Elucidating function of the novel PINX1-related protein, Chigno, in reproductive stem cells and development

阐明新型 PINX1 相关蛋白 Chigno 在生殖干细胞和发育中的功能

• 批准号: 10730857
• 负责人: MATTHEW J WAWERSIK
• 金额: $ 43.35万
• 依托单位: COLLEGE OF WILLIAM AND MARY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10730857
• 关键词: Adult; Apical; Authorship; Basic Science; Behavior; Binding; Binding Proteins; Cell Differentiation process; Cell Maintenance; Cell physiology; Cells; Cessation of life; Chronology; Communication; Contraceptive methods; Cyst; Data; Daughter; Development; Drosophila genus; Drosophila melanogaster; Educational process of instructing; Embryo; Ensure; Environment; Epithelial Cells; Equilibrium; Fertility; Fostering; Future; Gene Expression; Genes; Germ Cells; Germ cell tumor; Goals; Gonadal structure; Grant; Homeostasis; Human; Human Development; Infertility; Link; Maintenance; Malignant Neoplasms; Mammals; Maps; Methodology; Molecular; Morphogenesis; Organ; Organism; Ovary; Pattern; Process; Production; Proteins; Records; Regulation; Repression; Research; Research Personnel; Research Training; Resources; Role; Scholarship; Students; System; Testing; Testis; Tissues; Training; Tumor Suppressor Proteins; Work; career; cell behavior; developmental disease; embryo tissue; experimental study; fly; gene regulatory network; germline stem cells; human disease; in vivo; knock-down; neuronal cell body; novel; prevent; programs; protein function; protein protein interaction; reproductive; senescence; stem cell niche; stem cell self renewal; stem cells; student participation; student training; success; symposium; training opportunity; transcription factor; tumor; undergraduate student

PROJECT SUMMARY: The germline stem cell (GSC) niche functions to ensure continuous gamete production in organisms that reproduce sexually. Dysregulation of stem cells has also been linked to developmental disorders, cancer, infertility, and senescence. In flies and mammals alike, communication between GSCs and the surrounding soma regulates the balance between stem cell maintenance and differentiation. However, new and unexplored mechanisms regulating these processes are still being discovered - necessitating further enquiry. This proposal strives to elucidate the function of a novel PINX1-related protein, CG11180/Childless Gambino (Chigno), in regulating reproductive stem cell behavior and development in the fruit fly, Drosophila melanogaster. Chigno is a novel G-patch domain-containing protein in flies that is related to the mammalian tumor suppressor, PINX1. We previously identified Chigno as a binding partner of Chronologically inappropriate morphogenesis (Chinmo), a key regulator of somatic stem cell behavior in the adult Drosophila testis. Preliminary data collected by undergraduate and master’s student researchers in our lab show that Chigno is expressed in larval and adult testes, as well as Drosophila ovaries and early embryos. In adult testes, Chigno is observed specifically at the testis apex, which houses the germline stem cell (GSC) niche. Here, it is detected both in germ cells as well as somatic cyst stem cells (CySCs) and their progeny which regulate germline behavior. We also find that Chigno knockdown in adult CySCs and early cyst cells results in CySC over-production, formation of germ cell tumors, and an expansion of epithelial cells at the testis apex that may be linked to altered CySC identity. Furthermore, somatic Chigno depletion causes both testis and ovaries to become infertile. Experiments in this proposal are focused on understanding the role of Chigno in regulating testis stem cell self-renewal, differentiation, and cell fate maintenance (Aim 1), understanding Chigno protein function at the molecular level (Aim 2), and assessing whether Chigno functions in tissues other than the testis, including Drosophila ovaries and embryonic tissues (Aim 3). Given Chigno’s impact on fertility and testes stem cell behavior, as well as the diverse functions of G-patch/PINX1-related proteins in a range of cellular processes, the proposed studies into Chigno function hold great promise for elucidating novel mechanisms controlling fertility, stem cell behavior, and development in a range of organisms.

项目概要： 生殖系干细胞（GSC）小生境的功能是确保生物体中持续的配子产生， 有性繁殖。干细胞的失调也与发育障碍、癌症、 不育和衰老在果蝇和哺乳动物中，GSC和周围环境之间的交流 索马调节干细胞维持和分化之间的平衡。然而，新的和 目前仍在发现尚未探索的管理这些进程的机制，因此需要进一步调查。 该提案致力于阐明一种新型PINX 1相关蛋白CG 11180/Childless Gambino的功能 （Chigno），在调节生殖干细胞的行为和发展，果蝇 黑腹菌 Chigno是一种新的含有G-补丁结构域的蛋白质，与哺乳动物肿瘤有关 抑制基因PINX 1。我们以前确定Chigno是一个时间顺序不适当的结合伙伴， 形态发生（Chinmo），成体果蝇睾丸中体干细胞行为的关键调节因子。 我们实验室的本科生和硕士生研究人员收集的初步数据显示，Chigno是 在幼虫和成虫睾丸以及果蝇卵巢和早期胚胎中表达。在成年睾丸中，Chigno 特别是在睾丸顶端观察到，睾丸顶端容纳生殖系干细胞（GSC）小生境。在这里 在生殖细胞以及体细胞囊肿干细胞（CySC）及其后代中均检测到，这些细胞调节 生殖细胞行为我们还发现，在成年CySC和早期囊肿细胞中，Chigno敲低导致CySC 过度生产，生殖细胞肿瘤的形成，以及睾丸顶端上皮细胞的扩张， 与改变的CySC身份有关。此外，体细胞Chigno耗竭导致睾丸和卵巢 变得不育。 本实验的重点是了解Chigno在睾丸干细胞调控中的作用 细胞自我更新，分化和细胞命运维持（目标1），了解Chigno蛋白的功能， 分子水平（目标2），并评估Chigno是否在睾丸以外的组织中发挥作用，包括 果蝇卵巢和胚胎组织（目的3）。鉴于Chigno对生育能力和睾丸干细胞的影响， 行为，以及G-patch/PINX 1相关蛋白在一系列细胞过程中的不同功能， 对Chigno函数的拟议研究对于阐明新的控制机制具有很大的希望， 生育力、干细胞行为和一系列生物体的发育。