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Mechanisms of gene regulation and RNA processing in synucleinopathies

突触核蛋白病中的基因调控和 RNA 加工机制

基本信息

批准号：
10447768
负责人：
Joseph R Mazzulli
金额：
$ 52.19万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10447768
关键词：
Axon Binding Brain Buffers Cell Death Cell Nucleus Cells Cellular Stress ChIP-seq Data Dementia Dementia with Lewy Bodies Detergents Disease Enzymes Event Exons Functional disorder Gene Expression Gene Expression Regulation Genes Genetic Genetic Transcription Health Human Inclusion Bodies Induced pluripotent stem cell derived neurons Knock-out Lead Lewy Bodies Lewy Body Dementia Link Liquid substance Measures Mediating Methods Midbrain structure Mitochondria Modeling Morphology Mutation Nerve Degeneration Neurites Neurodegenerative Disorders Neurons Nuclear Nuclear Proteins Nuclear RNA Parkinson Disease Parkinson&apos s Dementia Pathogenicity Pathologic Pathology Pathway interactions Patients Phase Phase Transition Phenotype Physiological Play Process Proteins Proteome Proteomics RNA RNA Editing RNA Processing RNA Splicing RNA-Binding Proteins RNA-specific adenosine deaminase 3 Regulation Role Seeds Solubility Stress Synapses Therapeutic Transcriptional Regulation Translations Untranslated RNA Virulence Factors Work age related neurodegeneration alpha synuclein axon guidance in vitro Model in vivo induced pluripotent stem cell knock-down mouse model neurotoxicity novel novel therapeutics prevent prion-like protein aggregation protein degradation protein transport proteostasis scaffold synucleinopathy trafficking transcription factor transcriptome sequencing

项目摘要

Neurodegenerative disorders including Lewy body Dementia (LBD) and Parkinson’s disease (PD) are characterized by aggregation of a-synuclein (a-syn), however the downstream toxic events that lead to cell death are not understood. Proteome dysfunction is a prominent feature of synucleinopathies, as indicated by genetics and pathology. To gain a comprehensive understanding of how the proteome changes in PD, we performed a quantitative proteomic study to identify proteins that aggregate in patient derived iPSC-neurons as a consequence of a-syn accumulation. By comparing iPSC neurons expressing A53T a-syn with isogenic corrected lines, we discovered a remarkable level of selectivity in the classes of proteins that aggregate. Specifically, we found that RNA binding proteins NONO and SFPQ undergo dramatic solubility shifts from detergent soluble into the insoluble state. NONO and SFPQ are multifunctional nuclear proteins that play critical roles in transcription regulation, RNA splicing, and RNA editing of genes that regulate axon guidance. They are core components of a membraneless sub-compartment in the nucleus called the paraspeckle, and contain prion- like low complexity domains that permit phase separation under physiological conditions. Paraspeckles occur in neuronal cultures and in vivo in the brain, and are thought to play key roles in regulating homeostatic stress by transiently sequestering transcription factors and RNAs to prevent translation. Once stress subsides, paraspeckles normally dissolve and gene expression returns to normal. However, we have found that NONO and SFPQ irreversibly form pathological aggregates in patient iPSC-neurons and LBD patient brain. This effect is specifically associated with a-syn accumulation, and does not occur with general cellular stress. Mechanistic studies in iPSC-neurons suggest that formation of NONO/SFPQ aggregates is associated with loss of their functions, resulting in neurite degeneration. We find that the SFPQ transcriptional target, ADAR3 that mediates RNA editing, is nearly completely depleted in patient neurons. Here, we propose to examine the mechanism of how a-syn accumulation leads to aberrant NONO/SFPQ aggregation in the nucleus and downstream pathophysiology. Given their role in RNA splicing and editing, we propose to employ both targeted and unbiased methods to identify changes in RNA editing including RNA-seq, exon-junction microarrays to examine RNA splicing, and ChIP-seq to detect changes in SFPQ transcriptional activity. These phenotypes will be correlated with distinct aggregated forms of a-syn and neurodegeneration. Finally, we will attempt to rescue established phenotypes in patient iPSC-neurons by promoting soluble, function NONO/SFPQ. Our preliminary studies have identified a novel pathogenic pathway in synucleinopathies, and we will extend these findings by examining the mechanisms of gene dysregulation and RNA processing. We will provide the first description of RNA editing and splicing changes in patient iPSCs, which may uncover novel disease mechanisms and therapeutic strategies.

神经退行性疾病包括路易体痴呆（LBD）和帕金森病（PD）