Understanding how cells invade through basement membrane in vivo

了解体内细胞如何侵入基底膜

• 批准号: 10631095
• 负责人: David R Sherwood
• 金额: $ 61.66万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10631095
• 关键词: Adhesions; Arthritis; Asthma; Basement membrane; Behavior; Biological Process; Caenorhabditis elegans; Cell membrane; Cell model; Cell physiology; Cells; Clinical Trials; Communication; Development; Disease; Disseminated Malignant Neoplasm; Electron Transport; Enzymes; Event; Extracellular Matrix; Genetic; Genetic Transcription; Genomic approach; Glucose; Health; Human; Immune; Immune System Diseases; Infection; Injury; Invaded; Lipids; Malignant Neoplasms; Matrix Metalloproteinase Inhibitor; Matrix Metalloproteinases; Membrane; Mission; Mitochondria; Molecular; Multiple Sclerosis; Site; Stereotyping; Stress; Stroke; Structure; Therapeutic; Tissues; Transcriptional Regulation; United States National Institutes of Health; Visual; Visualization; Work; cell type; developmental disease; experimental study; functional genomics; genetic manipulation; genome editing; healing; human disease; improved; in vitro Assay; in vivo; in vivo Model; lipid biosynthesis; migration; novel therapeutic intervention; overexpression; prevent; programs; sensor; trafficking

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.

项目概要 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。期间 发育和免疫细胞运输，特化细胞获得突破 BM 屏障的独特能力 分散、构建组织并迁移到感染和损伤部位。细胞侵袭也不当 在许多疾病期间启动，是哮喘、中风、关节炎、多发性硬化症等组织破坏的基础 硬化症和转移性癌症。因此，了解细胞如何穿越 BM 屏障至关重要 对改善人类健康具有重要意义。细胞侵袭事件通常是随机的、快速的，并且涉及动态 入侵细胞、BM 和邻近组织之间的粘附和通讯。由于这个 复杂性，不可能通过体外测定忠实地重现细胞侵袭，并且很难 对脊椎动物组织中的入侵进行可视化和基因剖析。因此，细胞的潜在机制 入侵行为仍然知之甚少。线虫中的锚细胞侵袭是体内高度定型的 细胞侵袭模型独特地结合了许多强大的实验方法，包括亚细胞 细胞-BM相互作用的可视化分析、分子活性传感器、快速基因组编辑、细胞类型特异性基因 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 研究将描述入侵细胞如何获取和使用能量来促进骨髓入侵。这部作品将揭示 指导极化葡萄糖输入的机制和专门的电子传输链的构建 丰富的线粒体提供局部 ATP 为 BM 破坏机制提供动力。此外，概述的 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 计划以及在大多数转移性癌症中过度表达的脂质产生酶如何构建一个大的、 短暂的、侵入性的突出，打开穿过BM屏障的路径。拟议的研究还将阐明如何 侵袭性细胞通过物理方式调整其侵袭程序以适应基质金属蛋白酶（MMP）的缺失 取代 BM，这将提供更有效的方法来阻止 MMP 抑制剂的入侵 迄今为止在临床试验中未能取得有效效果。最后，这项工作将确定阻止的分子机制 并修复 BM 破坏期间的质膜损伤，从而揭示可利用的机制 在入侵行为中针对目标细胞。这些综合研究涵盖细胞能量学、细胞外基质、 转录调控和膜动力学与 NIH 的使命相关，因为它们将导致 更深入地了解细胞侵袭行为的基本生物过程，从而允许 开发更好的治疗策略来调节人类疾病的侵袭。