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

• 批准号: 10613472
• 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
• 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/10613472
• 关键词: Address; 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; 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; Mus; 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; Yolk Sac; blastocyst; blastomere structure; cell type; demethylation; design; egg; embryo cell; epigenome; gastrulation; human embryonic stem cell; human pluripotent stem cell; implantation; induced pluripotent stem cell; model organism; mutant; natural Blastocyst Implantation; prenatal; reproductive; self-renewal; single-cell RNA sequencing; sperm cell; stem cells; success; transmission process; unpublished works

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。

项目成果

Standing on the shoulders of giants: The changing landscape of pluripotent stem cells in research.

站在巨人的肩膀上：多能干细胞研究中不断变化的景观。

• 发表时间: 2020
• 作者: Clark; Amander T
• 通讯作者: Amander T

Human Primordial Germ Cells Are Specified from Lineage-Primed Progenitors

人类原始生殖细胞是由谱系引发的祖细胞特化而来

• DOI: 10.1109/jqe.2008.2002097
• 发表时间: 2009-02-01
• 期刊: IEEE Journal of Quantum Electronics
• 影响因子: 2.5
• 作者: Di Chen;Lei Hou .. Na Sun;Wanlu Liu;M. Kellis;Am;er T Clark;er;Lei Hou;Rachel Kim;Jared Faith;M. Aslanyan;Yu Tao;Yi Zheng;Jianping Fu
• 通讯作者: Jianping Fu

Divergent roles for KLF4 and TFCP2L1 in naive ground state pluripotency and human primordial germ cell development.

KLF4 和 TFCP2L1 在幼稚基态多能性和人类原始生殖细胞发育中的不同作用。

• DOI: 10.21203/rs.3.rs-24873/v1
• 发表时间: 2020-04-24
• 期刊: Stem cell research
• 影响因子: 1.2
• 作者: Grace V. Hancock;Wanlu Liu;Lior Peretz;Di Chen;Joanna J. Gell;A. Collier;Jesse R Zamudio;K. Plath;A. Clark
• 通讯作者: A. Clark

Modeling human infertility with pluripotent stem cells.

用多能干细胞模拟人类不育症。

• DOI: 10.1016/j.scr.2017.04.005
• 发表时间: 2017-05
• 期刊: Stem cell research
• 影响因子: 1.2
• 作者: Chen D;Gell JJ;Tao Y;Sosa E;Clark AT
• 通讯作者: Clark AT

The TFAP2C-Regulated OCT4 Naive Enhancer Is Involved in Human Germline Formation.

TFAP2C 调节的 OCT4 初始增强子参与人类种系形成。

• 发表时间: 2018
• 影响因子: 8.8
• 作者: Chen, Di;Liu, Wanlu;Zimmerman, Jill;Pastor, William A;Kim, Rachel;Hosohama, Linzi;Ho, Jamie;Aslanyan, Marianna;Gell, Joanna J;Jacobsen, Steven E;Clark, Amander T
• 通讯作者: Clark, Amander T