An in vitro electrophysiology system for high-throughput measurement of cardiomyocyte action potential

用于高通量测量心肌细胞动作电位的体外电生理系统

• 批准号: 10759677
• 负责人: XIN JIANG
• 金额: $ 34.32万
• 依托单位: CYION TECHNOLOGIES, LLC
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10759677
• 关键词: 3-Dimensional; Accounting; Action Potentials; Algorithmic Software; Biological Assay; Biological Markers; Cardiac; Cardiac Electrophysiologic Techniques; Cardiac Myocytes; Cardiovascular system; Cause of Death; Cessation of life; Collaborations; Communication; Computer software; Data Analyses; Development; Electrodes; Electrophysiology (science); Foundations; Generations; Heart Diseases; Heterogeneity; Human; In Vitro; Individual; Industry; Investigation; Measurement; Measures; Methods; Myocardial dysfunction; Nanotechnology; Nature; Optical Methods; Optics; Phase; Process; Protocols documentation; Publishing; Reader; Records; Research Personnel; Risk; Role; Safety; Sampling; Signal Transduction; Support System; System; Techniques; Technology; Time; United States; Universities; Validation; Work; commercialization; cost; data acquisition; data quality; design; drug development; drug testing; extracellular; fabrication; high standard; high throughput screening; large datasets; lithography; nano; nanoelectrode array; nanoelectrodes; novel; patch clamp; programs; public health relevance; response; solid state; success

Summary Heart disease has been the #1 cause of fatality in the United States since 1950. Additionally, cardiovascular safety liability especially pro-arrhythmic risk remains a leading cause for the high attrition rate in drug development. For investigation of mechanisms and potential therapies for cardiac dysfunction, there has been strong demand from industry and academic alike for high quality, high-throughput assay of cardiac action potential (AP), a powerful biomarker for cardiac dysfunction. Unfortunately, existing techniques for measurement of AP, including electrical and optical methods, all suffer from severe limitations and are technically challenging and/or costly to execute at scale. These limitations are further compounded by the profound heterogeneity of human-derived cardiomyocytes. This Phase I proposal aims to develop a novel in vitro electrophysiology system that supports high-throughput measurement of intracellular AP (iAP) using human-derived cardiomyocytes. More specifically, we seek to demonstrate the feasibility for researcher even with limited expertise in electrophysiology to perform high-throughput iAP assay that includes data acquisition and analysis. Central to this electrophysiology system is a 3D nanoelectrode array (NEA) developed at Stanford University, which bridges the gap between intracellular and extracellular electrophysiology and enables high-quality and scalable recordings of iAP from many individual cardiomyocytes. Based on this NEA technology, Cyion Technologies has previously developed a first-generation electrophysiology system (Vincent) that records iAP signals from up to 60 discrete cardiomyocytes in a single-well NEA substrate. Built on this success, in this proposal we seek to develop an additional “plate-reader” option that measures 672 individual cardiomyocytes using a 96-well NEA substrate. We will also implement a software algorithm in user software for automated, batch analysis of iAP signals, which is necessary for higher-throughput assay.

总结 自1950年以来，心脏病一直是美国死亡的头号原因。心血管安全性 责任，特别是药物风险仍然是药物开发中高流失率的主要原因。为 研究心功能不全的机制和潜在的治疗方法， 心脏动作电位（AP）的高质量、高通量测定， 心脏功能障碍的生物标志物。不幸的是，现有的用于测量AP的技术，包括电测量和 光学方法都受到严重的限制，并且在技术上具有挑战性和/或大规模执行成本高。 这些局限性由于人源性心肌细胞的高度异质性而进一步加剧。 该第一阶段提案旨在开发一种新的体外电生理学系统， 使用人源性心肌细胞测量细胞内AP（iAP）。更具体地说，我们力求 证明了研究人员即使在电生理学方面的专业知识有限，也可以进行高通量研究的可行性 iAP测定，包括数据采集和分析。 该电生理系统的核心是斯坦福大学开发的3D纳米电极阵列（NEA）， 弥合细胞内和细胞外电生理学之间的差距， 来自许多个体心肌细胞的iAP记录。基于这种NEA技术，Cyion Technologies 之前开发了第一代电生理系统（Vincent），可记录高达60 单个孔NEA基底中的离散心肌细胞。在这一成功的基础上，我们在本提案中寻求制定一项 额外的“平板读取器”选项，使用96孔NEA基板测量672个单独的心肌细胞。我们将 我还在用户软件中实现软件算法，以便对iAP信号进行必要的自动批量分析 用于更高通量的测定。