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

基本信息

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
项目类别：
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&apos 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

项目摘要

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.

项目概要/摘要研究：人类大脑包含 100 多个不同类别和亚类的神经元和神经胶质细胞，使其最近的研究表明，脑细胞是体内最异质的器官。然而，迄今为止，这些体细胞突变的模式反映了不同的诱变因素。研究无法系统地分析与衰老相关的特定体细胞突变率（SMR）由于缺乏从人脑组织中分离特定细胞类型的方法而无法识别脑细胞类型固定过程，这妨碍了测量 SMR 所需的高保真测序。旨在通过开发一种从死后分离潜在任何细胞类型的方法来填补这一知识空白人脑，同时避免固定以保持高保真 DNA 基因组的完整性了解 SMR 和突变特征的不同之处。大脑细胞类型之间的差异很重要，因为它是评估大脑细胞类型可能作用的先决条件与年龄相关的脑部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。该提案的目标是：1）开发一种新技术，从冷冻细胞中分离任何细胞类型的未固定细胞核死后人类大脑，这种方法也可能适用于任何组织；2) 确定与年龄相关的脑部疾病相关的四种特定人类脑细胞类型的跨年龄跨度 SMR。该提案的成功完成将使未来对体细胞突变和其他分子的研究成为可能。在不同年龄的背景下，通过固定可能的任何细胞类型和大脑区域来排除分析相关的脑部疾病的培训计划：根据这个奖项，我将 1）扩展我的科学知识。诱变和基因组完整性的一般机制，以及在人类大脑衰老的背景下；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) 开展职业生涯；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 Gilad Evrony 博士的实验室进行，纽约大学格罗斯曼医学院人类遗传学和基因组学中心 (CHGG) 医学博士、博士 (NYUSOM)，由该中心主任 Aravinda Chakravarti 博士共同指导。 Evrony 博士拥有受人尊敬且知名的研究小组，致力于帮助学员取得成功。单细胞基因组学和计算基因组学的先驱和专家，提供了严格和支持性的 Chakravarti 博士是人类遗传学领域的著名专家。和基因组学以及统计学，他们将为科学、计算和职业提供进一步的指导两个实验室、合作研究团体以及我们与神经科学和神经科学的关系。系统遗传学研究所提供了整合神经科学和基因组学培训的独特环境，并扩大科学和职业发展活动，以帮助我向独立过渡。