Aberrant P-bodies accumulation and clearance in yeast and human cells.

酵母和人体细胞中异常 P 体的积累和清除。

基本信息

批准号：
10390634
负责人：
Nava Segev
金额：
$ 23.99万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
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：
2022-01-18 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10390634
关键词：
Address Adverse effects Affect Autophagocytosis Behavior Biological Biological Models Brain Caring Cell Line Cell Survival Cell model Cell physiology Cells Child Cytoplasmic Granules Data Defect Development Disease Economic Burden Enzymes Etiology Family Fibroblasts Genes Goals Growth Human Individual Intellectual functioning disability Knowledge Lead Liquid substance Lysosomes Membrane Messenger RNA Missense Mutation Mission Modeling Molecular Genetics Mutation National Institute of Neurological Disorders and Stroke Neurites Neurodevelopmental Disorder Neurons Nonsense-Mediated Decay Organelles Pathogenicity Pathway interactions Patients Phase Phenotype Play Population Predisposition Proteins RNA RNA Processing RNA helicase A Regulation Research Role Severities Societies Stress Synaptic plasticity System Toxic effect Transfection Ubiquitin Variant Yeasts base cell growth cell immortalization cell motility de novo mutation early onset genome sequencing helicase human disease human tissue insight mRNA Decay multicatalytic endopeptidase complex mutant novel novel therapeutic intervention response severe intellectual disability tissue/cell culture yeast protein

项目摘要

Abstract RNA-protein (RNP) granules are dynamic membrane-less organelles that form during normal growth and in response to stress in a reversible manner. One type of RNP granules is Processing Bodies (P-bodies, or PBs), which contain mRNA together with RNA processing enzymes. Their roles include sequestration of malfunctioning mRNA destined for degradation and storing and silencing of mRNAs when not needed. PBs components and behavior are highly conserved between yeast and human cells. Our understanding of composition and assembly by liquid-phase separation normal PBs has progressed in the last decade. In contrast, our knowledge about involvement of PBs in human disease is scarce as is evidence about the occurrence of aberrant PBs and clearance of normal or aberrant PBS. In this project, we propose to study variants in a PBs component associated with a neurodevelopmental disorder that causes intellectual disability. Currently, there is no cure for such disorders. We hypothesize that the variants cause accumulation of aberrant PBs that, unlike normal PBs, are not reversible. The human variants carry missense mutations in residues identical in the yeast protein. Therefore, we started by modeling the effects of these mutations on RNA processing and accumulation of PBs in yeast. While normal PBs assemble and disassemble according to cellular needs, our preliminary evidence in yeast cells points to accumulation of persistent PBs as the major phenotype of these mutations. We propose to study the effects of these mutations in yeast and in human tissue culture cells, including neuronal cell lines. The phenotypes we propose to assess are on the accumulation and dynamics of persistent PBs, including adverse effects they might have on cell function. In addition, we will explore possible pathways that can clear normal and persistent PBs, such as macro- and micro-autophagy and ubiquitin-associated degradation. For these studies we will use a combination of molecular genetics and cellular approaches. Most approaches for studying RNP granules, granule clearance pathways in yeast and human cells are established in our lab. Achieving the goals of this proposal would provide novel paradigms on existence and behavior of persistent PBs and the role they play in a neurodevelopmental disorder. Moreover, identifying pathways that can clear normal and aberrant PBs would provide novel therapeutic strategies for a neurodevelopmental disorder associated with intellectual disability, in line with the missions of the National Institute of Neurological Disorders and Stroke.

抽象的 RNA 蛋白 (RNP) 颗粒是在正常情况下形成的动态无膜细胞器。生长和以可逆的方式应对压力。一种类型的 RNP 颗粒是加工体（P-体，或 PB），包含 mRNA 和 RNA 加工酶。它们的作用包括隔离注定要降解的故障 mRNA 不需要时存储和沉默 mRNA。 PB 组件和行为是在酵母和人类细胞之间高度保守。我们对构图的理解和通过液相分离普通PB的组装在过去十年中取得了进展。相比之下，我们对 PB 与人类疾病的关系知之甚少，关于 PB 参与人类疾病的证据也是如此。异常 PB 的发生以及正常或异常 PBS 的清除。在这个项目中，我们建议研究与导致智力障碍的神经发育障碍。目前尚无治愈方法此类疾病。我们假设这些变异会导致异常 PB 的积累，与普通 PB 不同，它们是不可逆的。人类变异体在残基中携带错义突变与酵母蛋白相同。因此，我们首先对这些突变的影响进行建模酵母中 RNA 加工和 PB 积累的影响。虽然普通 PB 会组装并根据细胞需要进行拆卸，我们在酵母细胞中的初步证据表明持久性 PB 的积累是这些突变的主要表型。我们建议研究这些突变对酵母和人体组织培养细胞的影响，包括神经元细胞系。我们建议评估的表型是积累和持久性 PB 的动态，包括它们可能对细胞功能产生的不利影响。在此外，我们将探索清除正常和持续性PB的可能途径，例如宏观和微观自噬以及泛素相关降解。对于这些研究，我们将结合使用分子遗传学和细胞方法。大多数学习方法我们的实验室建立了 RNP 颗粒、酵母和人体细胞中的颗粒清除途径。实现该提案的目标将为存在和行为提供新颖的范式持续性 PB 及其在神经发育障碍中的作用。此外，识别清除正常和异常 PB 的途径将为以下疾病提供新的治疗策略：与智力障碍相关的神经发育障碍，符合国家神经疾病和中风研究所。