PRC1-dependent H2A Monoubiquitination During Human Corticogenesis

人类皮质生成过程中 PRC1 依赖性 H2A 单泛素化

基本信息

批准号：
10604561
负责人：
Charles William Ryan
金额：
$ 4.05万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
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：
2023-01-01 至 2025-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10604561
关键词：
3-Dimensional Accounting Affect Alleles Apoptosis Apoptotic BMI1 gene Binding Binding Sites Brain Catalysis Cell Cycle Cerebrum Child Chromatin Compensation Complement Complex DNA Damage DNA Repair DNA Sequence Alteration DNA replication fork Data Databases Defect Deposition Developed Countries Development Disease Dominant-Negative Mutation Double Strand Break Repair Environment Future Gene Expression Gene Silencing Genes Genetic Genetic Transcription Goals Growth Histone H2A Histones Human Individual Infant Intellectual functioning disability Ionizing radiation Knock-out Knowledge Lead Learning Light Literature Lymphocyte Mediating Medical Genetics Microcephaly Mitotic Modeling Molecular Monoubiquitination Mutation Neurodevelopmental Disorder Neurons Nucleosomes Organoids PRC1 Protein Pathogenesis Pathogenicity Patients Physicians Population Post-Translational Protein Processing Process Proliferating Prosencephalon Proteins Reporting Resolution Response Elements Role Schizophrenia Scientist Site Testing Therapeutic Training Transcriptional Regulation Ubiquitination Variant brain size career catalyst cell type clinically relevant crosslink developmental disease early onset experimental study functional restoration gene repression genetic testing genome integrity genome sequencing genome-wide human embryonic stem cell human embryonic stem cell line insight mouse model mutant nerve stem cell neural novel postmitotic premature prevent rapid growth repaired targeted treatment trend ubiquitin-protein ligase whole genome

项目摘要

PROJECT SUMMARY Intellectual disability affects 2-3% of the population in industrialized nations, with genetic causes accounting for approximately half of these cases. Whole genome sequencing has led to the discovery of several novel pathogenic variants underlying neurodevelopmental disorders. Among these variants, proteins involved in chromatin post-translational modifications are highly represented. Variants in the polycomb repressive complex 1 (PRC1), the primary catalyst of histone 2A monoubiquitination (H2AUb1), illustrate this trend. RING1 and RNF2 are paralogues that serve as the constituent E3 ubiquitin ligase of PRC1, and missense variants in these genes that blunt H2AUb1 catalysis are associated with a neurodevelopmental disorder characterized by microcephaly, intellectual disability, and early onset schizophrenia. The goal of this proposal is to understand how pathogenic RING1 variants lead to disease genetically and molecularly. To explore the role of PRC1- dependent H2AUb1 in the context of human corticogenesis, we have generated isogenic human embryonic stem cell (hESC) lines harboring pathogenic variants in RING1 and subjected them to neural differentiation to generate 3D cerebral organoids. Previous studies suggest that PRC1-dependent H2AUb1 has roles in transcriptional repression and double strand break repair. Consistent with these previous reports, our preliminary data demonstrates that the RING1G284A/G284A variant diminishes global H2AUb1 levels, leads to increased expression of pro-apoptotic genes and decreased growth in cerebral organoids, and disrupts double strand break repair in neural progenitor cells (NPCs). Aim 1 will explore the genetic mechanism of RING1 pathogenic variants, as experiments will shed light onto whether pathogenesis is due to lost activity of wild type RING1 or due to a dominant negative effect of the mutant allele. Aim 2 will evaluate the molecular mechanism by which pathogenic RING1 variants cause microcephaly. Experiments will evaluate NPCs and organoids differentiated from hESCs harboring RING1 pathogenic variants for defects in DNA damage repair, increased rates of apoptosis, and the ability to silence transcription at sites of DNA damage. Mechanistic insights learned from these experiments will shed light onto the role of PRC1-dependent H2AUb1 in proliferating NPCs and in maintaining genome integrity more broadly. In addition, the results of these studies will inform future efforts to devise therapeutic strategies to restore these functions in individuals with pathogenic RING1 variants.

项目摘要智力残疾影响工业化国家的2-3％的人口，遗传原因是这些案件中约有一半。整个基因组测序导致发现了几本小说神经发育障碍的基础变种。在这些变体中，涉及的蛋白质染色质后翻译修饰是高度表示的。 Polycomb抑制剂复合物中的变体 1（PRC1），组蛋白2a单泛素化（H2AUB1）的主要催化剂，说明了这一趋势。 ring1和 RNF2是PRC1的组成E3泛素连接酶的旁产钝H2AUB1催化的基因与以神经发育障碍有关小头畸形，智力残疾和早期发作精神分裂症。该提议的目的是了解致病性环1变体如何导致遗传和分子疾病。探索Prc1-的作用依赖性的H2AUB1在人皮质生成的背景下，我们产生了等源性人类胚胎干细胞（hESC）线，在RING1中具有致病性变异，并将其进行神经分化至产生3D大脑器官。先前的研究表明，依赖PRC1的H2AUB1在转录抑制和双链断裂修复。与以前的报告一致初步数据表明，RING1G284A/G284A变体降低了全局H2AUB1级别，导致促凋亡基因的表达增加并减少大脑器官的生长，并破坏双重神经祖细胞（NPC）中的链断裂修复。 AIM 1将探索RING1的遗传机制致病性变体，因为实验会阐明发病机理是否是由于野生型活性丧失引起的 RING1或由于突变等位基因的主要负面影响。 AIM 2将评估分子机制致病环1变体引起小头畸形。实验将评估NPC和类器官与携带RING1致病变体的hESC区分开来，用于DNA损伤修复中的缺陷，增加凋亡的速率以及在DNA损伤部位沉默转录的能力。机械洞察力学从这些实验中，将阐明PRC1依赖性H2AUB1在增殖NPC和IN中的作用。更广泛地维持基因组完整性。此外，这些研究的结果将为未来的努力提供信息制定治疗策略以恢复具有致病性RING1变体的个体的这些功能。