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：
10396109
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/10396109
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prenatal reproductive self-renewal single-cell RNA sequencing sperm cell stem cell differentiation stem cells success unpublished works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.

概括人类生殖细胞对于人类繁殖至关重要，因为只有这些细胞才能够分化进入配子并将DNA从父母传递给孩子。人类生殖系的先锋细胞开始在胎儿期期间形成，当时在胚胎时期留出少量胚胎细胞着床和原肠胚形成的过程称为人类原始生殖细胞（hPGC）规范。这人类生殖细胞发育中的关键事件对个人的未来产生巨大影响生殖健康作为 hPGC 规范的失败会导致某些不孕症。在这场竞争更新中，目标是增加我们对 hPGC 规范的细胞和分子基础的基础知识。基于根据上一资助期的实验结果，我们的目标是使用人类胚胎干细胞（hESC）和人诱导多能干细胞 (hiPSC) 以及 hPGC 样细胞 (hPGCLC) 的分化实现这一目标。总体假设是非啮齿动物和人类特有的分子事件已经进化来调节 hPGC 规范。鉴于这笔拨款的重点主要集中在基因组区域这是人类独有的，该项目非常适合使用基于人类细胞的模型。在目标 1 中，要解决的假设是 TFAP2C 结合的人类特异性逆转录转座子调节 hPGC 规格。在目标 2 中，要解决的假设是 TFAP2C 的表达结合逆转录转座子在 hPGCLC 分化过程中受到表观基因组定向变化的调节。在第三个目标，我们将评估 hPGC 规范中 TFAP2C 和 SOX17 之间的关系，其中假设 TFAP2C 在 hPGC 祖细胞谱系中发挥 SOX17 上游的作用，以调节 hPGC 的规范。总之，这次竞争性续约建立在第一笔融资成功的基础上贡献关于识别人类基因组中新基因座的基本知识的时期进化来调节 hPGC 的规格和身份。