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.

项目概要/摘要 成体干细胞对于再生、损伤后的组织修复以及发育过程（例如 精子发生。这些细胞通过不对称分裂确保组织生长和更新，产生 一个子代干细胞和一个分化1,2的子代细胞。干细胞以动态状态存在 微环境被称为生态位，它提供信号以确保生物体的维护和自我更新 成体干细胞群1-5。了解干细胞及其之间的动态通讯 生态位对于理解生殖、肿瘤发生、衰老、发育和生长等过程至关重要。 再生。关于从生态位细胞发送并由干细胞接收的信号已经有很多了解3,6-13；然而， 人们对相反方向的信号传导知之甚少：从干细胞回到它们的生态位。在这里，我使用 黑腹果蝇睾丸干细胞生态位探索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此信号传导中的作用。内吞作用调节多种 信号通路14-18。我的初步数据表明体细胞干中内吞基因的敲低 细胞大大增加了相邻生态位细胞的大小，从而增加了生态位本身的大小。既然这样 内吞基因非自主调节生态位大小尚不清楚，但可能广泛应用于其他生态位。在 目标 1，我通过评估内吞作用在介导内吞信号中的作用来进一步表征这种表型 果蝇睾丸干细胞生态位。我将专门敲除囊肿中调节内吞作用的基因 干细胞并表征邻近生态位细胞的变化，重点关注生态位细胞的大小， 形态学和倍性。我还将检查一小组特征明确的信号通路的活动 参与野生型和内吞突变型睾丸的细胞生长。我将跟进遗传损失和获得- 对 JAK-STAT 通路等变化的通路进行基础分析（初步数据）。在目标 2 中，我 通过筛选候选生态位，更广泛地研究从其他细胞到生态位细胞的信号传导 我们最近的单细胞转录组数据集中发现了细胞信号通路受体。这将大大 扩大我们对微环境细胞命运、功能、大小和/或所需信号通路的理解 静止。总体而言，这些拟议的实验将进一步阐明细胞信号传导动态 果蝇睾丸干细胞生态位，对其他组织和生物体的生态位具有影响。 我将在约翰·霍普金斯大学医学院的 Erika Matunis 博士的实验室中进行这项工作，我在那里 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，以及支持性的指导团队。我将补充我的研究训练 通过研讨会和课程加强我的定量技能，并通过 资助和手稿写作以及在会议上的演讲。此外，我还将获得教学经验 在实验室担任助教并指导本科生。