Molecular Mechanisms of Leydig Cell Regeneration

间质细胞再生的分子机制

• 批准号: 10454800
• 负责人: Jasmin LaKia Jeffery
• 金额: $ 4.68万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10454800
• 关键词: Ablation; Adult; Age; Aging; Alkanesulfonates; Androgens; Automobile Driving; Biology; Candidate Disease Gene; Cell Cycle; Cell Death; Cell Proliferation; Cells; Characteristics; Chemicals; Cre driver; Cytotoxic agent; Data; Development; Ethane; Gene Expression Profile; Gene Expression Profiling; Genes; Genetic; Germ Cells; Goals; In Vitro; Injury; Interphase Cell; Male Infertility; Malignant Neoplasms; Modeling; Molecular; Monitor; Mus; Natural regeneration; Organ; Organism; Pathway interactions; Personal Satisfaction; Pharmacology; Physiological; Population; Process; Rattus; Regenerative research; Role; Signal Pathway; Signal Transduction; Somatic Cell; Source; Supporting Cell; System; Testing; Testis; Tissue Model; Tissues; Work; candidate marker; cell regeneration; cell type; differential expression; extracellular; improved; in vivo regeneration; inhibitor; injured; interest; leydig interstitial cell; male; male fertility; male health; men; mouse genetics; novel; precursor cell; progenitor; regeneration model; regenerative; regenerative cell; repaired; response; response to injury; self-renewal; single-cell RNA sequencing; small molecule; sperm cell; stem cells; tissue regeneration; tool; transcriptome; transcriptome sequencing; transcriptomics

Cells lost to normal wear and tear in many adult tissues are replaced with the activity of stem cells. In response to injury, however, cells can use entirely different strategies to repair and regenerate tissues. Injured tissues can be regenerated through the activation of stem cells or from progenitor cells that, unlike stem cells, are incapable of asymmetric division and self-renewal. The dramatic cellular changes that occur during regeneration are thought to arise from altered signaling in the tissue microenvironment, or niche, but the mechanisms regulating regeneration are largely unknown. A classical model of regeneration after injury is the adult rat testis. In the testis, somatic cells create a microenvironment that contributes to the germ cell niche. Critical somatic support cells include Leydig cells, the major androgen producing cells in males, which are required for male health and fertility. Although Leydig cells are a quiescent cell type, and not thought to divide under normal conditions, they are chemically ablated by ethane dimethane sulfonate (EDS). Several weeks after chemical ablation, new Leydig cells repopulate the testis. The mechanism of Leydig cell regeneration and the progenitors from which they arise remain poorly understood, partially due to the lack of genetic tools available in rats. In vitro work has hinted at candidate markers that may identify progenitors of Leydig cell regeneration. The goal of this project is to determine the cellular mechanism and molecular mechanism of Leydig cell regeneration. Mouse genetics make it possible to interrogate the cellular mechanism of regeneration by use of an inducible genetic lineage tracing system. In preliminary data, a subset of somatic testis interstitial cells trace into new Leydig cells after ablation in mice. We will continue using this model to determine the both identity of the progenitor cells and whether this cell type is self-renewing. We will also determine the molecular mechanisms driving regeneration with the use of sc-RNA sequencing before, during, and after regeneration. Candidate genes implicated in signaling and Leydig cell development that are differentially expressed in regenerating cells will be tested for functional roles during regeneration using small molecule pathway inhibitors and available genetic tools. While small molecules are an accessible way to gain crude information about whether a signaling pathway is involved, sc-RNA sequencing will illuminate exactly which cells produce signals that initiate the progenitor response. Determining the cellular and molecular mechanisms of Leydig cell regeneration will significantly contribute to the understanding of adult tissue regeneration in vivo with broader implications for male fertility and health.

许多成年组织中失去正常磨损的细胞被干细胞的活性取代。作为回应 然而，要受伤，细胞可以使用完全不同的策略来修复和再生组织。受伤的组织 可以通过干细胞的激活或与干细胞不同的祖细胞的激活来再生 无效分裂和自我更新。在 人们认为再生是由组织微环境或利基市场中的信号改变引起的，但是 调节再生的机制在很大程度上未知。受伤后的经典再生模型是 成年大鼠睾丸。在睾丸中，体细胞会产生一个微环境，从而有助于生殖细胞生态位。 关键的体细胞支撑细胞包括Leydig细胞，雄性中主要雄激素产生细胞， 男性健康和生育需要。尽管leydig细胞是一种静止的细胞类型，而不是认为分裂 在正常条件下，它们在化学上被乙烷二甲烷磺酸盐（EDS）烧蚀。几个星期 化学消融后，新的Leydig细胞重新填充了睾丸。 Leydig细胞再生的机制和 它们出现的祖细胞仍然很糟糕，部分原因是缺乏遗传工具 可在老鼠中使用。体外工作暗示了候选标记，这些标记可能鉴定出Leydig细胞的祖细胞 再生。该项目的目的是确定细胞机制和分子机制 Leydig细胞再生。小鼠遗传学使审问细胞机制成为可能 通过使用诱导的遗传谱系追踪系统再生。在初步数据中，躯体的一部分 睾丸间质细胞在小鼠消融后将其追踪到新的Leydig细胞中。我们将继续使用此模型 确定祖细胞的同一身份以及该细胞类型是否自我更新。我们也会 通过使用SC-RNA测序在，在 并在再生之后。与信号传导和leydig细胞发育有关的候选基因 使用小 分子途径抑制剂和可用的遗传工具。而小分子是一种获取的方法 关于是否涉及信号通路的粗略信息，SC-RNA测序将精确照亮 哪些细胞产生启动祖细胞反应的信号。确定细胞和分子 Leydig细胞再生的机制将显着有助于理解成人组织 体内再生，对男性生育和健康具有更广泛的影响。