Regulation of cell growth and proliferation

项目总结 该MIRA/R35应用旨在取代R01授权GM126033和GM124434。这些项目 解决细胞生长和增殖的调节，这是细胞和发育生物学的中心主题 与其中细胞生长被失调的许多人类疾病有关(例如，退行性疾病， 炎症性疾病、癌症)。我们由NIGMS支持的研究可以追溯到1994年， 目的了解在复杂的动物体内环境下，细胞增殖是如何被调节的。我们的 研究解决了基本问题：生长信号如何驱动细胞生长，细胞生长如何与 新陈代谢调节细胞周期进程，以及环境、细胞和分子相互作用如何调节 体内的生长信号活动。我们主要在果蝇中使用遗传方法，目前的重点是 肠道上皮细胞更新，但我们也抓住机会用人类细胞和 有机化合物。我们的两个项目正在测试非正统的假设，即依赖于增长的翻译 编码限制细胞周期调节因子的mRNA决定了细胞是否增殖以及增殖的速度有多快。我们有 在果蝇和人类细胞中验证了这一机制，目前正在研究上游 EGFR/RAS/ERK和Insulin/PI3K/mTOR信号转导与生长相关因子的相互作用 促进DNA复制的基因(如E2F1、CCNE2、CDC45)。第三个相关项目将扩大我们的发现 EGFR/ERK信号促进线粒体生物发生和代谢转变，激活细胞生长和 在果蝇和人类细胞中都有增殖。解释生长如何与细胞相耦合的新范式 组织可以提出新的策略和基因靶点来诊断、治疗和预防常见的 涉及细胞增殖失调的疾病。最后两个项目集中在果蝇肠道是如何 感知损害并对其作出反应。这与增殖控制有关，因为对于大多数上皮细胞来说，损伤 启动包括生长信号、干细胞激活和受调控细胞在内的再生反应 组织。上皮损伤反应也会刺激炎症反应，从而进一步与临床相关。至 为了理解这种再生反应，基因组学方法(atac-seq、Cut&Tag、RNA-seq)将被用于 确定损伤相关细胞因子/JAK/STAT信号转导的靶基因。此外，我们正在进行一项 独特的高通量功能筛选，使用肠细胞靶向RNAi来识别所有果蝇 感知肠道上皮损伤和启动再生所需的基因。基因的全面目录 用于组织损伤反应将是扩展我们对压力的理解的基础资源- 激活、炎症和再生信号。这将反过来提供新的方法：1)控制 感染期间和自身免疫性疾病中的炎症，以及：2)刺激再生以对抗 退行性疾病，并协助组织工程。

PROJECT SUMMARY This MIRA/R35 application was conceived to replace R01 grants GM126033 and GM124434. These projects address the regulation of cell growth and proliferation, a central topic in cell and developmental biology that is relevant to the many human disorders in which cell growth is dysregulated (e.g. degenerative diseases, inflammatory conditions, cancers). Our NIGMS-supported research reaches back to 1994, with the singular objective to understand how cell proliferation is regulated in vivo, in the complex context of the animal body. Our research addresses fundamental issues: how growth signaling drives cell growth, how cell growth-associated metabolism regulates cell cycle progression, and how environmental, cellular and molecular interactions regulate growth signaling activities in vivo. We primarily utilize genetic approaches in Drosophila, with a current focus on intestinal epithelial renewal, but we also seize opportunities to translate our findings using human cells and organoids. Two of our projects are testing the unorthodox hypothesis that growth-dependent translation of mRNAs encoding limiting cell cycle regulators determines whether, and how fast, cells proliferate. We have validated this mechanism in Drosophila and human cells, and are currently investigating how upstream EGFR/RAS/ERK and Insulin/PI3K/mTOR signaling interface with the growth-dependent translation of factors that promote DNA replication (e.g. E2F1, CCNE2, CDC45). A third, related project will extend our discovery that EGFR/ERK signaling promotes mitochondrial biogenesis and a metabolic shift that activates cell growth and proliferation, in both Drosophila and human cells. New paradigms explaining how growth is coupled to cell division can present novel strategies and gene targets for the diagnosis, treatment, and prevention of common diseases involving dysregulated cell proliferation. Two final projects focus on how the Drosophila intestine senses and responds to damage. This is relevant to proliferative control because, for most epithelia, damage initiates a regenerative response that comprises growth signaling, stem cell activation, and regulated cell division. Epithelial damage responses also stimulate inflammation, giving further clinical relevance. To understand this regenerative response, genomics approaches (ATAC-seq, Cut&Tag, RNA-seq) will be used to identify the target genes of damage-dependent Cytokine/Jak/Stat signaling. In addition, we are conducting a unique, high-throughput functional screen using enterocyte-targeted RNAi’s to identify all of the Drosophila genes required to sense gut epithelial damage and initiate regeneration. A comprehensive catalog of the genes used in tissue damage responses will be a foundational resource for extending our understanding of stress- activated-, inflammatory-, and regenerative signaling. This will in turn present new approaches for: 1) controlling inflammation during infections and in auto-immune diseases, and: 2) stimulating regeneration to combat degenerative disease, and to aid tissue engineering.