Project1: The role of intravascular pressure and shear stress on tumor cell arrest, survival and proliferation in the microvascular niche

项目1：血管内压力和剪切应力对微血管微环境中肿瘤细胞停滞、存活和增殖的作用

• 批准号: 10271567
• 负责人: ROGER D KAMM
• 金额: $ 31.69万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-17 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10271567
• 关键词: Actomyosin; Adhesions; Animal Model; Batimastat; Blood Circulation; Blood Platelets; Blood Vessels; Blood flow; Breast Cancer Cell; Breast cancer metastasis; CD44 gene; Cell Adhesion; Cell Survival; Cells; Cessation of life; Chromatin; Chromatin Structure; Clinical; Coagulation Process; Complement; Computer Models; Dermis; Endothelial Cells; Endothelium; Engineering; Environment; Event; Exhibits; Exposure to; Extravasation; Fibrin; Generations; Genetic Transcription; Geometry; Hematology; Human; Immune; In Vitro; Individual; Intervention; Knowledge; Lamin Type A; Lead; Lesion; Liver; Matrix Metalloproteinases; Measures; Mechanical Stress; Mechanics; Mediating; Methods; Microscopic; Modeling; Molecular; Morphology; Neoplasm Circulating Cells; Neoplasm Metastasis; Nuclear; Organ; Organ Model; Outcome; Peripheral; Phenotype; Physical environment; Population; Probability; Process; Proliferating; Property; Proteolysis; Resolution; Rho-associated kinase; Role; Signal Pathway; Source; Stress; Stromal Cells; Study models; System; Techniques; Testing; Therapeutic; Thrombus; Tissues; Tropism; base; cancer cell; coping; coping mechanism; design; druggable target; experience; experimental study; extracellular; hemodynamics; high resolution imaging; human disease; inhibitor/antagonist; insight; kinase inhibitor; knock-down; migration; monolayer; neoplastic cell; novel therapeutic intervention; novel therapeutics; overexpression; pressure; prevent; shear stress; single-cell RNA sequencing; stressor; transcriptomics; triple-negative invasive breast carcinoma

Project 1: SUMMARY Metastatic colonization requires that circulating tumor cells (CTCs) overcome the physical stressors and homeostatic barriers that make successful metastasis an unlikely outcome. Very little is known about metastatic subpopulations, the adaptations that allow them to circumvent homeostatic barriers, and the mechanisms used to cope with these stressors and either proliferate or enter into dormancy. The intravascular environment is known to be inhospitable to CTCs, yet several lines of clinical evidence indicate that physical interactions with activated platelets, fibrin thrombi, immune cells and the formation of clusters with other cancer cells influences metastatic potential. Furthermore, the mechanism of extravasation within the microvasculature is mediated by endothelial interactions, cytoskeletal forces, nuclear deformations, and matrix proteolysis. It has long been recognized that metastatic tropism is determined by intrinsic organ properties. We hypothesize that secondary colonization is the culmination of a sequence of low probability events for which only a small subpopulation of CTCs has adapted to cope with these stressors. To investigate the mechanisms of arrest, extravasation, and colonization we have developed in vitro vascular networks that recapitulate the geometry and function of the microvascular networks where circulating tumor cells initiate metastatic lesions. Importantly, we are able to precisely engineer the microvascular environment by controlling cellular constituents, extracellular components, and the physical stressors to systematically distinguish the effect of specific perturbations on cancer cell arrest, transmigration, and colonization with high temporal and spatial resolution. In Aim 1, we create cancer cell thrombi and clusters to determine the effect of interactions with platelets, fibrin, and cancer cells on the arrest, transmigration, and colonization. In Aim 2, we extend the capabilities of our microvascular platforms to recapitulate the organ-specific microvascular environments of liver and dermis to examine combined effects of different flow and endothelial barrier function. In Aim 3, we will use specific molecular interventions to target tumor cell adhesion, contractility, nuclear deformability, and matrix degradation to quantify the effect on intravascular adhesion, transendothelial migration, and long-term extravascular fate. In Aim 4, we will measure nuclear deformation and quantify chromatin reorganization during transmigration and determine if quantitative measures of chromatin reorganization fates extravasated cells to a dormant phenotype (Core B). Taken together, we hypothesize that methodical in vitro observation combined with and validated by intravital studies (Project 2) and computational modeling (Core A) will lead to new insights regarding the specific mechanisms that enable CTCs to circumvent physical stressors. By engineering the physical environment, we will generate the knowledge leading to novel therapeutic opportunities to block or reverse the coping phenotype.

项目1：摘要 转移性定殖要求循环肿瘤细胞（CTC）克服身体压力和稳态 使成功转移成为不太可能结果的障碍。关于转移性亚群的知之甚少 适应使它们可以规避稳态屏障的适应，以及用于应对这些压力源的机制 并且要么扩散或处于休眠状态。众所周知，血管内环境对CTC是不愉快的 几条临床证据表明，与活化血小板，纤维蛋白血栓，免疫细胞的物理相互作用 与其他癌细胞的形成簇会影响转移性潜力。此外，机制 微脉管系统内的渗出是由内皮相互作用，细胞骨架力，核介导的 变形和基质蛋白水解。长期以来人们已经认识到转移性向潮流取决于固有的 器官特性。我们假设二次定植是一系列低概率的顶点 仅CTC的小亚群的事件才适应应对这些压力源。调查 逮捕，渗出和定殖的机制我们已经开发了体外血管网络，这些网络概括 循环肿瘤细胞引发转移性病变的微血管网络的几何形状和功能。 重要的是，我们能够通过控制细胞成分，可以精确地设计微血管环境。 细胞外成分以及物理压力源以系统区分特定扰动对 癌细胞停滞，移民和定植，具有高时空和空间分辨率。在AIM 1中，我们创建 癌细胞血栓和簇，以确定与血小板，纤维蛋白和癌细胞相互作用对 逮捕，移民和殖民化。在AIM 2中，我们将微血管平台的功能扩展到 概括肝脏和真皮的器官特异性微血管环境，以检查不同的综合作用 流量和内皮屏障功能。在AIM 3中，我们将使用特定的分子干预来靶向肿瘤细胞 粘附，收缩力，核可变形性和基质降解，以量化对血管内粘附的影响， 跨内皮迁移和长期血管外命运。在AIM 4中，我们将衡量核变形和 量化移民期间染色质重组，并确定染色质的定量测量 重组的命运使细胞外部表型（核心B）脱颖而出。综上所述，我们假设 有条理的体外观察结合并通过插入性研究（项目2）和计算验证 建模（核心A）将导致有关使CTC规避物理的特定机制的新见解 压力源。通过工程化物理环境，我们将产生知识，从而导致新型治疗 阻止或扭转应对表型的机会。