SPOTs: Optical Technologies for Instantly Quantifying Multicellular Response Profiles

SPOT：用于即时量化多细胞响应曲线的光学技术

• 批准号: 10160919
• 负责人: Pei-Yu Chiou
• 金额: $ 37.92万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10160919
• 关键词: Address; Affect; Age; Area; Arrhythmogenic Right Ventricular Dysplasia; Beds; Behavior; Biomass; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Cells; Chemicals; Collection; Color; Connective Tissue Diseases; Coupling; Data; Defect; Development; Disease; Disease model; Distal; Drug Screening; Electrophysiology (science); Fibroblasts; Future; Generations; Genes; Genetic Diseases; Giant Cells; Goals; Heart Atrium; Human; Imaging technology; Interferometry; Left; Machine Learning; Marfan Syndrome; Measurement; Measures; Mechanics; Methods; Modeling; Monitor; Morbidity - disease rate; Neoplasm Metastasis; Optics; Pathologic; Pharmacology Study; Phenotype; Physiological; Population; Process; Property; Regenerative research; Relaxation; Reporting; Resolution; Series; Spottings; Stimulus; Structure; System; Technology; Testing; Therapeutic; Time; Tissues; Traction; Traction Force Microscopy; Transplantation; Ventricular; Western World; biological systems; biophysical properties; body system; cancer cell; design; desmoplakin; electrical measurement; electrical property; human embryonic stem cell; human pluripotent stem cell; imaging platform; improved; indexing; insight; instrument; mechanical properties; mortality; new technology; patch clamp; prospective; public health relevance; response; screening; small molecule; technology development; temporal measurement; tool; wound healing

PROJECT SUMMARY/ABSTRACT Human organ systems require temporally and spatially coordinated multicellular actions at a macroscale to actuate, sustain, or terminate dedicated and vital functions. Cells that comprise discrete or distributed physiologic systems that fail to respond to appropriate stimuli with coordination may cause significant morbidity and often mortality. Collective and coordinated physiologic activities typically involve millions to billions of cells that may span large physical distances. Technologies for quantifying the electrical, chemical, and mechanical coupling in these multicellular systems are critically important to understanding the underlying mechanisms of disease and develop therapeutic approaches. However, no technology currently exists to quantify rapid mechanical cell responses to transmitted distal perturbations for all cells within a collection of cells. This multi- PI proposal (Chiou (contact PI) and Teitell) aims to develop a new platform imaging technology called SPOT (single pixel optical technology) for concurrent and direct measurements of cellular traction forces over a 1.0 x 1.0 cm2 field of view (FOV) with cellular spatial resolution, and a 1,000 frames/sec temporal resolution. SPOT provides a 4-order of magnitude larger FOV than conventional traction force microscopy. Cardiomyocytes (CMs) are the test bed here because of a high potential for impact in cardiovascular disease, the leading cause of mortality in the Western World. We will demonstrate the ability for SPOT to determine quantitative indices of abnormalities for human CM contraction and relaxation in healthy and diseased states. We will establish proof of concept studies in SPOT screens for small molecules that augment or affect CM contraction in desmoplakin deficient states. We will build a platform that integrates SPOT for direct contraction force measurements and Optical Mapping for electrical property measurements for sheets of CMs. This will enable, for the first time, studies of temporal and spatial electromechanical coupling behaviors for sheets of CMs at single cell resolution. We will distinguish different subtypes of CMs, their distributions, their interactions, and their phenotypic responses under external perturbations. And we will apply this platform to investigate the structural and electromechanical coupling properties of hESC-derived CMs by integrating quantitative biomass and stiffness data measured using non-invasive live cell interferometry (LCI). Changes in biomass and cell stiffness are druggable biophysical parameters with correlates to mechanical contraction/relaxation cycles of CMs. In addition to detailed studies of CMs that have the potential to impact the number one killer of US citizens, SPOT applications should have utility and provide new insights in additional settings that require cell or tissue traction-force generation. Such settings could include models in a dish for wound healing, cancer cell metastasis, or models of diseases that affect cell and tissue structural integrity, such as connective tissue disorders Ehlers-Danlos or Marfan syndromes.

项目摘要/摘要 人体器官系统需要在宏观上进行时间和空间协调的多细胞作用 促使，维持或终止专门和重要的功能。包含离散或分布的单元 无法通过协调响应适当刺激的生理系统可能会导致明显的发病率 而且经常死亡。集体和协调的生理活动通常涉及数百万个细胞 这可能跨越大的物理距离。量化电气，化学和机械的技术 这些多细胞系统中的耦合对于理解的基本机制至关重要 疾病并发展治疗方法。但是，目前尚无技术来量化快速 机械细胞对细胞集合中所有细胞的远端扰动的机械响应。这个多 PI提案（Chiou（联系PI）和Teitell）旨在开发一种名为Spot的新平台成像技术 （单像素光学技术）用于在1.0 x上同时和直接测量细胞牵引力 1.0 CM2视野（FOV）具有细胞空间分辨率，并具有1,000帧/秒的时间分辨率。点 与传统牵引力显微镜相比，提供了4阶的大小FOV。心肌细胞 （CMS）是这里的测试床，因为对心血管疾病的影响很高，这是主要原因 西方世界的死亡率。我们将展示点确定定量指数的能力 健康和患病状态的人类CM收缩和放松的异常。我们将建立证据 小分子的概念研究，这些分子增加或影响脱莫普拉金的CM收缩 状态不足。我们将建立一个平台，该平台集成了直接收缩力量测量的位置和 用于CMS板的电气测量的光学映射。这将首次实现 对单细胞处CMS板的时间和空间机电耦合行为的研究 解决。我们将区分CMS的不同子类型，它们的分布，它们的互动及其 外部扰动下的表型反应。我们将应用此平台调查结构 通过整合定量生物质和 使用非侵入性活细胞干涉法（LCI）测量的刚度数据。生物量和细胞刚度的变化 是可药物的生物物理参数，与CMS的机械收缩/松弛周期相关。在 除了对CM的详细研究外，有可能影响美国公民的第一杀手 应用应具有实用性，并在需要细胞或组织的其他设置中提供新的见解 牵引力产生。这样的设置可能包括在菜肴中包括伤口愈合，癌细胞的模型 转移或影响细胞和组织结构完整性的疾病模型，例如结缔组织 疾病ehlers-danlos或Marfan综合征。