Multiplexed quantitative measurements of field potential and contractility on biomimetically-matured hPSC-CMs

对仿生成熟 hPSC-CM 的场电位和收缩性进行多重定量测量

• 批准号: 9910089
• 负责人: Nicholas Andrew Geisse
• 金额: $ 22.5万
• 依托单位: CURI BIO INC
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-16 至 2020-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9910089
• 关键词: Address; Adoption; Arrhythmia; Biochemical; Biological; Biological Assay; Biomimetics; Cardiac; Cardiac Myocytes; Cardiotoxicity; Cell Culture Techniques; Cell Shape; Cells; Clinical Trials; Communities; Consumption; Cost efficiency; Cues; Data; Detection; Development; Devices; Drug Screening; Electrical Resistance; Electrodes; Electrophysiology (science); End Point Assay; Environment; Extracellular Matrix; Failure; Frequencies; Geometry; Health; Heart; Human; In Vitro; Industry Standard; Measurable; Measurement; Measures; Mechanics; Methods; Microelectrodes; Microscopy; Mission; Morphology; Optics; Pattern; Pharmaceutical Preparations; Phase; Phenotype; Physiological; Platinum; Polymers; Process; Property; Proxy; Research; Resources; Speed; Stem cells; Stretching; Structure; Surface; Techniques; Technology; Testing; Therapeutic; Thinness; Time; Tissues; Traction Force Microscopy; Translations; Work; base; bench to bedside; biomaterial compatibility; cost; design; drug candidate; drug development; electric impedance; electrical measurement; extracellular; high throughput screening; improved; induced pluripotent stem cell; innovation; instrument; mechanical properties; medication safety; nanofabrication; nanofiber; nanopattern; novel; novel therapeutics; polydimethylsiloxane; pre-clinical; response; scale up; sensor; success; tissue culture; tool

PROJECT SUMMARY The development of new drugs is a costly and time-consuming process (~$2.5 billion over 5-10 years) with a very low success rate where only 1 out of 10,000 candidates will ever reaches the market. One of the leading causes for this issue is the cardiotoxicity of drug candidates, wherein a drug has an off-target effect of causing cardiac arrhythmias. As a result, significant effort and resources have been allocated to create more predictive preclinical and in vitro drug screening platforms. Human derived induced pluripotent cardiac myocyte stem cells (hPSC-CMs) are a promising tool to address this problem, but their relative lack of phenotypic maturity remains a barrier to their wide adoption. Some platforms focus on mimicking the structural (e.g. biomimetic cultureware), mechanical (e.g. cell and tissue stretching devices), and electrochemical (e.g. microelectrode array platforms) cues of the extracellular matrix of the tissue to improve hPSC-CM maturity. While these cues are vital to the tissue development, they are oftentimes incompatible with the high-throughput assays that are required by drug developers. Further, measuring maturity within an assay is a challenge. Contractility is considered to be a highly- accurate method of measuring maturity, state of differentiation, and general health of cardiomyocytes. The currently available measurement tools cardiomyocytes (CMs) contractility can be generally grouped as either impedance-based or microscopy-based (such as traction force microscopy; TFM). Impedance-based measurements are often fast and accurate but lacking in terms of capturing quantitative information, as impedance measures only cell shape changes and uses that as proxy of the cell contraction. In contrast, TFM techniques are capable of quantifying CM contraction, but it is laborious and incompatible with high-throughput platforms. Indeed, a critical need of the research community is a multiplexed platform that measures contractility in a high-throughput and quantitative fashion in an environment that applies extracellular cues to drive the development and maturity of CMs. NanoSurface Biomedical’s mission is to develop a first-of-its kind microelectrode array device that provides a biomimetic culture environment and is multiplexed with quantitative contractility measurement. We term this device the “MP-ForceMEA”. The MP-ForceMEA will use an innovative strain-gauge sensor with an MEA platform and will represent a novel instrument capable of simultaneous detection of electrophysiology and contractility in a highly parallel, high-throughput, and scalable manner. Phase 1 activities will result in the development of a single-well novel platform compatible with standard end-point assays, and this work will then serve as the basis for progression into Phase 2, where the device will be scaled up to high-throughput assay formats. The resulting work will greatly improve the cost, efficiency, and safety of drug development and speed to market new lifesaving drugs.

项目摘要 新药的开发是一个昂贵而耗时的过程（5-10年约25亿美元）， 成功率非常低，每10，000名候选人中只有1人能够进入市场。领先的 这个问题的原因是候选药物的心脏毒性，其中药物具有引起心脏病的脱靶效应。 心律失常因此，已经分配了大量的工作和资源来创建更有预测性的 临床前和体外药物筛选平台。人源性诱导多能心肌干细胞 hPSC-CM是解决这一问题的有希望的工具，但它们相对缺乏表型成熟度， 这是它们被广泛采用的障碍。一些平台专注于模仿结构（例如仿生培养皿）， 机械（例如细胞和组织拉伸装置）和电化学（例如微电极阵列平台） 提示组织的细胞外基质以改善hPSC-CM成熟。虽然这些线索对 组织发育，它们通常与药物开发所需的高通量测定不相容。 开发者此外，在测定中测量成熟度是一个挑战。收缩力被认为是一个高度- 准确测量心肌细胞成熟度、分化状态和总体健康状况的方法。的 目前可用的测量工具心肌细胞（CM）收缩性通常可以分为 基于阻抗或基于显微镜（例如牵引力显微镜; TFM）。基于阻抗 测量通常是快速和准确的，但缺乏捕获定量信息， 阻抗仅测量细胞形状变化，并将其用作细胞收缩的代表。相比之下，TFM 技术能够量化CM收缩，但它是费力的并且与高通量不相容。 平台事实上，研究界的一个关键需求是一个测量收缩性的多路复用平台 以高通量和定量的方式，在应用细胞外因子来驱动细胞外信号的环境中， CM的发展和成熟。NanoSurface Biomedical的使命是开发一种 微电极阵列装置，其提供仿生培养环境，并且与定量微电极阵列装置多路复用。 收缩性测量。我们将该器械称为“MP-ForceMEA”。MP-ForceMEA将采用创新的 应变片传感器与MEA平台，并将代表一种新的仪器，能够同时 以高度并行、高通量和可扩展的方式检测电生理学和收缩性。相 1项活动将导致开发与标准终点兼容的单井新型平台 这项工作将作为进展到第二阶段的基础，在第二阶段，该设备将被扩展 直至高通量测定形式。由此产生的工作将大大提高成本，效率和安全性， 药物开发和速度，以市场新的救生药物。