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: 10689062
• 负责人: Stephanie Jackvony
• 金额: $ 4.77万
• 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/10689062
• 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; Herpes Simplex Infections; Herpesvirus 1; Hippocampus; Human; Immune; Immune response; Impairment; In Vitro; Injections; Interferons; Laboratory Finding; Link; Molecular; Monitor; Mus; Natural Immunity; Nerve Degeneration; Neurodegenerative Disorders; Neuroimmune; Neurologic; Neurons; Nuclear; Onset of illness; Outcome; Pathogenicity; Pathologic; Pathology; Pathway interactions; Physiological; Plaque Assay; 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; case control; cell type; differential expression; effective therapy; in vivo; in vivo Model; interest; knock-down; mouse model; mutant; neural; new therapeutic target; non-demented; novel; overexpression; pathogen; pathogenic virus; pharmacologic; prevent; protein TDP-43; response; risk variant; single-cell RNA sequencing; therapeutic target; transcriptome sequencing

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.

项目摘要/摘要 反式激活反应区DNA结合蛋白-43(TDP-43)的失调与许多 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氏症 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。除了这些作用外，TDP-43还改变了干扰素的表达- 抗病毒反应所必需的相关和其他免疫基因。病毒病原体与人类免疫功能的关系 TDP-43是双向的，因为接触模拟病毒病原体的聚(I：C)可以促进亚细胞 TDP-43基因定位错误。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，如单纯疱疹病毒1(HSV-1)，以及 改变干扰素相关信号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， 星形细胞TDP-43在AD中可错误定位于细胞质。小鼠星形胶质细胞TDP-43的表达异常 模型导致抗病毒和干扰素诱导因子的神经缺陷和细胞自主性改变。此外， TDP-43表达异常增加了星形细胞对HSV-1的易感性。星形细胞对HSV-1的易感性 人TDP-43的过表达可通过阻断人TDP-43与RNA的结合能力而降低。 以往的研究也表明，TDP-43上的RNA结合域对于其他与疾病相关的疾病是必要的 效果。基于这一证据，我将检验与痴呆症相关的TDP-43功能障碍影响 通过改变TDP-43与宿主RNA的结合，抗病毒途径和增加神经对HSV-1的敏感性，从而导致 在异常宿主中，抗病毒和免疫基因表达和先天抗病毒信号受损。我建议使用 多种细胞和分子技术检测体外(AIM 1.1)和体内(AIM 1.2)对 HSV-1跟随TDP-43变异体的细胞特异性表达，这些变异体要么维持核定位，要么错位定位 到细胞质，不能与RNA结合，或两者兼而有之。我还将确定特定于细胞的分子机制 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 选择性剪接(ScISOr-Seq)、转座元件表达(TE转录本)和蛋白质分析 表达水平(Western Blotting)。最后，我将研究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。揭示了机械联系 将TDP-43异常与抗病毒途径和病毒易感性联系起来可能定义新的病理生物学 预防神经退行性疾病发生和发展的机制和治疗靶点。