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：
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
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/10537727
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 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

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.

项目摘要/摘要反式激活反应区域DNA结合蛋白-43（TDP-43）的失调与许多神经退行性疾病，包括额颞痴呆，肌萎缩性侧面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传输。与这些效果一起，TDP-43改变了干扰素的表达（IFN） - 相关和其他免疫基因对于抗病毒反应必不可少。病毒病原体与 TDP-43是双向的，因为暴露于模拟病毒病原体的聚（I：C）可以促进亚细胞亚细胞 TDP-43的错误定位。病毒病原体，例如TDP-43失调，与AD和其他痴呆症有关。 AD与病毒病原体的存在增加有关，例如单纯疱疹病毒1（HSV-1）和 IFN相关的信号传导和神经免疫性级联反应改变。我们的实验室发现，像神经元TDP-43一样星形胶质细胞TDP-43可能被错误地定位于AD中的细胞质。小鼠星形细胞TDP-43的失调模型导致神经元定义了抗病毒和IFN诱导因素的细胞自主变化。更远， TDP-43失调增加了星形胶质细胞对HSV-1的敏感性。星形胶质细胞对HSV-1相关的敏感性通过阻止人TDP-43结合RNA的能力，可以减少人类TDP-43的过表达。先前的研究还表明，TDP-43上的RNA结合域对于其他疾病连接是必需的效果。基于此证据，我将检验以下假设：痴呆相关的TDP-43功能障碍会影响抗病毒途径并通过改变与宿主RNA的TDP-43结合来增加对HSV-1的神经敏感性，从而导致在异常的宿主抗病毒和免疫表达以及造成的先天抗病毒信号传导中。我建议使用多种细胞和分子技术以检查体外（AIM 1.1）和体内（AIM 1.2）敏感性 TDP-43变体的细胞特异性表达后的HSV-1，要么维持核定位，要么置置不稳定对于细胞质，不能与RNA或两者结合。我还将确定细胞特异性的分子机制通过单细胞RNA测序（AIM 2.1）促进抗病毒途径的差异，并进行针对性替代剪接（Scisor-Seq），转座元元素表达（Tetranscripts）和蛋白质的分析水平（蛋白质印迹）。最后，我将检查感兴趣的差异基因的物理功能使用遗传和药物方法在AIM 2.1中识别。发现机械链接将TDP-43失调连接到抗病毒途径，病毒易感性可能定义新的病理学预防神经退行性疾病发作和进展的机制和治疗靶标。