Determine the neurotoxicity of RNA metabolism dysfunction caused by cytoplasmic TDP-43 aggregates

确定细胞质 TDP-43 聚集体引起的 RNA 代谢功能障碍的神经毒性

基本信息

批准号：
10730167
负责人：
Sarah H Shahmoradian
金额：
$ 61.71万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-19 至 2028-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10730167
关键词：
ALS patients Affect Alzheimer&apos s Disease Alzheimer&apos s disease patient Alzheimer&apos s disease related dementia Architecture Autopsy Brain Cell membrane Cells Complex Core Assembly Cytoplasm Cytoplasmic Granules Dementia Disease Elderly Electron Microscopy Frontotemporal Dementia Functional disorder Half-Life Hippocampus Image Immediate-Early Genes Immunohistochemistry Impairment Liquid substance Membrane Messenger RNA Morphology Motor Neurons Mus Mutation Names Neurodegenerative Disorders Neurons Nonsense-Mediated Decay Organelles Pathologic Pathology Patients Phase Phase Transition Physical condensation Post-Translational Protein Processing Process Proteins Proteomics RNA RNA Processing RNA metabolism RNA-Binding Proteins Reporting Ribonucleoproteins Sampling Spinal Cord Stress Syndrome TDP-43 aggregation Testing Tissues Toxic effect Translational Repression age related in vivo induced pluripotent stem cell innovation light microscopy limbic-predominant age-related TDP-43 encephalopathy mRNA Decay mRNA Transcript Degradation neuronal cell body neuropathology neurotoxicity prion-like protein TDP-43 protein aggregation protein complex response

项目摘要

This proposal seeks to determine how TDP-43 protein aggregates dysregulate P-body function in neurons and subsequently produce neurotoxicity. Cytoplasmic aggregation of TDP-43 has been reported in nearly every age-dependent neurodegenerative disease, including in >40% of frontotemporal dementia (FTD), in the hippocampal neurons of Alzheimer’s disease (AD) patients, in >90% of ALS. It also defines a recently recognized AD-like dementia in the oldest elderly, an AD-like syndrome named Limbic-predominant Age- related TDP-43 Encephalopathy (LATE). We have identified that TDP-43 cytoplasmic aggregates regulate the liquid-liquid phase separation (LLPS) of RNA processing bodies (P-bodies) in neuron-like cells and postmortem spinal cord motor neurons in ALS patients. P-bodies are cytoplasmic membraneless ribonucleoprotein (RNP) granules composed of RNAs and protein complexes involved in translational repression and mRNA decay. Neurons carry a high number of P-bodies in the soma. We hypothesize that TDP-43 aggregation causes neuronal toxicity by disrupting the morphology and function of P-bodies. Our proposal is highly innovative because how TDP-43 proteinopathy regulates the LLPS of other membraneless organelles has not been reported in vivo. We propose to use cutting-edge imaging, proteomic, and sequencing approaches to determine the protein and RNA composition of neuronal P-bodies and how it changes in response to TDP-43 aggregation in vivo. We will first determine how TDP-43 cytoplasmic aggregates initiate P-body disassembly and then determine the RNA metabolism change in neurons carrying TDP-43 aggregates or defective P-bodies. Lastly and importantly, we will determine whether P-body proteins and RNA can serve as pathological markers for AD-related dementia, such as LATE.

该提案旨在确定TDP-43蛋白聚集体如何使P体功能失调神经元并随后产生神经毒性。 TDP-43的细胞质聚集已报道几乎所有依赖年龄的神经退行性疾病，包括额叶痴呆症的40％（FTD），在> 90％的ALS中，在阿尔茨海默氏病（AD）患者的海马神经元中。它也定义了最近在古老的老古老的广告状痴呆症中被公认的一种广告综合症，名为Libbic-Predominatimonal-age- 相关的TDP-43脑病（晚期）。我们已经确定TDP-43细胞质聚集体调节神经元样细胞中的RNA加工体（P-Bodies）的液态液相分离（LLP）和 ALS患者的后脊髓运动神经元。 p-bodies是细胞质膜无核糖核蛋白（RNP）颗粒由RNA和蛋白质复合物组成，参与转化表达和mRNA衰变。神经元在躯体中含有大量的p-bodies。我们假设TDP-43 聚集通过破坏p体的形态和功能来引起神经元毒性。我们的提案具有很高的创新性，因为TDP-43蛋白质病如何调节其他无膜的LLP 细胞器尚未在体内报告。我们建议使用最先进的成像，蛋白质组学和测序确定神经元P-体的蛋白质和RNA组成的方法，以及它在反应中的变化到TDP-43在体内聚集。我们将首先确定TDP-43细胞质聚集体如何启动P体拆卸，然后确定携带TDP-43骨料的神经元的RNA代谢变化或 p身体有缺陷。最后，重要的是，我们将确定P体蛋白和RNA是否可以用作广告相关痴呆的病理标记，例如迟到。