The molecular and genetic bases of diverse tissue repair responses in postembryonic Drosophila

项目摘要/摘要 对于偶尔遇到恶劣环境的多细胞生物来说，组织修复是必不可少的。这个 快速修复和恢复屏障组织功能的能力对生物体的生存至关重要，而且 保守的。我的实验室率先使用果蝇幼虫来研究胚胎后表皮伤口 治愈。过去11年(其中9项由R01GM083031资助；胚胎后基因控制 果蝇的伤口愈合)我们已经研究了启动伤口闭合的信号机制， 将炎性巨噬细胞样细胞招募到伤口，这些细胞实际上协调伤口诱导的细胞 迁移，并介导了创伤诱导的表皮细胞-细胞融合这一奇怪的现象。此NIGMS R35 Mira应用程序旨在为我的实验室提供更稳定和灵活的资金，因为它继续 寻求有关组织修复反应的重要细胞和机制问题。我们的长- 学期目标是确定启动每个重要创伤反应(迁移、细胞形状)的全套基因 改变、去分化、融合、炎症)，并了解这些基因是如何发挥作用并共同作用的 精心策划一个成功的伤口修复计划。今后五年，我们的工作重点是三个关键 从我们正在进行的研究中自然出现的问题：1.表观遗传重新编程和一种 伤口边缘细胞的协调转录反应有助于伤口闭合？这个问题是从 我们观察到，一些表观遗传调节因子从伤口边缘细胞和其他细胞中清除，以及 某些转录因子是伤口闭合所直接需要的。将作出重大努力，致力于 了解伤口边缘愈合所必需的转录和表观遗传变化。2. 我们现在已经建立了一个可行的平台来识别和研究基因的功能，这些基因是 创伤诱导的细胞-细胞融合的奇怪现象。这一努力可能会带来重大的新奇见解 未被充分研究的上皮细胞-细胞融合过程是如何被控制和协调的。3.血液是如何 细胞附着在伤口上并在伤口上扩散，以发挥清除细胞碎片的功能？我们现在有了 确定了至少一个必需的信号通路(血管内皮生长因子[VEGF]信号 因为血细胞在伤口处扩散。这表明依恋和传播在基因上是可分离的。 我们已经制定了一种策略，既可以在这个过程中确定更多的参与者，也可以研究 在这一背景下，已知的在血管内皮生长因子下游发挥作用。我的实验室丰富的创意史 关于组织修复的各个方面的高影响力的出版物强烈地表明，我们将继续沿着这一脉络， 特别是如果我们正在进行的赠款编写负担减轻的话。我们的系统与该领域的其他系统相辅相成， 继续对生物体如何在细胞和组织中应对组织损伤的新的基本见解的可能性 分子/遗传水平很高。

PROJECT SUMMARY/ABSTRACT Tissue repair is essential to multicellular organisms that occasionally encounter a hostile environment. The ability to rapidly repair and restore function to barrier tissues is critical to organismal survival and is highly conserved. My laboratory pioneered the use of Drosophila larvae to study postembryonic epidermal wound healing. Over the past eleven years (9 of them funded by R01GM083031; Genetic control of postembryonic wound healing in Drosophila) we have investigated the signaling mechanisms that initiate wound closure, that recruit inflammatory macrophage-like cells to the wound, that actually orchestrate wound-induced cell migration, and that mediate the curious phenomenon of wound-induced epidermal cell-cell fusion. This NIGMS R35 MIRA application is intended to provide more stable and flexible funding to my laboratory as it continues to pursue important cellular and mechanistic questions regarding organismal tissue repair responses. Our long- term goal is to identify the full suite of genes that initiate each important wound response (migration, cell shape change, dedifferentiation, fusion, inflammation) and understand how these genes function and work together to orchestrate a successful wound repair program. Our work over the next five years will focus on three essential questions that emerge naturally from our ongoing studies: 1. How does epigenetic reprogramming and a coordinated transcriptional response in wound edge cells help facilitate closure? This question emerges from our observation that some epigenetic regulators are cleared from wound edge cells and others, along with certain transcription factors, are directly required for wound closure. A major effort will be devoted to understanding the transcriptional and epigenetic changes that are necessary for healing at the wound edge. 2. We have now set up a viable platform for identifying and studying the function of genes that are necessary for the curious phenomenon of wound-induced cell-cell fusion. This effort is likely to lead to major novel insights into how the understudied process of epithelial cell-cell fusion is controlled and orchestrated. 3. How do blood cells adhere to and spread at a wound to serve their function of clearing cellular debris? We have now identified at least one signaling pathway (Vascular Endothelial Growth Factor [VEGF] signaling) that is required for blood cell spreading at wound sites. This suggests that attachment and spreading are genetically separable events and we have developed a strategy for both identifying more players in this process and studying the function of known players that act downstream of VEGF in this context. My lab's substantial history of creative high-impact publications on diverse aspects of tissue repair suggests strongly that we will continue in this vein, especially if our ongoing grant-writing burden is reduced. Our system complements others in the field and the likelihood of continued novel basic insight into how organisms cope with tissue damage at the cellular and molecular/genetic levels is high.