Regulation of cell growth and proliferation

项目摘要 本MIRA/R35申请旨在取代R01补助金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课题， 与其中细胞生长失调的许多人类疾病（例如退行性疾病， 炎性病症、癌症）。我们的NIGMS支持的研究可以追溯到1994年， 目的了解在动物体内复杂的环境中，细胞增殖是如何被调节的。我们 研究解决了基本问题：生长信号如何驱动细胞生长，细胞生长相关的 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 体内生长信号传导活性。我们主要在果蝇中利用遗传方法，目前的重点是 肠上皮更新，但我们也抓住机会利用人类细胞转化我们的发现， 类器官我们的两个项目正在测试一个非正统的假设，即生长依赖的翻译， 编码限制细胞周期调节因子的mRNA决定细胞是否增殖以及增殖的速度。我们有 在果蝇和人类细胞中验证了这种机制，目前正在研究上游如何 EGFR/RAS/ERK和胰岛素/PI3K/mTOR信号传导与生长依赖性因子翻译的界面 促进DNA复制（例如E2F1、CCNE2、CDC 45）。第三个相关项目将扩展我们的发现， EGFR/ERK信号转导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的增殖。解释生长如何与细胞结合的新范式 分裂可以提出新的战略和基因靶点的诊断，治疗和预防常见的 涉及细胞增殖失调的疾病。最后两个项目聚焦于果蝇的肠道 感知并响应损伤。这与增殖控制有关，因为对于大多数上皮细胞， 启动再生反应，包括生长信号传导、干细胞活化和调节细胞增殖。 师.上皮损伤反应还刺激炎症，从而提供进一步的临床相关性。到 了解这种再生反应，基因组学方法（ATAC-seq，Cut & Tag，RNA-seq）将用于 鉴定损伤依赖性细胞因子/Jak/Stat信号转导的靶基因。此外，我们正在进行一项 独特的，高通量的功能筛选，使用肠细胞靶向RNAi的，以确定所有的果蝇 这些基因是感知肠上皮损伤并启动再生所必需的。基因的综合目录 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- 激活的、炎症的和再生的信号传导。这将反过来提出新的方法：1）控制 感染和自身免疫性疾病期间的炎症，以及：2）刺激再生，以对抗 退行性疾病，并帮助组织工程。

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.