Admin Supplement: Epigenetic Determinants of Competence for Germ Line Specification during Cynomolgus Embryonic Development

管理补充：食蟹猴胚胎发育过程中种系规格能力的表观遗传决定因素

• 批准号: 10704459
• 负责人: Werner Neuhausser
• 金额: $ 5.4万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10704459
• 关键词: Affect; Animal Model; Area; Award; Bioinformatics; Biological Assay; Biology; Cells; Cellular biology; Clustered Regularly Interspaced Short Palindromic Repeats; Competence; Congenital Abnormality; Defect; Development; Developmental Biology; Diabetes Mellitus; Disease; Drug Addiction; Embryo; Embryonic Development; Epigenetic Process; Future; Gametogenesis; Genetic; Genetic study; Genomics; Germ; Germ Cells; Germ Lines; Goals; Grant Review; Hospitals; Human; In Vitro; Infertility; Institution; Investigation; Israel; Knowledge; Late-Onset Disorder; Link; Malignant Neoplasms; Mediating; Medical center; Mentors; Mesoderm; Mission; National Institute of Child Health and Human Development; Outcome; Physicians; Pluripotent Stem Cells; Pregnancy loss; Regulator Genes; Regulatory Element; Reporter; Reproductive Endocrinology; Research; Research Personnel; Research Priority; Somatic Cell; Specific qualifier value; Structure of primordial sex cell; Training; United States National Institutes of Health; Woman; catalyst; differentiation protocol; epigenome editing; epigenomics; experience; experimental study; gastrulation; gene regulatory network; human disease; improved; instructor; laboratory experience; loss of function; multidisciplinary; pluripotency; programs; stem cell biology; stem cells; tool; transdifferentiation

Project Summary CANDIDATE. I am a physician and Instructor in the Division of Reproductive Endocrinology and Infertility at Beth Israel Deaconess Medical Center (BIDMC). A K08 Award will provide me with the necessary training and experience to become an independent investigator in germ cell (GC) biology, with a focus on the gene regulatory networks underlying human gametogenesis. BACKGROUND. How a small subset of cells in the developing human embryo acquires competency to form the germ line and ultimately mature into functional gametes while maintaining its potential for pluripotency is among the least understood questions in biology. In particular, the gene regulatory networks and epigenetic remodeling, which govern competence for the GC fate during specification of early (primordial) germ cells (PGCs) in the peri-gastrulating period, remain unknown. Knowledge of the underlying mechanisms will allow the development of more efficient differentiation protocols for GCs from pluripotent stem cells (PSCs) in vitro. In addition, it will allow the investigation of the link between epigenetic dysregulation during GC specification and critical human disorders such as pregnancy loss, congenital defects and common late-onset diseases such as infertility, diabetes, drug addiction and cancer. RESEARCH. The objective for this K08 is to identify and validate the key gene regulatory elements (GREs) governing induction of competence for the GC fate and the differentiation of PGCs during human gastrulation. Our central hypothesis is that epigenetic priming of key GREs in incipient mesoderm mediates developmental competence during germ lineage specification from the pluripotent state. This research will pursue two specific aims: (1) to identify putative GREs underlying developmental competence for the GC fate through epigenetic footprinting (2) to identify the subset of these GREs functioning as key determinants of GC competence and differentiation using massively parallel reporter assays (MPRA) and CRISPR epigenome editing mediated loss of function experiments. In the future, these results will provide the platform to use epigenome editing as a tool to improve the efficiency of GC specification, for trans-differentiation of GCs from somatic cells and to study the effect of epimutations on developmental outcomes in GCs and embryos using animal models. MENTORING. My primary mentor, Dr. Raymond Anchan, is an expert in stem cell biology at Brigham and Women’s Hospital. I have assembled a multi-disciplinary mentoring team across the Harvard network: Dr. Alex Meissner (an expert in epigenetics and epigenomics), Dr. Steve McCarroll (a leader in genomics and bioinformatics). TRAINING. The research objectives are supported by a training plan that includes laboratory training as well as rigorous formal didactics in genomics, bioinformatics, epigenetics and developmental biology alongside a strong institutional commitment at BIDMC and the grant review and support program through Harvard Catalyst.

项目摘要 候选人。我是生殖内分泌学和不孕不育科的内科医生和讲师 贝丝以色列女执事医疗中心(BIDMC)。K08奖将为我提供必要的培训和 有成为生殖细胞(GC)生物学独立研究员的经验，专注于基因研究 人类配子发生的基础调控网络。 背景资料。发育中的人类胚胎中的一小部分细胞如何获得形成能力 胚系并最终成熟为功能配子，同时保持其多能性的潜力是 这是生物学中最不被理解的问题之一。特别是，基因调控网络和表观遗传学 重构，在早期(原始)生殖细胞的指定过程中控制GC命运的能力 (Pgcs)在周原肠形成期，仍然未知。对潜在机制的了解将使 从多能干细胞(PSCs)体外分化为GCs的更有效方案的开发。 此外，它还将允许研究GC规范过程中表观遗传失调之间的联系 以及严重的人类疾病，如流产、先天缺陷和常见的晚发性疾病 如不孕不育、糖尿病、毒瘾和癌症。 研究。本K08的目标是识别和验证关键基因调控元件(Gres)。 在人类原肠发育过程中，对GC命运和PGCs分化的能力的诱导。 我们的中心假设是，早期中胚层关键GRE的表观遗传启动介导了发育 多能状态下的生殖细胞谱系确定能力。这项研究将追求两个具体的 目的：(1)通过表观遗传学确定GC命运潜在的GRE发育能力 足迹(2)以确定这些GRE的子集作为GC能力的关键决定因素以及 大规模平行报告分析(MPRA)和CRISPR表观基因组编辑介导的丢失 机能实验。在未来，这些结果将为使用表观基因组编辑作为工具提供平台 为了提高GC分型的效率，提高体细胞转分化的效率，并研究 用动物模型研究突变对GC和胚胎发育结果的影响。 辅导。我的主要导师雷蒙德·安昌博士是布里格姆大学的干细胞生物学专家 女子医院。我已经组建了一个跨哈佛网络的多学科指导团队：Alex博士 迈斯纳(表观遗传学和表观基因组学专家)，史蒂夫·麦卡罗尔博士(基因组学和 生物信息学)。 训练。研究目标得到了包括实验室培训在内的培训计划的支持 作为基因组学、生物信息学、表观遗传学和发育生物学的严格正式教学学，以及 BIDMC坚定的机构承诺，以及通过哈佛催化剂的赠款审查和支持计划。