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

基本信息

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
项目类别：
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&apos 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

项目摘要

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 TDP-43 具有与其 RNA 结合基序相关的多种功能，包括调节除了这些作用外，TDP-43 还改变了干扰素 (IFN) 的表达。病毒病原体和抗病毒反应所必需的相关和其他免疫基因之间的关系。 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，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，小鼠中星形细胞 TDP-43 失调。模型引起抗病毒和干扰素诱导因子的神经缺陷和细胞自主变化。 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 相关的易感性。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 结合域对于其其他与疾病相关的疾病是必需的。基于这一证据，我将检验痴呆相关 TDP-43 功能障碍影响的假设。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，从而在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的情况下，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，或者两者兼而有之，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) 和蛋白质分析最后，我将检查感兴趣的差异基因的生理功能。目标 2.1 中使用遗传和药理学方法发现了其中的机制联系。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。