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.

项目概要/摘要 人体器官系统需要在宏观尺度上在时间和空间上协调的多细胞行动 启动、维持或终止专用且重要的功能。包含离散或分布式的单元 无法协调地对适当刺激做出反应的生理系统可能会导致严重的发病率 并常常导致死亡。集体和协调的生理活动通常涉及数百万至数十亿个细胞 可能跨越很长的物理距离。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其潜在机制至关重要 疾病并开发治疗方法。然而，目前还没有技术可以快速量化 细胞集合内所有细胞对传输的远端扰动的机械细胞反应。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）用于并行和直接测量超过 1.0 x 的细胞牵引力 具有细胞空间分辨率的 1.0 cm2 视野 (FOV) 和 1,000 帧/秒的时间分辨率。点 提供比传统牵引力显微镜大 4 个数量级的 FOV。心肌细胞 （CM）是这里的试验台，因为它对心血管疾病具有很高的影响潜力，而心血管疾病是主要原因 西方世界的死亡率。我们将展示 SPOT 确定定量指标的能力 健康和患病状态下人类 CM 收缩和舒张的异常。我们将建立证据 SPOT 筛选增强或影响桥粒斑蛋白 CM 收缩的小分子的概念研究 不足的国家。我们将建立一个集成 SPOT 的平台，用于直接收缩力测量和 用于 CM 片材电性能测量的光学测绘。这将首次使 单细胞CM片时空机电耦合行为研究 解决。我们将区分 CM 的不同亚型、它们的分布、它们的相互作用以及它们的作用。 外部扰动下的表型反应。我们将应用这个平台来研究结构 通过整合定量生物量和 hESC 衍生的 CM 的机电耦合特性 使用非侵入性活细胞干涉测量法 (LCI) 测量的刚度数据。生物量和细胞硬度的变化 是可药物化的生物物理参数，与 CM 的机械收缩/松弛周期相关。在 除了对可能影响美国公民头号杀手的CM进行详细研究外，SPOT 应用程序应该具有实用性，并在需要细胞或组织的其他环境中提供新的见解 牵引力的产生。此类设置可能包括培养皿中的伤口愈合模型、癌细胞模型 转移或影响细胞和组织结构完整性（例如结缔组织）的疾病模型 埃勒斯-当洛斯或马凡综合征。