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.

项目概要 我是生殖内分泌和不孕不育科的医生和讲师。 贝斯以色列女执事医疗中心 (BIDMC) 将为我提供必要的培训和帮助。 拥有成为生殖细胞 (GC) 生物学独立研究者的经验，重点研究基因 人类配子发生的调控网络。 背景：发育中的人类胚胎中的一小部分细胞如何获得形成的能力。 种系并最终成熟为功能性配子，同时保持其多能性潜力 特别是基因调控网络和表观遗传学是生物学中最不为人所知的问题之一。 重塑，在早期（原始）生殖细胞规范过程中控制 GC 命运的能力 （PGC）在围原肠胚形成期间，仍然未知的潜在机制将允许。 开发更有效的多能干细胞 (PSC) 体外分化 GC 方案。 此外，它还可以研究 GC 规范期间表观遗传失调之间的联系 以及严重的人类疾病，例如流产、先天性缺陷和常见的迟发性疾病 例如不孕症、糖尿病、毒瘾和癌症。 研究。K08 的目标是识别和验证关键基因调控元件 (GRE)。 控制人类原肠胚形成过程中 GC 命运和 PGC 分化能力的诱导。 我们的中心假设是，早期中胚层中关键 GRE 的表观遗传启动介导发育 这项研究将追求两个特定的能力。 目标：(1) 通过表观遗传学确定 GC 命运的潜在发育能力的推定 GRE 足迹 (2) 来识别这些 GRE 的子集，作为 GC 能力的关键决定因素， 使用大规模并行分析报告编辑 (MPRA) 和 CRISPR 表观基因组介导的丢失进行分化 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。 提高 GC 规范的效率，从体细胞转分化 GC，并研究 使用动物模型研究表观突变对 GC 和胚胎发育结果的影响。 我的主要导师 Raymond Anchan 博士是布莱根医学院的干细胞生物学专家。 我在哈佛网络组建了一支多学科指导团队：亚历克斯博士。 Meissner（表观遗传学和表观基因组学专家）、Steve McCarroll 博士（基因组学和表观基因组学领域的领导者） 生物信息学）。 培训 研究目标得到培训计划的支持，其中还包括实验室培训。 作为基因组学、生物信息学、表观遗传学和发育生物学方面严格的正式教学法，以及 BIDMC 的强有力的机构承诺以及通过哈佛催化剂的资助审查和支持计划。