Developing branch stress microscopy for the mechanobiology of 3D morphogenesis and invasive diseases

开发用于 3D 形态发生和侵袭性疾病的机械生物学的分支应力显微镜

• 批准号: 10710186
• 负责人: Cynthia A. Reinhart-King
• 金额: $ 19.06万
• 依托单位: UNIVERSITY OF ARKANSAS AT FAYETTEVILLE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-28 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10710186
• 关键词: 3-Dimensional; Animal Model; Biochemical; Biological; Biological Models; Biology; Biomedical Engineering; Cancer Model; Cell Communication; Cells; Communities; Complex; Computer Models; Computer software; Confocal Microscopy; Development; Developmental Biology; Disease; Elasticity; Engineering; Environment; Equation; Equilibrium; Evaluation; Extracellular Matrix; Fibroblast Growth Factor; Finite Element Analysis; Foundations; Future; Gland; In Vitro; Invaded; Kidney; Knowledge; Liquid substance; Lung; Malignant Neoplasms; Mammary gland; Maps; Measurement; Measures; Mechanical Stress; Mechanics; Methods; Microscopy; Modeling; Molecular; Morphogenesis; Morphology; Organ; Outcome; Output; Pathogenesis; Pathway interactions; Pattern; Performance; Play; Positioning Attribute; Procedures; Process; Proliferating; Property; Regenerative Medicine; Resources; Rewards; Risk; Risk Management; Role; Series; Shapes; Signal Pathway; Signal Transduction; Site; Solid; Somatotropin; Stress; Structure; Techniques; Technology; Tissues; Traction; Traction Force Microscopy; Width; Work; angiogenesis; design; developmental disease; experimental study; human disease; in silico; in vivo; insight; interest; mechanical force; mechanical signal; mechanotransduction; migration; novel; regenerative treatment; spatiotemporal; technology development; three-dimensional modeling; tool; transmission process; tumor; 3-Dimensional; Animal Model; Biochemical; Biological; Biological Models; Biology; Biomedical Engineering; Cancer Model; Cell Communication; Cells; Communities; Complex; Computer Models; Computer software; Confocal Microscopy; Development; Developmental Biology; Disease; Elasticity; Engineering; Environment; Equation; Equilibrium; Evaluation; Extracellular Matrix; Fibroblast Growth Factor; Finite Element Analysis; Foundations; Future; Gland; In Vitro; Invaded; Kidney; Knowledge; Liquid substance; Lung; Malignant Neoplasms; Mammary gland; Maps; Measurement; Measures; Mechanical Stress; Mechanics; Methods; Microscopy; Modeling; Molecular; Morphogenesis; Morphology; Organ; Outcome; Output; Pathogenesis; Pathway interactions; Pattern; Performance; Play; Positioning Attribute; Procedures; Process; Proliferating; Property; Regenerative Medicine; Resources; Rewards; Risk; Risk Management; Role; Series; Shapes; Signal Pathway; Signal Transduction; Site; Solid; Somatotropin; Stress; Structure; Techniques; Technology; Tissues; Traction; Traction Force Microscopy; Width; Work; angiogenesis; design; developmental disease; experimental study; human disease; in silico; in vivo; insight; interest; mechanical force; mechanical signal; mechanotransduction; migration; novel; regenerative treatment; spatiotemporal; technology development; three-dimensional modeling; tool; transmission process; tumor

PROJECT SUMMARY/ABSTRACT Branched structures are essential for the formation of many organs and glands during development. In addition, many invasive diseases including abnormal angiogenesis and collective cancer invasion also take the form of branches. Hence, understanding the mechanism underlying the patterning and morphogenesis of branches is of critical importance in both fundamental biology of development and treatment of human diseases. Branching processes, including the elongation, bifurcation, and termination of the branches, can be regulated by biochemical signals, such as fibroblast growth factors and hormones. Recent work also suggests that mechanical signals from the extracellular matrix and from neighboring cells also influence branching dynamics. However, likely due to the lack of quantitative tools that can measure the distribution of mechanical forces within the branches, how mechanics regulates the branching process is still not well understood. In this project, we propose to develop a novel quantitative tool, termed branch stress microscopy (BSM), that can precisely map the spatiotemporal distribution of intercellular mechanical stresses during the branching process. Even with significant developments in cell and tissue mechanics over the past decades, quantifying intercellular mechanical stresses within a three-dimensional space remains a challenging task. Hence, to manage the risk, the proposed project is designed with two progressively riskier and more rewarding aims. In Aim 1, we will develop a relatively simple 1D version of BSM that quantifies the cross-sectional stress along a morphogenetic branch. Confocal microscopy will be combined with a three-dimensional traction stress calculation to obtain the total force and average stress exerted at the cross section via force balance equations. We will then validate the stress calculated from 1D BSM against that from the current state of the art using 3D cancer collective migration as a biological model. In Aim 2, we will take one step further to develop a 3D version of BSM to resolve the complete 3D distribution of intercellular stresses within a branch segment. We will make necessary measurements and assumptions regarding the branch material properties and stress or displacement values at the boundary of the branch segment and turn the task into a boundary value problem in solid mechanics. We will then calculate the stress distribution within invading cancer branches using finite element analysis and validate the assumptions and the robustness of the tool by comparing with the stresses measured by the current state of the art. In sum, this project will combine in silico and in vitro engineering and biological approaches to develop a novel quantitative tool that may be widely applicable to any branching processes in vitro, ex vivo and even in vivo, thus providing a versatile technology for branching mechanobiology in development and diseases.

项目摘要/摘要 分支结构对于在发育过程中形成许多器官和腺体至关重要。此外， 许多侵入性疾病，包括异常血管生成和集体癌症侵袭，也采用 分支。因此，了解分支的模式和形态发生的基础机制是 在人类疾病的发展和治疗的基本生物学中，至关重要的重要性。分枝 包括分支机构的伸长，分叉和终止在内 生化信号，例如成纤维细胞生长因子和激素。最近的工作还表明机械 来自细胞外基质和相邻细胞的信号也会影响分支动力学。然而， 可能是由于缺乏可以测量机械力分布的定量工具所致 分支机构，如何调节分支过程的力学仍然不太了解。在这个项目中，我们建议 为了开发一种新型的定量工具，称为分支应力显微镜（BSM），可以精确映射 分支过程中细胞间机械应力的时空分布。即使有 在过去的几十年中，细胞和组织力学的重大发展，量化了细胞间力学 三维空间内的压力仍然是一项艰巨的任务。因此，为了管理风险，提议 项目的设计具有两个逐渐风险，更有意义的目标。在AIM 1中，我们将开发一个相对的 简单的1D版本的BSM量化沿形态发生分支的横截面应力。共焦 显微镜将与三维牵引力计算结合，以获得总力和 平均应力通过力平衡方程在横截面施加。然后我们将验证压力 使用3D癌集体迁移作为一个A作为A的1D BSM计算。 生物模型。在AIM 2中，我们将进一步开发一个3D版本的BSM来解决完整 分支段内细胞间应力的3D分布。我们将进行必要的测量和 关于分支材料特性以及应力或位移值的假设 分支段并将任务转变为固体力学中的边界价值问题。然后，我们将计算 使用有限元分析中的入侵癌症分支中的应力分布并验证假设 以及通过与当前最新状态测量的应力进行比较，使工具的鲁棒性。总而 该项目将在硅和体外工程和生物学方法中结合起来，以开发一种新颖的方法 定量工具可能广泛适用于体外，离体甚至体内的任何分支过程，因此 为开发和疾病中的分支机械生物学提供多功能技术。