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

• 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
• 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

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.

项目概要 染色质整合环境和内在细胞线索来协调核小体修饰， 因此调节对正常发育中的细胞命运和身份至关重要的基本细胞功能。 沉积或去除核小体修饰的酶突变通常会导致染色质结构失调 并导致许多人类发育障碍的病理性基因表达程序。作为基本单位 在核小体中，组蛋白基因本身的改变直到最近才在患有 发育障碍及其机制和功能作用仍然很大程度上未知。鉴于 组蛋白种系突变数量不断增加，但对其影响缺乏了解，因此必须 建立动物模型以阐明其生理功能并描绘潜在机制。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 分析，并通过以下目标采用动物模型：（1）研究如何作用的机制 组蛋白 H4 突变体被招募到异染色质，（2）确定组蛋白 H4 突变如何调节 染色质可及性和神经分化，以及（3）鉴定组蛋白突变的功能 发展。指导该提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，促进神经分化，这些都有助于大脑 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。在辅导期间，我将获得培训 在以下关键技能方面：获得小鼠模型方面的专业知识，扩展我对小鼠大脑的了解 发育和发育障碍的体内脑模型，深化资助写作和 指导以及科学的职业发展。通过获得这些宝贵的技能，成熟的 艾利斯实验室的生物化学和基因组学方法，神经发生和哺乳动物方面的良好培训 我的共同导师 Hatten 博士的大脑开发，以及我杰出的人的大力支持和专业知识 作为合作者，我将处于一个独特的位置，可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，我将从我的科学咨询委员会获得额外的指导， 以及艾利斯博士和哈滕博士的出色指导，帮助我过渡到独立。一起， K99/R00 奖项的培训和支持将实现我成为独立人士的职业目标 染色质和发育生物学领域的研究员。