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: 10490283
• 负责人: ROGER D KAMM
• 金额: $ 31.05万
• 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/10490283
• 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; Coping Skills; 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; 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：总结 转移定植需要循环肿瘤细胞(CTCs)克服物理应激源和动态平衡。 使成功转移成为不可能的结果的障碍。人们对转移的亚群知之甚少， 使他们能够绕过体内平衡障碍的适应，以及用来应对这些应激源的机制 要么繁殖，要么进入休眠。血管内环境对CTC来说是不适宜的，但 多条临床证据表明，与活化的血小板、纤维蛋白血栓、免疫细胞的物理相互作用 而与其他癌细胞形成的簇状会影响转移潜力。此外，其作用机制也是 微血管内的外渗是由内皮细胞相互作用、细胞骨架力量、核 变形和基质蛋白分解。长期以来，人们认识到转移的趋向性是由内在的决定的。 器官属性。我们假设二次殖民是一系列小概率事件的结果 只有一小部分CTC适应了这些应激源的事件。为了调查 停滞、外渗和定植的机制我们已经建立了体外血管网络，这些网络可以重现 微血管网络的几何形状和功能，循环中的肿瘤细胞在这里发起转移病变。 重要的是，我们能够通过控制细胞成分来精确地设计微血管环境， 细胞外成分和物理应激源，以系统区分特定扰动对 高时间和空间分辨率的癌细胞停滞、迁移和定植。在目标1中，我们创建了 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、轮回和殖民。在目标2中，我们将我们的微血管平台的功能扩展到 总结肝脏和真皮器官特有的微血管环境，以检查不同 血流和内皮屏障功能。在目标3中，我们将使用特定的分子干预来靶向肿瘤细胞 粘附性、可缩性、核变形性和基质降解以量化对血管内粘连的影响， 跨内皮细胞迁移和长期的血管外命运。在目标4中，我们将测量原子核形变和 量化转位过程中的染色质重组，并确定染色质的定量测量 重组使渗出的细胞变成休眠的表型(核心B)。总而言之，我们假设 方法性体外观察结合活体研究(项目2)和计算验证 建模(核心A)将导致对使CTC能够绕过物理 压力源。通过设计物理环境，我们将产生导致新的治疗方法的知识 阻止或逆转应对表型的机会。