Understanding how cells invade through basement membrane in vivo

项目总结 基底膜(BM)是一种致密的片状细胞外基质，包围着大多数组织。在.期间 发育和免疫细胞运输，特化细胞获得独特的能力，突破骨髓障碍 分散，构建组织，并迁移到感染和受伤的地方。细胞入侵也是不适当的 起始于多种疾病，是哮喘、中风、关节炎、多发性硬化症的组织破坏的基础 硬化症和转移性癌症。因此，了解细胞如何穿越BM屏障具有基础性意义 在改善人类健康方面的重要性。细胞入侵事件通常是随机的、快速的，并且涉及动态 侵袭细胞、骨髓和邻近组织之间的黏附和通讯。由于这一点 复杂性，不可能用体外实验真实地概括细胞的侵袭，而且一直很困难。 对脊椎动物组织的入侵进行可视化和基因解剖。因此，细胞的潜在机制 侵入性行为仍然鲜为人知。线虫体内锚定细胞入侵是一种高度刻板的现象 细胞入侵模型，独一无二地结合了许多强大的实验方法，包括亚细胞 细胞-骨髓相互作用的可视化分析、分子活性传感器、快速基因组编辑、细胞类型特定基因 操作，以及强大的正向遗传和功能基因组方法。利用这些优势，这 研究将表征入侵细胞如何获得和使用能量来推动BM的入侵。这项工作将揭示 引导极化葡萄糖输入的机制和专门化电子传递链的构建 丰富的线粒体，提供局部的ATP，为BM破坏机制提供动力。此外，概述了 实验将确定脂质生物合成如何整合到保守的细胞侵袭转录中 程序以及在大多数转移性癌症中过度表达的脂质产生酶是如何建立一个巨大的， 短暂的侵入性突起，打开穿过BM屏障的路径。这项拟议的研究还将阐明 侵袭细胞通过物理方式使其侵袭程序适应基质金属蛋白酶(MMPs)的缺失 取代骨髓，这将提供更有效的方法来阻止基质金属蛋白酶抑制剂的侵袭 到目前为止，该药在临床试验中未能奏效。最后，这项工作将确定阻止 并修复BM破坏过程中的质膜损伤，从而揭示了可以利用的机制 在入侵行为中以细胞为目标。这些综合研究涵盖了细胞能量学、细胞外基质、 转录调控和膜动力学与NIH的任务相关，因为它们将导致 更深入地理解细胞侵袭行为的基本生物学过程，从而使 开发更好的治疗策略来调节人类疾病的侵袭性。

PROJECT SUMMARY Basement membrane (BM) is a dense, sheet-like extracellular matrix that surrounds most tissues. During development and immune cell trafficking, specialized cells acquire the unique ability to breach BM barriers to disperse, construct tissues, and migrate to sites of infection and injury. Cell invasion is also inappropriately initiated during numerous diseases and underlies tissue destruction in asthma, stroke, arthritis, multiple sclerosis, and metastatic cancer. Understanding how cells traverse BM barriers is thus of fundamental importance in improving human health. Cell invasion events are often stochastic, rapid, and involve dynamic adhesions and communication between the invading cell, the BM, and the neighboring tissues. Owing to this complexity, it is not possible to faithfully recapitulate cell invasion with in vitro assays, and it has been difficult to visualize and genetically dissect invasion in vertebrate tissues. As a result, the mechanisms underlying cell invasive behavior remain poorly understood. Anchor cell invasion in C. elegans is a highly stereotyped in vivo model of cell invasion that uniquely combines many powerful experimental approaches including subcellular visual analysis of cell-BM interactions, molecular activity sensors, rapid genome editing, cell-type specific gene manipulation, and powerful forward genetic and functional genomic approaches. Using these strengths, this study will characterize how invading cells acquire and use energy to fuel BM invasion. This work will reveal mechanisms that direct polarized glucose import and the construction of specialized electron transport chain enriched mitochondria that provide localized ATP to power the BM breaching machinery. Further, the outlined experiments will determine how lipid biosynthesis is integrated into a conserved cell invasion transcriptional program and how lipid producing enzymes, which are overexpressed in most metastatic cancers, build a large, transient, invasive protrusion that opens paths through BM barriers. The proposed study will also elucidate how invasive cells adapt their invasion program to the absence of matrix metalloproteinases (MMPs) by physically displacing the BM, which will inform more effective approaches to block invasion with MMP inhibitors that have thus far failed to be effective in clinical trials. Finally, this work will identify molecular mechanisms that prevent and heal plasma membrane damage during BM breaching, thus revealing mechanisms that could be exploited to target cells in the act of invading. These integrative studies spanning cellular energetics, extracellular matrix, transcriptional regulation, and membrane dynamics are relevant to the NIH’s mission as they will lead to a deeper understanding of the fundamental biological process of cell invasive behavior, thus allowing for the development of better therapeutic strategies to modulate invasion in human disease.