The role of ESCRTs in signaling within the testis stem cell niche

ESCRT 在睾丸干细胞生态位内信号传导中的作用

• 批准号: 10464094
• 负责人: Mara Ruth Grace
• 金额: $ 4.68万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10464094
• 关键词: Ablation; Affect; Aging; Animal Model; Back; Cell Count; Cell Lineage; Cell Maintenance; Cell Size; Cells; Cellular Morphology; Communication; Complement; Complex; Confocal Microscopy; Core Facility; Cyst; Data; Data Set; Daughter; Development; Developmental Process; Drosophila genus; Drosophila melanogaster; Educational process of instructing; Educational workshop; Electron Microscopy; Endocytosis; Ensure; Equilibrium; Flow Cytometry; Fluorescence; Foundations; Gametogenesis; Gene Expression; Genes; Genetic; Genomics; Germ; Grant; Growth; Homeostasis; Hypertrophy; Immune Sera; Injury; Intestines; Longevity; Maintenance; Malignant Neoplasms; Manuscripts; Measurement; Mediating; Mentors; Mesenchymal; Microscopy; Morphology; Mosaicism; Natural regeneration; Oncology; Organism; Pathway Analysis; Pathway interactions; Phenotype; Ploidies; Polyploidy; Process; RNA Interference; Regenerative Medicine; Reproduction; Research; Research Training; Resources; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Spermatogenesis; Surveys; System; Testing; Testis; Tissues; Transgenic Organisms; Tumor Suppressor Genes; Work; Writing; adult stem cell; cell growth; experience; experimental study; fly; follow-up; gain of function; insight; knock-down; medical schools; member; mutant; novel; receptor; self-renewal; skills; stem; stem cell biology; stem cell niche; stem cell population; stem cells; symposium; teaching assistant; tissue regeneration; tissue repair; tool; transcriptomics; tumorigenesis; undergraduate student

PROJECT SUMMARY/ABSTRACT Adult stem cells are crucial for regeneration, tissue repair after injury, and developmental processes such as spermatogenesis. These cells ensure both tissue growth and renewal by dividing asymmetrically, producing one daughter stem cell and one daughter that differentiates 1,2. Stem cells exist in a dynamic microenvironment termed the niche that provides signals to ensure the maintenance and self-renewal of the adult stem cell population1–5. An appreciation of the dynamic communication between stem cells and their niche is vital to understand processes such as reproduction, oncogenesis, aging, development, and regeneration. Much is known about signals sent from niche cells and received by stem cells3,6–13; however, little is known about signaling in the opposite direction: from stem cells back to their niche. Here, I use the testis stem cell niche of Drosophila melanogaster to explore signaling from stem cells back to their niche as well as the role of endocytic tumor suppressor genes in this signaling. Endocytosis regulates a myriad of signaling pathways14–18. My preliminary data indicate that knockdown of an endocytic gene in somatic stem cells greatly increases the size of adjacent niche cells and, consequently, the niche itself. Since the way that endocytic genes non-autonomously regulate niche size is unknown, but may apply widely to other niches. In Aim 1, I further characterize this phenotype by assessing the role of endocytosis in mediating signaling within the Drosophila testis stem cell niche. I will specifically knock down genes regulating endocytosis from cyst stem cells and characterize the resultant changes in neighboring niche cells, focusing on niche cell size, morphology, and ploidy. I will also examine the activity of a small set of well-characterized signaling pathways involved in cell growth in wild-type and endocytic mutant testes. I will follow up with genetic loss and gain-of- funditon analysis for pathways with changes such as the JAK-STAT pathway (preliminary data). In Aim 2, I more broadly investigate signaling from other cells to the niche cells by performing a screen of candidate niche cell signaling pathway receptors identified in our recent single-cell transcriptomic datasets. This will greatly expand our understanding of the signaling pathways requred for niche cell fate, function, size and/or quiescence. Overall, these proposed experiments will further illuminate cell signaling dynamics within the Drosophila testis stem cell niche, with implications for niches in other tissues and organisms. I will conduct this work in the lab of Dr. Erika Matunis at the Johns Hopkins School of Medicine, where I have access to over 50 departmental and core facilities and resources including Drosophila media, flow cytometry, microscopy, genetics and genomics, and a supportive mentoring team. I will supplement my research training by strengthening my quantitative skills through workshops and courses, and my communication skills through grant and manuscript writing and presentations at conferences. Furthermore, I will gain teaching experience by serving as a teaching assistant and mentoring undergraduates in the lab.

项目总结/摘要 成体干细胞对于再生、损伤后的组织修复和发育过程至关重要， 精子发生这些细胞通过不对称分裂，产生 一个子干细胞和一个分化的子细胞1，2.干细胞存在于一个动态的 微环境称为生态位，提供信号，以确保维护和自我更新的 成体干细胞群1 -5.干细胞和它们之间的动态通讯的欣赏 生态位对于理解生殖、肿瘤发生、衰老、发育和 再生关于从小生境细胞发出并由干细胞接收的信号，我们知道得很多3，6-13;然而， 对于相反方向的信号传递知之甚少：从干细胞回到它们的小生境。在这里，我使用 黑腹果蝇睾丸干细胞生态位，探索从干细胞返回其生态位的信号传导， 以及内吞肿瘤抑制基因在这种信号传导中的作用。内吞作用调节了无数的 信号通路14 -18。我的初步数据表明，在体细胞茎中敲除内吞基因， 细胞极大地增加了相邻的小生境细胞的大小，从而增加了小生境本身的大小。因为 内吞基因非自主调节生态位大小是未知的，但可能广泛适用于其他生态位。在 目的1，我通过评估内吞作用在介导细胞内信号传导中的作用，进一步表征这种表型。 果蝇睾丸干细胞龛我会专门敲除调控胞吞作用的基因 干细胞，并表征相邻小生境细胞中的结果变化，重点是小生境细胞大小， 形态学和倍性。我还将研究一小部分特征明确的信号通路的活性 参与野生型和内吞突变体睾丸的细胞生长。我会跟进基因缺失和获得- 对JAK-STAT通路等发生变化的通路进行基础分析（初步数据）。在目标2中，我 通过进行候选小生境的筛选来更广泛地研究从其它细胞到小生境细胞的信号传导 在我们最近的单细胞转录组数据集中鉴定的细胞信号通路受体。这将大大 扩大我们对小生境细胞命运、功能、大小和/或 安静总的来说，这些拟议的实验将进一步阐明细胞内的信号转导动力学。 果蝇睾丸干细胞生态位：对其他组织和生物生态位的启示。 我将在约翰霍普金斯医学院的Erika Matunis博士的实验室进行这项工作， 使用50多个部门和核心设施和资源，包括果蝇培养基，流式细胞术， 显微镜，遗传学和基因组学，以及一个支持性的指导团队。我会补充我的研究训练 通过研讨会和课程加强我的定量技能，通过 资助和手稿写作以及在会议上的演讲。此外，我将获得教学经验 通过在实验室担任助教和指导本科生。