Measurement of cell-type specific somatic mutation rates in the aging human brain - Resubmission – 1

衰老人脑中细胞类型特异性体细胞突变率的测量 - 重新提交 ‐ 1

• 批准号: 10731067
• 负责人: Marta Gronska-Peski
• 金额: $ 7.18万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10731067
• 关键词: Address; Age; Aging; Alzheimer' s Disease; Antibodies; Astrocytes; Autopsy; Award; Bioinformatics; Brain; Brain Diseases; Brain Neoplasms; Brain region; Career Mobility; Cell Nucleus; Cell Separation; Cells; Collaborations; Communities; DNA; DNA Repair; DNA biosynthesis; DNA sequencing; Dedications; Disease; Dissociation; Doctor of Philosophy; Elderly; Environment; Flow Cytometry; Fluorescent in Situ Hybridization; Formalin; Foundations; Freezing; Future; Genetic; Genetic Predisposition to Disease; Genome; Genomics; Goals; Human; Human Genetics; Human body; Individual; Interneurons; Knowledge; Label; Laboratories; Libraries; Longevity; Measurement; Measures; Mentorship; Metabolic; Methods; Molecular; Molecular Profiling; Morbidity - disease rate; Mutagenesis; Mutation; Neurodegenerative Disorders; Neuroglia; Neurons; Neurosciences; Nuclear Antigens; Organ; Pathogenesis; Pathologic Mutagenesis; Pathology; Pattern; Preparation; Process; Protocols documentation; Reaction; Reactive Oxygen Species; Research; Role; Scientific Advances and Accomplishments; Somatic Mutation; Surface Antigens; System; Techniques; Tissues; Toxin; Training; Work; age group; age related; aging brain; brain cell; brain tissue; career; career development; cell type; crosslink; genome integrity; human genomics; human tissue; medical schools; mortality; new technology; novel; phosphoproteomics; preservation; sample fixation; single nucleus RNA-sequencing; statistics; stem; success; technology platform; transcriptome sequencing

Project summary/Abstract Research: The human brain contains over 100 diverse classes and subclasses of neurons and glia, making it the most heterogenous organ in the body. Recent studies have revealed that during aging, brain cells accumulate somatic mutations with patterns that reflect diverse mutagenic factors. However, to date, these studies have been unable to systematically profile aging-related somatic mutation rates (SMRs) of specific brain cell types due to the lack of methods to isolate specific cell types from human brain tissue without the process of fixation, which precludes high-fidelity sequencing necessary for measuring SMRs. This proposal aims to fill this knowledge gap by developing a method to isolate potentially any cell type from post-mortem human brain, while avoiding fixation in order to preserve the integrity of the genome for high-fidelity DNA- sequencing (DNA-seq) and analysis of SMRs. Understanding how SMRs and mutational signatures differ among cell types in the brain is important, because it is a prerequisite for assessing the possible role of somatic mutations and genome integrity in age-related brain diseases that often involve specific cell types. The proposal’s aims are: 1) To develop a novel technology to isolate unfixed nuclei of any cell type from frozen post-mortem human brains, a method which will also potentially be applicable to any tissue; 2) To determine SMRs across age spans for four specific human brain cell types relevant to age-related brain diseases. Successful completion of this proposal will enable future studies of somatic mutations and other molecular profiling that is precluded by fixation in potentially any cell type and brain region in the context of diverse age- related brain disorders. Training Plan: Under this award, I will 1) Expand my scientific knowledge of the mechanisms of mutagenesis and genome integrity generally, and in the context of the aging human brain; 2) Gain computational and bioinformatics expertise in genomics and DNA-seq analysis; 3) Conduct career development activities for successful transition towards a future independent academic research career. Environment: The proposed research and training plans will be conducted in the laboratory of Dr. Gilad Evrony, MD, PhD, at the Center for Human Genetics and Genomics (CHGG) at NYU Grossman School of Medicine (NYUSOM), with co-mentorship from the Center’s director, Dr. Aravinda Chakravarti, PhD. NYUSOM is an environment with well-respected and established research groups dedicated to trainees’ success. Dr. Evrony is a pioneer and expert in single-cell genomics and computational genomics, providing a rigorous and supportive research environment along with career mentorship. Dr. Chakravarti is a renowned expert in human genetics and genomics, as well as statistics, who will provide further guidance on scientific, computational, and career goals. Both laboratories, the collaborating research community, and our affiliations with the Neuroscience and Systems Genetics Institutes, provide a unique environment to integrate neuroscience and genomics training, and to expand scientific and career-advancement activities to aid my transition towards independence.

项目摘要/摘要 研究：人脑包含100多种潜水员类和神经元和神经胶质的子类 体内最异性的器官。最近的研究表明，在衰老期间，脑细胞 用反映潜水诱变因子的模式积累体细胞突变。但是，到目前为止，这些 研究无法系统地介绍特定特定的与衰老相关的体细胞突变率（SMR） 由于缺乏将特定细胞类型与人类组织分离的方法，脑细胞类型没有 固定过程，这排除了测量SMR所需的高保真测序。这个建议 旨在通过开发一种将潜在的任何细胞类型隔离后的方法来填补这一知识空白 人脑，同时避免固定以保持基因组对高保真DNA-的完整性 测序（DNA-SEQ）和SMR分析。了解SMR和突变签名如何不同 在大脑中的细胞类型中很重要，因为它是评估可能作用的先决条件 与年龄相关的脑疾病中的体细胞突变和基因组完整性通常涉及特定的细胞类型。这 提案的目的是：1）开发一种新型技术，以将任何细胞类型的未固定核电从冷冻中分离出来 验尸后人的大脑，这种方法也可能适用于任何组织； 2）确定 跨年龄的SMR跨越了与年龄相关的脑疾病有关的四种特定人类脑细胞类型。 该提案的成功完成将使未来的体细胞突变和其他分子研究 在不同年龄的背景下，在任何细胞类型和大脑区域的固定中所无法进行的分析 - 相关的脑部疾病。培训计划：根据该奖项，我将1）扩大我对 通常，在衰老的人脑的背景下，诱变和基因组完整性的机制； 2） 获得基因组学和DNA-seq分析方面的计算和生物信息学专业知识； 3）从事职业 成功过渡到未来独立学术研究职业的发展活动。 环境：拟议的研究和培训计划将在吉拉德·伊夫里（Gilad Evrony）博士的实验室中进行， 医学博士，博士，在人类遗传与基因组中心（CHGG）的纽约大学格罗斯曼医学院 （Nyusom），该中心主任Aravinda Chakravarti博士的冠军。 Nyusom是一个 具有尊敬和成熟的研究小组的环境致力于学员的成功。 Evrony博士是 单细胞基因组学和计算基因组学领域的先驱和专家，提供了严格而支持的 研究环境以及职业心态。 Chakravarti博士是人类遗传学的著名专家 以及基因组学以及统计数据，他们将提供有关科学，计算和职业的进一步指导 目标。两个实验室，合作研究社区以及我们与神经科学和神经科学的关联 系统遗传学机构，为整合神经科学和基因组学培训提供了独特的环境， 并扩大科学和职业投资活动，以帮助我向独立过渡。