Molecular, Epigenetic and Genomic Approaches to Understand Mechanisms of Aging in the Human Testis

通过分子、表观遗传学和基因组方法了解人类睾丸衰老机制

基本信息

批准号：
10646506
负责人：
BRADLEY R. CAIRNS
金额：
$ 38.13万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-30 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10646506
关键词：
ATAC-seq Achondroplasia Adult Age Aging Autopsy Biological Assay Body mass index Cells Cellular biology Child Chromatin DNA Methylation DNA methylation profiling Data Data Set Deterioration Developed Countries Development Disease Elderly Endothelium Epigenetic Process Fertility Gene Expression Genetic Genetic Transcription Genomic approach Genomics Germ Cells Health Hereditary Disease Human Impairment In Vitro Individual Infertility Link Macrophage Maintenance Malignant Neoplasms Malignant neoplasm of testis Molecular Molecular Biology Morphology Mutation Nucleic Acid Regulatory Sequences Organ Organoids Parental Ages Paternal Age Pathology Pattern Physiological Physiology Public Health Reproductive Health Risk Risk Factors Sampling Series Spermatids Spermatocytes Spermatogonia Structure Syndrome System Techniques Testing Testis Tissues Validation Work Xenograft procedure age related age stratification aged bioinformatics pipeline cell age epigenome experimental study genome integrity genome wide methylation high body mass index high risk human male male men offspring preservation reproductive single cell analysis single-cell RNA sequencing socioeconomics sperm cell stem cells transcriptome sequencing transmission process trend young adult

项目摘要

PROJECT SUMMARY ABSTRACT Due partly to socio-economic reasons, there is a general trend of increased parental age in the industrialized countries. Advanced parental age is known to be associated with several risk factors including decreased genome integrity, impaired organ maintenance, increased mutations (which may cause testicular cancer), altered epigenome in the germline, and reduction in fertility. Importantly, these germline alterations can be transmitted to the offspring, resulting in higher risks for several diseases/syndromes (e.g. Achondroplasia, Apert, Noonan and Costello) in their children. However, the detailed molecular mechanism underlying human testis aging is highly understudied. Prior work on human germline and niche has involved physiological approaches, but has not revealed the detailed molecular and genomic drivers of germline development, or which aspects deteriorate during aging. The recent advent of powerful single-cell genomics approaches allows us to examine gene expression, chromatin and DNA methylation (DNAme) genome-wide, and offer major new opportunities for a molecular and mechanistic understanding of the impact of aging on both germline and somatic niche cells. In this proposal, by utilizing a combination of single cell genomics, molecular and cell biology approaches, we aim to gain a mechanistic understanding of how human germline ages in respond to the aging niche, and how this is related to individuals’ health and that of their offspring. Our prior studies established many principles of germline transcription-epigenetic relationships and inheritance, provided the first single-cell analyses of pubertal and adult testes, and described the first age-dependent changes in sperm DNAme. This project aims to greatly extend our prior work, by utilizing our robust testis acquisition pipeline (66 pairs of whole testes preserved with ongoing acquisition) to examine thousands of germline and niche cells from young adults through men of advanced age, to test the following Hypothesis: The transcription, open chromatin and DNAme status of human male germline (spermatogonia and developing gametes) changes during aging, and responds to changes in the aging niche. Specifically, we will utilize single cell genomics techniques to examine the transcription, open chromatin and DNAme profiles of testicular cells from age stratified males (20-29, 30-39 etc). We aim to understand the impact of aging on the germline (AIM 1) and somatic niche cells (AIM 2). Importantly, we will also apply functional experiments including spermatogonia xenotransplantation and in vitro organoid culturing, aiming to test the developmental potential of germline or testicular niche cells. Furthermore, as BMI (body mass index) often increases with age, we will also assess the correlations of observed changes with BMI, as we have sufficient samples to stratify each age range by high and low BMI. Taken together, by identifying potential drivers for human testis aging, we aim to transform the management of age-associated pathology (e.g. infertility and cancer) and gain a deep molecular understanding of the diseases/disorders caused by advanced paternal age.

项目概要摘要部分由于社会经济原因，工业化国家普遍存在父母年龄增长的趋势。众所周知，父母高龄与多种风险因素有关，包括年龄下降。基因组完整性、器官维护受损、突变增加（可能导致睾丸癌）生殖系的表观基因组和生育能力的降低重要的是，这些生殖系的改变可以传播。给后代带来更高的风险，导致多种疾病/综合征（例如软骨发育不全、Apert、Noonan 和科斯特洛）在他们的孩子中然而，人类睾丸衰老的详细分子机制是先前对人类种系和生态位的研究涉及生理学方法，但尚未得到充分研究。没有揭示种系发育的详细分子和基因组驱动因素，或者哪些方面恶化最近出现的强大的单细胞基因组学方法使我们能够检查衰老过程中的基因。基因组范围内的表达、染色质和 DNA 甲基化 (DNAme)，并为从分子和机制上理解衰老对生殖系和体细胞生态位细胞的影响。该提案通过结合单细胞基因组学、分子和细胞生物学方法，我们的目标是获得对人类种系衰老如何响应衰老生态位的机制性理解，以及这是如何发生的我们之前的研究建立了许多种系原则。转录-表观遗传关系和遗传，首次提供了青春期和成人的单细胞分析测试，并描述了精子 DNAme 的第一个年龄依赖性变化。该项目旨在极大地扩展我们的研究。之前的工作，通过利用我们强大的睾丸采集管道（66 对完整测试，持续进行保存）采集）来检查从年轻人到老年男性的数千种生殖细胞和生态位细胞，测试以下假设：人类男性的转录、开放染色质和 DNAme 状态种系（精原细胞和发育中的配子）在衰老过程中发生变化，并对生殖细胞的变化做出反应具体来说，我们将利用单细胞基因组学技术来检查转录，打开。我们的目标是对年龄分层男性（20-29、30-39 等）的睾丸细胞进行染色质和 DNAme 分析。了解衰老对生殖细胞 (AIM 1) 和体细胞生态位细胞 (AIM 2) 的影响。还应用精原细胞异种移植和体外类器官培养等功能实验，旨在测试生殖细胞或睾丸微环境细胞的发育潜力此外，BMI（体重）。指数）通常随着年龄的增长而增加，我们还将评估观察到的变化与 BMI 的相关性，因为我们有通过确定潜在的驱动因素，将足够的样本按高体重指数和低体重指数对每个年龄范围进行分层。对于人类睾丸衰老，我们的目标是改变与年龄相关的病理学的管理（例如不孕症和癌症），并深入了解高龄父亲造成的疾病/紊乱。