and altered IFN-related signaling and neuroimmune cascades. Our laboratory found that, like neuronal TDP-43, astrocytic TDP-43 can be mislocalized to the cytoplasm in AD. Dysregulation of astrocytic TDP-43 in mouse models caused neural deficits and cell-autonomous changes in antiviral and IFN-inducible factors. Further, dysregulated TDP-43 increased astrocytic susceptibility to HSV-1. Astrocytic susceptibility to HSV-1 associated with overexpression of human TDP-43 was reduced by blocking the ability of human TDP-43 to bind RNA. Previous studies also show that the RNA-binding domain on TDP-43 is necessary for its other disease-linked effects. Based on this evidence, I will test the hypothesis that dementia-related TDP-43 dysfunction affects antiviral pathways and increases neural susceptibility to HSV-1 by altering TDP-43 binding to host RNA, resulting in aberrant host antiviral and immune gene expression and impaired innate antiviral signaling. I propose to use a variety of cellular and molecular techniques to examine in vitro (Aim 1.1) and in vivo (Aim 1.2) susceptibility to HSV-1 following cell-specific expression of TDP-43 variants that either maintain nuclear localization, mislocalize to the cytoplasm, cannot bind to RNA, or both. I will also determine cell-specific molecular mechanisms that promote differences in antiviral pathways via single-cell RNA sequencing (Aim 2.1), and conduct targeted analysis of alternative splicing (ScISOr-Seq), transposable element expression (TEtranscripts), and protein levels (Western blotting). Finally, I will examine the physiological functions of differential genes of interest identified in Aim 2.1 using genetic and pharmacological approaches. Uncovering the mechanistic links that connect TDP-43 dysregulation to antiviral pathways and viral susceptibility may define new pathobiological mechanisms and therapeutic targets to prevent neurodegenerative disease onset and progression.

项目概要/摘要 反式激活反应区 DNA 结合蛋白 43 (TDP-43) 的失调与许多疾病有关 神经退行性疾病，包括额颞叶痴呆、肌萎缩侧索硬化症和阿尔茨海默病 疾病（AD）。 TDP-43 具有与其 RNA 结合基序相关的多种功能，包括调节 转录、剪接和 RNA 运输。除了这些作用之外，TDP-43 还改变干扰素 (IFN) 的表达 - 相关基因和其他抗病毒反应所必需的免疫基因。病毒与病原体的关系 TDP-43 是双向的，因为暴露于模拟病毒病原体的聚 (I:C) 可以促进亚细胞 TDP-43 的错误定位。病毒病原体，如 TDP-43 失调，与 AD 和其他痴呆症有关。 AD 与病毒病原体的增加有关，例如单纯疱疹病毒 1 (HSV-1)，并且 改变 IFN 相关信号传导和神经免疫级联反应。我们的实验室发现，与神经元 TDP-43 一样， AD 中星形细胞 TDP-43 可能错误定位至细胞质。小鼠星形胶质细胞 TDP-43 失调 模型引起神经缺陷以及抗病毒和干扰素诱导因子的细胞自主变化。更远， TDP-43 失调会增加星形胶质细胞对 HSV-1 的易感性。星形胶质细胞对 HSV-1 的易感性相关 通过阻断人 TDP-43 结合 RNA 的能力，可以减少人 TDP-43 的过度表达。 先前的研究还表明，TDP-43 上的 RNA 结合结构域对于其他与疾病相关的疾病是必需的。 影响。基于这一证据，我将检验以下假设：痴呆相关的 TDP-43 功能障碍会影响 通过改变 TDP-43 与宿主 RNA 的结合，从而增加抗病毒途径并增加对 HSV-1 的神经敏感性，从而 宿主抗病毒和免疫基因表达异常以及先天抗病毒信号传导受损。我建议使用 各种细胞和分子技术来检查体外（目标 1.1）和体内（目标 1.2）的敏感性 TDP-43 变体细胞特异性表达后的 HSV-1 要么维持核定位，要么错误定位 结合到细胞质，不能结合 RNA，或两者兼而有之。我还将确定细胞特异性的分子机制 通过单细胞RNA测序促进抗病毒途径的差异（目标2.1），并进行有针对性的研究 选择性剪接 (ScISOr-Seq)、转座元件表达 (TEtranscripts) 和蛋白质分析 水平（蛋白质印迹）。最后，我将检查感兴趣的差异基因的生理功能 使用遗传和药理学方法在目标 2.1 中确定。揭示其中的机械联系 将 TDP-43 失调与抗病毒途径和病毒易感性联系起来可能会定义新的病理生物学 预防神经退行性疾病发生和进展的机制和治疗靶点。