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Protecting and Sustaining Germ Cell Identity

保护和维持生殖细胞身份

基本信息

批准号：
10736134
负责人：
RUTH LEHMANN
金额：
$ 41.93万
依托单位：
WHITEHEAD INSTITUTE FOR BIOMEDICAL RES
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-11 至 2028-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10736134
关键词：
ATAC-seq Address Binding Binding Sites Biological Assay Cell Reprogramming Cells Chromatin Chromatin Remodeling Factor DNA Transposable Elements DNA damage checkpoint Deposition Development Elements Embryo Embryonic Development Enzymes Face Gene Activation Gene Expression Gene Expression Regulation Genes Genetic Genetic Transcription Genome Germ Germ Cells Goals Human Immunity Inherited Longevity Mediating Methods Modeling Molecular Mus Mutation Oogenesis Pathway interactions Phenotype Process Proteins RNA RNA Editing RNA-Binding Proteins Regulation Regulatory Pathway Reporter Repression Role Site Small RNA Somatic Cell Specific qualifier value Structure of primordial sex cell System Testing Totipotency Trans-Activators Transact Transcriptional Activation Transcriptional Regulation Translational Repression Translations Validation candidate identification cell type egg expectation experimental study fly member mutant nano nanobodies neuronal cell body next generation novel permissiveness piRNA preempt premature prevent programs reproductive ribosome profiling single-cell RNA sequencing sperm cell transcription factor transmission process tumor

项目摘要

Project Summary/Abstract Germ cells have the extraordinary potential to generate every cell type in the body. Yet, the molecular program that protects and sustains this promise for totipotency in germ cells, which can last more than forty years in humans, is poorly understood. To face this challenge, germline regulators assume dual roles in maintaining germ cell identity: activating a germline transcriptional program while simultaneously protecting germ cells from reprogramming to a somatic cell fate. Indeed, genes involved in germline specification, such as the conserved RNA-binding protein Nanos and the transposable element (TE) regulator Piwi, are erroneously upregulated in certain human tumors. These tumors are thought to undergo soma-to-germline transformations to acquire a more immortal, germline-like state. However, a specific program for germ cell transcriptional activation has yet to be described, mainly because a ‘master-regulator transcription factor’ for germ cell fate is missing. This proposal uses a multipronged approach to systematically characterize the gene expression program in primordial germ cells (PGCs) and identify prime regulators of germ cell fate. The goal is to distinguish between a ‘default’ model, by which repression of the somatic program ‘allows’ the PGC program, and an ‘instructive’ model, by which PGC specification is actively controlled. In Aim1, we probe the instructive model to uncover the germline transcriptional program. By single-cell RNA sequencing, we have detected specific, temporally-regulated germline genes. To identify the transacting regulatory factors, we will use ATAC-sequencing, characterize known factors with matching binding activities, and identify potential new regulatory candidates for germline genes. In Aim2, we address the function of a critical regulator of germ cell fate, the conserved RNA regulator Nanos. Using the RNA target identification method hyper-TRIBE and ribosome profiling, we will identify Nanos targets and functionally distinguish between those targets that promote the germline program and/or repress the somatic program. Aim3 focuses on the piRNA pathway, which provides maternally inherited immunity against TE activity by controlling the transcription of TE elements, specifically in germ cells. Using a novel degradation strategy to specifically degrade components of the piRNA pathway in PGCs without impacting oogenesis, we will determine the role of this pathway for embryonic PGC gene expression beyond TE control. By integrating multiplex findings, this proposal will uncover the regulatory landscape of early germ cells, thereby providing a necessary understanding of the process of gonadogenesis and, ultimately, the survival of the species.

项目总结/文摘