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 蛋白聚集体如何失调 P 体功能神经元并随后产生 TDP-43 的细胞质聚集。几乎所有与年龄相关的神经退行性疾病，包括 >40% 的额颞叶痴呆 (FTD)，在阿尔茨海默氏病 (AD) 患者的海马神经元中，> 90% 的 ALS 中也存在这种现象。在最年长的老年人中发现了类似 AD 的痴呆症，一种名为 Limbic-predomined Age- 的类似 AD 的综合征相关的 TDP-43 脑病 (LATE) 我们已经确定 TDP-43 细胞质聚集体调节神经元样细胞中 RNA 加工体（P 体）的液-液相分离 (LLPS) 和 ALS 患者死后的脊髓运动神经元是细胞质无膜的。由参与翻译抑制的 RNA 和蛋白质复合物组成的核糖核蛋白 (RNP) 颗粒神经元在体细胞中携带大量 P 体，我们与 TDP-43 进行了斗争。聚集通过破坏 P 体的形态和功能而引起神经元毒性。该提案具有高度创新性，因为 TDP-43 蛋白病如何调节其他无膜细胞的 LLPS 我们建议使用尖端成像、蛋白质组学和测序技术。确定神经元 P 体的蛋白质和 RNA 组成及其响应变化的方法我们将首先确定 TDP-43 细胞质聚集体如何启动 P-body。分解，然后确定携带 TDP-43 聚集体的神经元中 RNA 代谢的变化或最后也是重要的是，我们将确定 P 体蛋白和 RNA 是否可以充当缺陷的 P 体。 AD 相关痴呆的病理标志物，例如 LATE。