权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Investigating RNA dysregulation in Neurological Disease through study of Pontocerebellar Hypoplasia Type 1b

通过 1b 型桥小脑发育不全研究来调查神经系统疾病中的 RNA 失调

基本信息

批准号：
10638196
负责人：
Derrick Morton
金额：
$ 42.84万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
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：
2023-04-15 至 2028-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10638196
关键词：
Affect Alleles Amino Acid Substitution Amino Acids Atrophic Attention Behavior Biochemical Biological Biological Models Biology Biophysics Brain Brain region Breathing CCRL2 gene CRISPR/Cas technology Catalytic Domain Cell Nucleus Cell model Cells Cerebellum Child Childhood Clinical Complement Complex Cytoplasm Data Defect Development Developmental Delay Disorders Disease Drosophila genus Enhancers Etiology Fishes Gene Expression Gene Expression Regulation Genes Genetic Genetic Screening Genomics Genotype Goals Heterogeneity Homeostasis Homologous Gene Human Impairment In Situ Individual Inherited Life Link Longevity Mass Spectrum Analysis Methods Microcephaly Missense Mutation Modeling Molecular Molecular Machines Motor Movement Muscular Atrophy Mutation Nervous System Neurodevelopmental Disorder Neuronal Dysfunction Neurons Pancreatic ribonuclease Pathology Phenotype Play Pontine structure Pontocerebellar hypoplasia Post-Transcriptional RNA Processing Post-Transcriptional Regulation RNA RNA Decay RNA Processing Reporting Ribonucleases Role Series Severities Spinal Cord System Technology Testing Tissues Transcript Variant Work autosome behavioral phenotyping brain morphology dosage exosome fly in vivo in vivo Model insight motor behavior motor learning mutant nervous system disorder neurodevelopment novel palliative sensory system single-cell RNA sequencing tool transcriptome transcriptome sequencing transcriptomics

项目摘要

PROJECT SUMMARY/ABSTRACT Pontocerebellar Hypoplasia Type 1b (PCH1b) is an autosomal recessive neurological disorder characterized by hypoplasia/atrophy of the cerebellum and pons that is often fatal within the first year of life. The cerebellum and pons integrate information from sensory systems, the spinal cord, and other parts of the brain to regulate motor movements, breathing, and learning motor behavior. Individuals with PCH1b show muscle atrophy/weakness, microcephaly, and developmental delay. Most individuals with PCH1b do not live past childhood and current treatment is purely palliative. Mutations that cause PCH1b occur in the EXOSC3 gene, which encodes a structural cap subunit of an evolutionarily conserved and ubiquitously expressed RNA processing complex, the RNA exosome. The RNA exosome is a ribonuclease composed of both structural and catalytic subunits that play a critical role in the post-transcriptional regulation of RNA. This complex is required for 3’ to 5’ processing and degradation of a vast number of RNAs in both the nucleus and cytoplasm. Post-transcriptional processing of RNA is a critical regulatory step in gene expression, as underscored by the number of neurological diseases caused by defects in RNA processing factors. The tissue-specific phenotypes caused by the RNA exosome complex are challenging to understand based on current models of RNA exosome function with only limited analysis of the complex in any multicellular model in vivo. Thus, we aim to investigate the in vivo functional consequences of distinct disease-causing amino acid substitutions in EXOSC3 that are linked to a range of mild to severe phenotypes in PCH1b. We have generated an allelic series of EXOSC3 disease-linked missense mutations in the Drosophila orthologue Rrp40 via CRISPR/Cas9 editing technology. Our previous work in flies revealed an enhanced requirement for Rrp40 in neurons. Furthermore, our RNA-seq analysis of brain-enriched transcriptomes of Rrp40 mutants revealed increases in steady-state levels of functionally important neuronal transcripts, suggesting that disease-causing amino acid changes in the Drosophila RNA exosome subunit Rrp40 contribute to neuronal dysfunction. Our goal now is to characterize how disease-causing amino acid substitutions in Rrp40 alter the molecular and cellular landscape of the developing nervous system in Drosophila in vivo. We will test the hypothesis that the RNA exosome regulates RNAs that are critical for proper neurodevelopment and function, a distinct subset of which are regulated by the RNA exosome cap subunit Rrp40 (EXOSC3) through three complementary aims: 1) Assess the functional consequences of amino acid substitutions in the Drosophila RNA exosome subunit Rrp40 corresponding to those that cause PCH1b; 2) Interrogate how Rrp40 mutations affect expression and localization of key neuronal transcripts within the fly brain; and 3) Exploit a genetic screen to determine whether aberrant accumulation of specific RNA exosome targets disrupt fly development and/or homeostasis. Successful completion on these aims will provide a synergistic understanding of RNA exosome biology and invaluable insights into PCH1b disease etiology.

项目总结/摘要脑桥小脑发育不全1b型（PCH 1b）是一种常染色体隐性遗传神经系统疾病，其特征在于：小脑和脑桥的发育不全/萎缩，通常在生命的第一年内致命。小脑和脑桥整合来自感觉系统、脊髓和大脑其他部分的信息来调节运动运动、呼吸和学习运动行为。患有PCH 1b的个体表现出肌肉萎缩/无力，小头畸形和发育迟缓大多数患有PCH 1b的人没有活过童年，治疗纯粹是治标不治本。导致PCH 1b的突变发生在EXOSC 3基因中，该基因编码一种结构帽亚基的进化保守和普遍表达的RNA加工复合物， RNA外泌体。RNA外泌体是由结构和催化亚基组成的核糖核酸酶，在RNA的转录后调控中起关键作用。这种复合物是3'至5'加工所需的，细胞核和细胞质中大量RNA的降解。的转录后加工 RNA是基因表达的关键调节步骤，正如神经系统疾病的数量所强调的那样。是由RNA加工因子缺陷引起的。RNA外泌体引起的组织特异性表型基于目前的RNA外泌体功能模型，在体内任何多细胞模型中分析复合物。因此，我们的目的是研究在体内的功能， EXOSC 3中不同的致病氨基酸取代的后果，这些取代与一系列轻度的 PCH 1b的严重表型。我们已经产生了一个等位基因系列的EXOSC 3疾病相关的错义通过CRISPR/Cas9编辑技术在果蝇直向同源物Rrp 40中的突变。我们以前在苍蝇方面的工作揭示了神经元对Rrp 40的需求增强。此外，我们对大脑富集的 Rrp 40突变体的转录组显示功能重要的神经元的稳态水平增加，转录，表明果蝇RNA外泌体亚基Rrp 40中的致病氨基酸变化导致神经元功能障碍我们现在的目标是描述致病氨基酸替换在Rrp 40中的表达改变了果蝇体内发育神经系统的分子和细胞景观。我们将测试RNA外泌体调节RNA的假设，RNA对正常的细胞生长至关重要。神经发育和功能，其中一个独特的子集由RNA外泌体帽调控亚基Rrp 40（EXOSC 3）通过三个互补的目的：1）评估氨基的功能后果果蝇RNA外泌体亚基Rrp 40中对应于导致PCH 1b的酸取代; 2）探究Rrp 40突变如何影响果蝇内关键神经元转录物的表达和定位脑;和3）利用遗传筛选来确定是否特异性RNA外泌体的异常积累靶扰乱果蝇发育和/或体内平衡。成功实现这些目标将为 RNA外泌体生物学的协同理解和对PCH 1b疾病病因学的宝贵见解。