Mechanisms of de novo Germline Histone Mutations Underlying Developmental Disorders

发育障碍背后的从头种系组蛋白突变的机制

基本信息

批准号：
10525879
负责人：
Lijuan Feng
金额：
$ 13.56万
依托单位：
ROCKEFELLER UNIVERSITY
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：
2022-08-25 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10525879
关键词：
ATAC-seq Advisory Committees Affect Affinity Anatomy Animal Model Animals Award Binding Biochemical Biological Assay Brain CRISPR/Cas technology Cell Culture Techniques Cell Nucleus Cell physiology Cells ChIP-seq Child Chromatin Chromatin Structure Clustered Regularly Interspaced Short Palindromic Repeats Cues DNA DNA Repair DNA biosynthesis Data Defect Deposition Development Developmental Biology Embryo Enzymes Epigenetic Process Etiology Exhibits Fibroblasts Fluorescent in Situ Hybridization Future Gene Expression Genes Genomic Imprinting Genomic Segment Genomic approach Genomics Germ-Line Mutation Goals Grant Growth Heterochromatin Higher Order Chromatin Structure Histologic Histology Histone H3 Histone H4 Histones Human Immunohistochemistry In Vitro Intellectual functioning disability Knock-out Knowledge Laboratories Light Lysine Malignant Neoplasms Mass Spectrum Analysis Measures Mentors Mentorship Microcephaly Missense Mutation Mitotic Modeling Modification Molecular Molecular Chaperones Mus Mutant Strains Mice Mutation Neurons Nuclear Nucleosomes Pathologic Pattern Phenotype Physiological Positioning Attribute Process Proteins Protocols documentation Recombinants Research Research Personnel Role Signal Transduction Structural defect Tissues Training Training Support Up-Regulation Variant Whole Organism Work Writing career career development cell fate specification developmental disease embryonic stem cell extracellular imprint in vivo insight migration mouse model mutant nervous system disorder neurogenesis premature programs recruit relating to nervous system single-cell RNA sequencing skills therapeutic candidate transcriptome sequencing zygote

ATAC-seq Advisory Committees Affect Affinity Anatomy Animal Model Animals Award Binding Biochemical Biological Assay Brain CRISPR/Cas technology Cell Culture Techniques Cell Nucleus Cell physiology Cells ChIP-seq Child Chromatin Chromatin Structure Clustered Regularly Interspaced Short Palindromic Repeats Cues DNA DNA Repair DNA biosynthesis Data Defect Deposition Development Developmental Biology Embryo Enzymes Epigenetic Process Etiology Exhibits Fibroblasts Fluorescent in Situ Hybridization Future Gene Expression Genes Genomic Imprinting Genomic Segment Genomic approach Genomics Germ-Line Mutation Goals Grant Growth Heterochromatin Higher Order Chromatin Structure Histologic Histology Histone H3 Histone H4 Histones Human Immunohistochemistry In Vitro Intellectual functioning disability Knock-out Knowledge Laboratories Light Lysine Malignant Neoplasms Mass Spectrum Analysis Measures Mentors Mentorship Microcephaly Missense Mutation Mitotic Modeling Modification Molecular Molecular Chaperones Mus Mutant Strains Mice Mutation Neurons Nuclear Nucleosomes Pathologic Pattern Phenotype Physiological Positioning Attribute Process Proteins Protocols documentation Recombinants Research Research Personnel Role Signal Transduction Structural defect Tissues Training Training Support Up-Regulation Variant Whole Organism Work Writing career career development cell fate specification developmental disease embryonic stem cell extracellular imprint in vivo insight migration mouse model mutant nervous system disorder neurogenesis premature programs recruit relating to nervous system single-cell RNA sequencing skills therapeutic candidate transcriptome sequencing zygote

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项目摘要

Project Summary Chromatin integrates environmental and intrinsic cellular cues to orchestrate nucleosome modifications, therefore regulating basic cellular functions that are essential for cell fate and identity in normal development. Mutations in enzymes that deposit or remove nucleosome modifications often dysregulate chromatin structure and result in pathological gene expression programs in many human developmental disorders. As the basic unit of the nucleosome, alterations in histone genes themselves have only been recently identified in children with developmental disorders and their mechanistic and functional roles remain largely unknown. Given the increasing number of histone germline mutations and lack of understanding of their impact, it is imperative to establish animal models to elucidate their physiological functions and delineate underlying mechanisms. My goal is to uncover the function of histone mutations during development by integrating biochemical and genomic assays, along with employing animal models through the followings aims: (1) Investigate the mechanism of how histone H4 mutants are recruited to heterochromatin, (2) Determine how histone H4 mutations regulate chromatin accessibility and neural differentiation, and (3) Identify the function of histone mutations during development. The central hypothesis guiding this proposal is that histone mutations alter heterochromatin silencing, impact gene expression, and promote neural differentiation, which altogether contribute to brain defects. This research will provide new insights into molecular mechanisms underlying histone germline mutations and the epigenetic causes of developmental disorders. During the mentored period, I will gain training in the following key skillsets: acquiring expertise in mouse models, expanding my knowledge of mouse brain development and in vivo brain models of developmental disorders, deepening training in grant writing and mentoring, as well as scientific career development. With acquisition of these valuable skills, the well-established biochemical and genomics approaches in the Allis laboratory, the great training in neurogenesis and mammalian brain development from my co-mentor Dr. Hatten, and strong support and expertise from my outstanding collaborators, I will be in a unique position to apply diverse approaches to study histone germline mutations in developmental disorders. Importantly, I will receive additional mentoring from my Scientific Advisory Committee, along with fantastic mentorship from Dr. Allis and Dr. Hatten to facilitate my transition to independence. Together, this training and support from the K99/R00 award will fulfill my career goal of becoming an independent investigator in the field of chromatin and developmental biology.

项目摘要染色质整合环境和内在的细胞提示，以编排核小体修饰，因此，调节对正常发育中细胞命运和身份必不可少的基本细胞功能。沉积或去除核小体修饰的酶突变通常会使染色质结构失调并导致许多人类发育障碍中的病理基因表达程序。作为基本单位在核小体中，最近才发现组蛋白基因本身的改变发育障碍及其机械性和功能作用在很大程度上仍然未知。鉴于组蛋白种系突变的数量增加，对其影响不足，必须建立动物模型以阐明其生理功能并描述潜在的机制。我的目标是通过整合生化和基因组来揭示发育过程中组蛋白突变的功能测定以及通过以下内容采用动物模型的目的：（1）研究如何组蛋白H4突变体被募集到异染色质，（2）确定组蛋白H4突变如何调节染色质的可及性和神经分化，（3）确定组蛋白突变的功能发展。指导该建议的中心假设是组蛋白突变改变异染色质沉默，影响基因表达并促进神经分化，这完全有助于大脑缺陷。这项研究将为组蛋白种系的分子机制提供新的见解突变和发育障碍的表观遗传原因。在指导期间，我将获得培训在以下关键技能中：获取鼠标模型中的专业知识，扩大我对鼠标大脑的了解发育和体内大脑的发育障碍模型，加深赠款写作和指导以及科学职业发展。通过获得这些有价值的技能，建立了良好的技能艾里斯实验室的生化和基因组学方法，神经发生和哺乳动物的良好训练我的同事Hatten博士的大脑开发，以及我杰出的强大支持和专业知识合作者，我将处于独特的位置，以应用多种方法来研究组蛋白种系突变发育障碍。重要的是，我将获得科学咨询委员会的其他指导，除了艾利斯博士和哈滕博士的出色指导外，还可以促进我向独立的过渡。一起， K99/R00奖的培训和支持将实现我成为独立的职业目标染色质和发育生物学领域的研究者。