Fibronectin and its role in tumor stiffness and vascularization

纤连蛋白及其在肿瘤硬度和血管化中的作用

• 批准号: 8176810
• 负责人: Claudia Fischbach
• 金额: $ 16.37万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8176810
• 关键词: 3-Dimensional; Adipose tissue; Atherosclerosis; Biochemical; Breast; Breast Cancer Cell; Breast Cancer Treatment; Cells; Characteristics; Collagen; Conditioned Culture Media; Cues; Deposition; Elasticity; Endothelial Cells; Event; Extracellular Matrix; Fiber; Fibronectins; Fluorescence Resonance Energy Transfer; Goals; Growth; Image; In Vitro; Lead; Mammary Neoplasms; Measures; Mechanics; Mediating; Modeling; Molecular; Molecular Conformation; Organogenesis; Paracrine Communication; Pathogenesis; Pathologic Processes; Phenotype; Physiological; Physiological Processes; Play; Process; Property; Research; Role; Signal Transduction; Signaling Molecule; Small Interfering RNA; Source; Stem cells; Stretching; Surface; System; Testing; Thick; Tissues; Transforming Growth Factor beta; Tumor Angiogenesis; Tumor-Derived; Variant; Vascularization; Viral; Work; angiogenesis; crosslink; fibrillogenesis; implantation; improved; in vivo; inhibitor/antagonist; malignant breast neoplasm; neoplastic cell; novel; novel strategies; physical science; research study; response; tool; tumor; tumor growth; tumorigenesis

DESCRIPTION (provided by applicant): Increased tissue stiffness represents a hallmark of breast cancer that is mediated by physicochemical alterations of the extracellular matrix (ECM); however, the mechanisms through which enhanced ECM stiffness promotes tumor angiogenesis, and hence growth, are poorly understood. This project investigates the hypothesis that paracrine signaling by breast cancer cells increases fibronectin (Fn) matrix assembly by adipose-derived stem cells (ASCs), thereby enhancing the pro-angiogenic capability of both ASCs and endothelial cells to promote tumor vascularization. To investigate this hypothesis we propose a combination of biochemical and physical science approaches that will enable us to quantify the impact of tumor-derived soluble factor signaling on the conformation and rigidity of ASC-deposited Fn matrices. Specifically, we will use Fluorescence Resonance Energy Transfer (FRET) imaging and the Surface Forces Apparatus (SFA) to measure Fn mechanics at the macromolecular and cell/tissue level, respectively, and will assess the impact of these parameters on pro-angiogenic signaling in vitro and in vivo. This work will be accomplished in three specific aims: In Aim 1, we will evaluate Fn matrix assembly by ASCs in the presence or absence of tumor cell- conditioned media and identify signaling molecules contributing to these changes. In Aim 2, we will analyze the contributions of ASC-regulated Fn matrix characteristics towards a tumor-associated, pro-angiogenic phenotype of ASCs and endothelial cells. In Aim 3, we will determine whether ASC-regulated Fn matrix assembly promotes tumor angiogenesis, stiffness, and growth in vivo and evaluate the contributions of the signaling molecules identified in aim 1 in this pathogenesis. Transforming growth factor beta (TGF-beta) signaling will be the initial focus of the proposed studies, as this factor modulates tumorigenesis, cell contractility, and Fn assembly. Additionally, we anticipate identification of novel factors already implicated in Fn mechanics yet with an undefined role in tumor vascularization. By correlating Fn conformation and mechanics with pro-angiogenic signaling in the tumor microenvironment this work will broadly impact our understanding of the connection between tumor stiffness and vascularization and may lead to the identification of novel anti- angiogenic targets and improved therapies. While the emphasis in the proposed studies is to determine the role of ASCs in this process, a variety of other physiological and pathological situations critically rely upon ECM mechanics (e.g., organogenesis, atherosclerosis). The culture systems and mechanical testing strategies developed as part of this project introduce radically new approaches to investigate these processes. PUBLIC HEALTH RELEVANCE: Sustained angiogenesis is a hallmark of breast cancer that is influenced by extracellular matrix (ECM) mechanics; however, it remains unclear whether or not fibronectin (Fn) matrix assembly by tumor-associated adipose-derived stem cells (ASCs) may play a role in this process. This research will integrate biochemical and physical science tools to determine the effect of tumor-derived soluble factors on the rigidity of ASC-deposited Fn matrices and evaluate if these changes promote tumor vascularization. This interdisciplinary strategy has the potential to not only revolutionize our understanding of tumor angiogenesis, but also to provide widely applicable approaches to study other physiological and pathological processes that depend on Fn mechanics.

描述（由申请人提供）：增加的组织刚度代表了乳腺癌的标志，该标志是通过细胞外基质（ECM）的理化改变介导的；然而，对ECM刚度增强的机制促进了肿瘤血管生成及其生长的理解很少。该项目研究了以下假设：乳腺癌细胞的旁分泌信号传导通过脂肪衍生的干细胞（ASC）增加了纤连蛋白（FN）基质组装，从而增强了ASCS和内皮细胞的前血管生成能力，以促进肿瘤血管化。为了研究这一假设，我们提出了生化和物理科学方法的结合，这将使我们能够量化肿瘤衍生的可溶性因子信号传导对ASCESPOSEDISED FN矩阵构象和刚性的影响。具体而言，我们将分别使用荧光共振能量转移（FRET）成像和表面力设备（SFA）在大分子和细胞/组织水平上分别测量FN力学，并评估这些参数对体外和体内体内促血管生成信号的影响。这项工作将以三个特定的目的来完成：在AIM 1中，我们将在存在或不存在肿瘤细胞培养基的情况下通过ASC评估FN矩阵组装，并确定有助于这些变化的信号分子。在AIM 2中，我们将分析ASC调节的FN矩阵特征对ASC和内皮细胞的肿瘤相关，亲血管生成的表型的贡献。在AIM 3中，我们将确定ASC调节的FN矩阵组装是否促进体内肿瘤血管生成，僵硬和生长，并评估AIM 1中在本发病机理中鉴定出的信号分子的贡献。转化生长因子β（TGF-β）信号传导将是拟议研究的初始焦点，因为该因子调节了肿瘤发生，细胞收缩力和FN组装。此外，我们预计鉴定已经与FN力学有关的新因素鉴定，但在肿瘤血管中具有不确定的作用。通过将FN构象和力学与肿瘤微环境中的促血管生成信号传导相关联，这项工作将广泛影响我们对肿瘤僵硬与血管形成之间联系的理解，并可能导致鉴定出新型的抗血管生成靶标和改善治疗方法。拟议的研究的重点是确定ASC在此过程中的作用，但许多其他生理和病理状况严重依赖ECM力学（例如，器官发生，动脉粥样硬化）。作为该项目的一部分而开发的培养系统和机械测试策略引入了从根本到研究这些过程的新方法。 公共卫生相关性：持续的血管生成是受细胞外基质（ECM）力学影响的乳腺癌的标志；但是，目前尚不清楚通过肿瘤相关的脂肪衍生的干细胞（ASC）是否在此过程中起作用。这项研究将整合生化和物理科学工具，以确定肿瘤衍生的可溶性因子对ASC沉积FN矩阵刚度的影响，并评估这些变化是否促进了肿瘤血管形成。这种跨学科策略不仅有可能彻底改变我们对肿瘤血管生成的理解，而且还提供了广泛适用的方法来研究依赖FN力学的其他生理和病理过程。