High throughput platform for simultaneous multiparametric assessment of cardiac physiology for heart failure drug development

用于心力衰竭药物开发的心脏生理学同步多参数评估的高通量平台

基本信息

批准号：
10745000
负责人：
MARK MERCOLA
金额：
$ 46.32万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10745000
关键词：
Acceleration Adherent Culture Adoption Adult Affect Assessment tool Automation Basic Science Benchmarking Biological Biological Assay Biological Products Calcium Cardiac Cardiac Myocytes Cardiomyopathies Cell Line Cell model Cells Clinical Code Colony-Forming Units Assay Computer software Data Data Analyses Data Set Dependence Development Diastolic heart failure Disease Disease model Drug Screening Fluorescence Generations Genes Genetic Genetic study Heart Diseases Heart failure Heterogeneity Human Image Individual Kinetics Measurement Measures Methods Microscopy Modeling Modernization Myocardial dysfunction Patients Performance Pharmaceutical Preparations Physiology Population Preparation Procedures Process Proteins Protocols documentation Readiness Reagent Relaxation Resolution Rest Run-On Assays Scientist Standardization Systolic heart failure Technology Technology Assessment Time Traction Force Microscopy Validation Viral Viral Vector Visualization biopharmaceutical industry calcium indicator cell type computer program computerized tools disease phenotype drug candidate drug development drug discovery drug use screening feature detection genetic variant graphical user interface heart function imaging system individual patient induced pluripotent stem cell insight instrumentation mechanical force meter millisecond mortality novel therapeutics ratiometric sensor skills stable cell line stem cell model tool user-friendly

项目摘要

Heart failure affects 2-3% of the US population and remains the single largest cause of mortality. Despite the large unmet need, heart failure drug development is notoriously difficult, and few first-in- class drugs have been approved in the past decade. Human induced pluripotent stem cell (hiPSC)- based models of heart disease are widely considered to hold tremendous potential for the development of heart failure drugs because they faithfully model disease phenotypes and reflect individual patient genetics. However, their utility for drug screening is limited because the technology for assessing disease modifying effects are too cumbersome and low throughput for large-scale screens. Visualizing disease-modifying activity of genes and drugs for heart failure requires kinetic read outs of cardiomyocyte function that correlate calcium (Ca2+) cycling with contractile force and resting tension to reveal systolic and diastolic heart dysfunction. The lack of off-the-shelf solutions for simultaneous measurement of these parameters is a critical gap that has hindered the pace of basic research into disease mechanisms and drug development. Although modern high content imaging systems can acquire the requisite fast kinetic datasets, the major roadblock is that available data analysis tools lack key capabilities, are too low throughput, and/or require substantial coding expertise to implement in large-scale genetic studies and drug discovery. Resolving this roadblock will be transformative by placing powerful tools in the hands of scientists without coding expertise, enabling them to develop gene and drug screens using iPSC and adult cardiomyocyte models of heart failure. We will deliver an integrated toolbox of software, reagents, and standardized protocols for contemporaneous measurement of intracellular and subcellular Ca2+ dynamics with contractile force and resting tension that can be overlaid with subcellular feature detection – all compatible with 384-well plate format – to model systolic and diastolic heart function. The software will have a user-friendly graphical user interface to fully automate measurements of Ca2+- contractility force curves (in absolute µM and nN terms) from beating cardiomyocytes. A key feature will be individual, cell-by-cell analysis that will increase dynamic range and allow the recognition of cellular heterogeneity in the preparations making possible realistic culture models with multiple cell types. We will also develop a toolkit of viral vectors to deliver genetically encoded sensors of absolute intracellular Ca2+ concentrations in cardiomyocyte populations without generating stable cell lines. The integrated platform will be validated and benchmarked against current software in pilot drug and gene screens that demonstrate ability to quantify disease-modifying activities for systolic and diastolic heart disease. Quantitative performance measures will evaluate assay readiness.

心力衰竭影响着 2-3% 的美国人口，并且仍然是导致死亡的最大原因。尽管存在大量未满足的需求，但心力衰竭药物的开发是出了名的困难，而且很少有首创药物。人类诱导多能干细胞（hiPSC）在过去十年中已获得批准。基于心脏病的模型被广泛认为具有巨大的发展潜力心力衰竭药物，因为它们忠实地模拟疾病表型并反映个体患者然而，由于评估技术有限，它们在药物筛选中的效用有限。对于大规模屏幕来说，疾病修饰效果过于繁琐且通量低。可视化基因和药物治疗心力衰竭的疾病修饰活性需要动力学读数将钙 (Ca2+) 循环与收缩力和静息张力相关的心肌细胞功能揭示收缩性和舒张性心脏功能障碍缺乏现成的解决方案。这些参数的测量是一个关键差距，阻碍了基础研究的步伐尽管现代高内涵成像系统可以帮助疾病机制和药物开发。获取必要的快速动力学数据集，主要障碍是缺乏可用的数据分析工具关键功能，吞吐量太低，和/或需要大量的编码专业知识才能在大规模遗传学研究和药物发现。通过向科学家提供强大的工具来解决这一障碍将是变革性的无需编码专业知识，使他们能够使用 iPSC 和成人开发基因和药物筛选我们将提供一个包含软件、试剂和工具的集成工具箱。同时测量细胞内和亚细胞 Ca2+ 的标准化方案具有收缩力和静息张力的动力学，可以与亚细胞特征叠加检测 - 全部与 384 孔板格式兼容 - 用于模拟心脏收缩和舒张功能。该软件将具有用户友好的图形用户界面，可全自动测量 Ca2+- 跳动心肌细胞的收缩力曲线（以绝对 µM 和 nN 表示）。将进行单独的逐个细胞分析，这将增加动态范围并允许识别制剂中的细胞异质性使得具有多个细胞的真实培养模型成为可能我们还将开发一个病毒载体工具包，以提供绝对的基因编码传感器。心肌细胞群中的细胞内 Ca2+ 浓度，而不产生稳定的细胞系。集成平台将根据试点药物和基因领域的当前软件进行验证和基准测试能够量化心脏收缩和舒张疾病缓解活动的屏幕定量性能测量将评估检测准备情况。