FA DDR Pathway in Germline Integrity

种系完整性中的 FA DDR 途径

• 批准号: 9918948
• 负责人: QISHEN PANG
• 金额: $ 32.99万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-11 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9918948
• 关键词: Apoptosis; Area; Binding Proteins; Biochemical; Biochemical Genetics; Clinical; Collaborations; Congenital Abnormality; DNA Damage; DNA Double Strand Break; DNA Methylation; DNA Repair; DNA Transposable Elements; Defect; Development; Disease; Embryonic Development; Epigenetic Process; Fanconi Anemia Complementation Group A Protein; Fanconi Anemia pathway; Fanconi' s Anemia; Fertility; Genetic; Genetic Materials; Genome; Genomic Instability; Germ Cells; Goals; Health; Health Technology; Hemagglutinin; Infertility; Investigation; Knock-in Mouse; Knockout Mice; Knowledge; Lead; Link; Maintenance; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Modeling; Molecular; Mus; National Institute of Child Health and Human Development; Nuclear; Oogenesis; Organ; Pancytopenia; Pathway interactions; Patients; Predisposition; Proteins; Public Health; Repression; Reproduction; Research Design; Role; Signal Transduction; Spermatogenesis; Structure of primordial sex cell; Syndrome; Testing; Therapeutic; Transcriptional Silencer Elements; Up-Regulation; chromatin modification; early embryonic stage; egg; fertility preservation; genetic approach; improved; in vivo; innovation; mouse model; next generation; offspring; piRNA; precursor cell; prevent; programs; repaired; reproductive; response; spatiotemporal; sperm cell

Abstract Genome maintenance in the germline is vital for fertility and the health of offspring. Although DNA damage response (DDR) and repair has been well characterized in meiocytes, the DDR pathway(s) that function in primordial germ cells (PGCs), which are the precursors to sperm and eggs, have not been identified. Fanconi anemia (FA) is a genomic instability syndrome associated with PGC depletion and infertility in addition to some other devastating clinical manifestations such as bone marrow failure and cancer predisposition. The FA pathway is a major DDR pathway known to function in double-strand DNA break repair. The signaling mechanisms underlying PGC depletion and infertility in FA patients or mouse models are not known. We recently developed Flag- and hemagglutinin-tagged Fancd2 knock-in mice that allowed a high throughput mass spectrometry approach to search for Fancd2-binding proteins in different mouse organs. In addition to DDR partners, we observed several proteins of the germ-cell-specific Prmt5/piRNA pathways orchestrating the repression of transposable elements (TEs). Deletion of Fancd2 resulted in decreased protein levels of the Prmt5/piRNA factors, massive upregulation of TEs, PGC depletion, and led to defective spermatogenesis and oogenesis in Fancd2-null (Fancd2-KO) mice. These preliminary studies suggest that in addition to its well-established DNA repair roles, Fancd2 and the FA pathway has an in vivo TE silencing function in early-stage germ cells. We hypothesize that the FA pathway is essential for germline integrity involving a pathway hierarchy in which the FA core signals to Fancd2, which then guides Prmt5 and piRNA in TE silencing. The goals of the project are to establish functional interaction between the FA pathway and germ cell- specific TE silencing machinery in safeguarding the germline genome, and to define Fancd2 as a crucial regulator of this vital epigenetic programming during germ cell development. The project presents an innovative study aimed at linking a major DDR pathway to germline genome maintenance in early-stage germ cells. The knowledge gained from the proposed study will not only improve mechanistic understanding of the molecular collaboration between the FA DDR pathway and the Prmt5/piRNA pathway in the context of TE repression, but also lead to a new avenue of research designed to target these interacting pathways for developing innovative therapeutic strategies for reproductive diseases such as infertility and birth defects.

抽象的 种系中的基因组维持对于生育能力和后代的健康至关重要。虽然DNA 损伤响应（DDR）和维修在Meiocytes中的表征很好，DDR 在原始生殖细胞（PGC）中起作用的途径，这是精子和精子的前体 鸡蛋，尚未确定。 Fanconi贫血（FA）是一种基因组不稳定性综合征 除其他一些毁灭性的临床表现外，PGC耗竭和不孕症 例如骨髓衰竭和癌症易感性。 FA途径是主要的DDR途径 已知可以在双链DNA断裂修复中起作用。 PGC的信号传导机制 FA患者或小鼠模型的耗竭和不育症尚不清楚。我们最近开发了 旗帜和黑凝集素标记的fancd2敲入小鼠，使高吞吐量质量 在不同小鼠器官中搜索FANCD2结合蛋白的光谱方法。在 除了DDR合作伙伴外，我们观察到了种系特异性PRMT5/PIRNA的几种蛋白质 策划抑制转座元件（TES）的途径。删除FANCD2 导致PRMT5/PIRNA因子的蛋白质水平降低，TES的大量上调， PGC耗竭，并导致fancd2-null（fancd2-ko）中的精子发生和卵子发生有缺陷 老鼠。这些初步研究表明，除了其公认的DNA修复作用外， FANCD2和FA途径在早期生殖细胞中具有体内沉默功能。我们 假设FA途径对于涉及途径层次结构的种系完整性至关重要 其中FA核心向FANCD2发出信号，然后引导Prmt5和Pirna沉默。这 该项目的目标是在FA途径和生殖细胞之间建立功能相互作用 特定的TE沉默机制在保护种系基因组中，并将fancd2定义为 生殖细胞发育过程中这种重要表观遗传学编程的关键调节剂。 该项目提出了一项旨在将主要DDR途径连接到种系的创新研究 早期生殖细胞中的基因组维持。从拟议的研究中获得的知识 不仅会提高对FA分子协作的机械理解 在TE镇压的背景下，DDR途径和PRMT5/PIRNA途径，但也导致 新的研究途径旨在针对这些相互作用的途径来发展创新 生殖疾病的治疗策略，例如不育症和先天缺陷。