Cellular and Molecular Basis of Human Primordial Germ Cell Specification

人类原始生殖细胞规格的细胞和分子基础

基本信息

批准号：
10226094
负责人：
Amander Clark
金额：
$ 32.87万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-12-20 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10226094
关键词：
Address Animal Model Basic Science Binding CRISPR interference Cell Differentiation process Cell Line Cells ChIP-seq Child Chromatin DNA DNA Methylation Data Development Diagnosis Embryo Endoderm Enhancers Epigenetic Process Event Failure Fertility Fertilization Foundations Funding Future Gametogenesis Gene Expression Genome Germ Cells Goals Gonadal structure Grant Human Human Genome In Vitro Individual Infertility Knowledge Laboratories Life Life Cycle Stages Long Terminal Repeats Modeling Molecular Outcome Parents Process Publishing Reproduction Reproductive Health Research Research Proposals Retrotransposon Role SOX17 gene Scaffolding Protein Site Somatic Cell Specific qualifier value Stem Cell Research Structure of primordial sex cell TFAP2C gene Technology Time To specify Yolk Sac base blastocyst blastomere structure cell type demethylation design egg epigenome gastrulation human embryonic stem cell human pluripotent stem cell implantation induced pluripotent stem cell mutant natural Blastocyst Implantation prenatal reproductive self-renewal single-cell RNA sequencing sperm cell stem cell differentiation stem cells success unpublished works

项目摘要

Summary Human germline cells are essential for human reproduction as only these cells are capable of differentiating into gametes and transmitting DNA from parent to child. The pioneering cells of the human germline begin to form during prenatal life when a small number of embryonic cells are set aside around the time of embryo implantation and gastrulation in a process known as human primordial germ cell (hPGC) specification. This critical event in human germline cell development has a tremendous impact on an individual's future reproductive health as a failure in hPGC specification causes certain infertility. In this competitive renewal, the goal is to increase our fundamental knowledge on the cell and molecular basis of hPGC specification. Based on experimental results in the previous funding period, we aim to use human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) and the differentiation of hPGC-like cells (hPGCLCs) to achieve this goal. The overall hypothesis is that non-rodent and human-specific molecular events have evolved to regulate hPGC specification. Given that the focus of this grant is largely on regions of the genome that are uniquely human, this project is perfectly suited to the use of human cell-based models. In aim 1, the hypothesis to be addressed is that TFAP2C-bound human-specific retrotransposons regulate hPGC specification. In aim 2, the hypothesis to be addressed is that the expression of TFAP2C bound retrotransposons are regulated by targeted changes to the epigenome during hPGCLC differentiation. In the third aim, we will evaluate the relationship between TFAP2C and SOX17 in hPGC specification, with the hypothesis that TFAP2C functions upstream of SOX17 in a lineage primed hPGC progenitor to regulate specification of hPGCs. In summary, this competitive renewal builds upon success from the first funding period to contribute essential knowledge on the identification of new loci in the human genome that have evolved to regulate the specification and identity of hPGCs.

概括人类种系细胞对于人类繁殖至关重要，因为只有这些细胞才能区分进入配子并将DNA从父母传播到孩子。人类种系的开创性细胞开始在产前寿命中形成的形式，当时少量的胚胎细胞在胚胎时搁置一旁在称为人类原始生殖细胞（HPGC）规范的过程中的植入和胃。这人类种系细胞开发中的关键事件对个人的未来产生了巨大影响生殖健康作为HPGC规范的失败会导致某些不育。在这种竞争性更新中目标是增加我们对HPGC规范细胞和分子基础的基本知识。基于在上一个资金期间的实验结果，我们旨在使用人类胚胎干细胞（HESC）和人类诱导的多能干细胞（HIPSC）和HPGC样细胞（HPGCLC）的分化实现这一目标。总体假设是，非变形和人类特异性的分子事件具有演变为调节HPGC规范。鉴于这笔赠款的重点主要是基因组的区域这个项目是独特的人类，非常适合使用基于人类细胞的模型。在AIM 1中要解决的假设是，TFAP2C结合的人类特异性逆转录子调节HPGC 规格。在AIM 2中，要解决的假设是TFAP2C结合的表达在HPGCLC分化过程中，反转座子受到表观基因组的有针对性变化的调节。在第三目的，我们将在HPGC规范中评估TFAP2C和SOX17之间的关系，假设TFAP2C在谱系启动HPGC祖细胞中Sox17上游的功能以调节 HPGC的规范。总而言之，这种竞争性更新以第一笔资金的成功为基础关于在人类基因组中鉴定新基因座的基本知识的时期演变为调节HPGC的规范和身份。