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: 10404047
• 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/10404047
• 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; 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; 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.

项目摘要 地下膜（BM）是大多数组织周围的密集的，薄薄的细胞外基质。期间 开发和免疫细胞运输，专门细胞获得了违反BM屏障的独特能力 分散，构建组织并迁移到感染和受伤部位。细胞入侵也不当 在哮喘，中风，关节炎，多个疾病中启动并构成组织破坏的基础 硬化症和转移性癌症。因此了解细胞如何遍历BM屏障的基本障碍 在改善人类健康方面的重要性。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和相邻组织之间的粘附和通信。因此 复杂性，不可能通过体外测定忠实地概括细胞入侵，这是困难的 可视化和遗传剖析脊椎动物组织中的侵袭。结果，细胞的基础机制 侵入性行为仍然知之甚少。秀丽隐杆线虫中的锚细胞入侵是一种高度定型的体内观念 唯一结合了许多强大的实验方法，包括亚细胞的模型 细胞-BM相互作用，分子活性传感器，快速基因组编辑，细胞类型特异性基因的视觉分析 操纵和强大的前瞻性遗传和功能基因组方法。利用这些优势，这个 研究将表征入侵细胞如何获取和利用能量为BM侵袭。这项工作将揭示 直接极化葡萄糖进口并构建专业电子传输链的机制 富集的线粒体可为局部ATP提供为BM违规机械提供动力。此外，概述了 实验将确定如何将脂质生物合成整合到配置的细胞转录中 程序以及如何在大多数转移性癌症中过表达的脂质产生酶如何构建一个大型的 瞬态，侵入性蛋白质通过BM屏障打开路径。拟议的研究还将阐明 侵入性细胞将其入侵程序适应不存在基质金属蛋白酶（MMP） 取代BM，该BM将告知更有效的方法，以阻止具有的MMP抑制剂的入侵 这远未在临床试验中有效。最后，这项工作将确定可以预防的分子机制 并治愈BM破裂期间的质膜损伤，从而揭示了可以利用的机制 在入侵行为中靶向细胞。这些综合研究涵盖细胞能量学，细胞外基质， 转录调节和膜动态与NIH的使命有关，因为它们将导致 对细胞浸润行为的基本生物学过程的更深入了解，从而允许 制定更好的治疗策略来调节人类疾病的入侵。