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.

项目摘要摘要部分原因是社会经济原因，在工业化中，父母年龄增加的一般趋势国家。已知高级父母年龄与几个风险因素有关基因组完整性，器官维持受损，突变增加（可能导致癌症），改变了种系中的表观基因组，生育能力降低。重要的是，这些种系改变可以传播到后代，导致多种疾病/综合征的风险更高（例如，肌浮肿，apert，Noonan 和Costello）。但是，人睾丸衰老的详细分子机制是高度理解。关于人类种系和利基市场的先前工作涉及物理方法，但已有未揭示种系发育的详细分子和基因组驱动因素，或者哪些方面详细在衰老期间。强大的单细胞基因组学方法的最近冒险使我们能够检查基因表达，染色质和DNA甲基化（DNAME）全基因组，并为A提供了主要的新机会分子和机械理解衰老对种系和体细胞细胞的影响。该建议是通过结合单细胞基因组，分子和细胞生物学方法的组合，我们的目标为了了解人类种系如何应对衰老的利基市场，以及这是如何反应的与个人的健康和后代有关。我们先前的研究确定了种系的许多原则转录 - 观测关系和继承，提供了青春期和成人的第一个单细胞分析睾丸，并描述了精子dname的第一个年龄依赖性变化。该项目旨在大大扩展我们先前的工作，使用我们的强大睾丸采集管道（保留了66对整个测试获取）检查年轻人通过高龄的年轻人的数千种种系和小众细胞，测试以下假设：人类男性的转录，开放染色质和dname状态衰老期间种系（精子和开发游戏）变化，并对变化做出反应老化的利基。具体而言，我们将利用单细胞基因组学技术检查转录，打开来自年龄分层的男性（20-29、30-39等）的染色质和DNAME谱。我们的目标了解衰老对种系的影响（AIM 1）和体细胞细胞（AIM 2）。重要的是，我们会的还采用功能实验，包括精子症异种移植和体外器官培养物，旨在测试种系或验证细胞的发育潜力。此外，作为BMI（体重）索引）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，因为我们已经有足够的样品将每个年龄范围分类为高和低BMI。通过确定潜在的驱动因素来在一起对于人类睾丸衰老，我们旨在改变与年龄相关病理的管理（例如不孕症和癌症）并获得了对由晚期父亲年龄引起的疾病/疾病的深刻分子理解。