Endothelial hiPSC-Cardiomyocyte Interactions Relevant to Model Disease and Aging

与模型疾病和衰老相关的内皮 hiPSC-心肌细胞相互作用

基本信息

批准号：
8851826
负责人：
MARK MERCOLA
金额：
$ 19.38万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-15 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8851826
关键词：
Action Potentials Adult Aftercare Aging Atomic Force Microscopy Biological Assay Biological Models Brightfield Microscopy Calcium Cardiac Myocytes Cell membrane Cells Charge Clinical Collaborations Computer software Conditioned Culture Media Data Detection Development Disease Disease model Endothelial Cells Evaluation Exclusion Exhibits Exploratory/Developmental Grant Filtration Fractionation Gene Expression Gene Proteins Generations Genes Goals Health Heparin Binding Homeostasis Individual Ion Channel Ion Pumps Lead Life Link Measures Mechanics Membrane Potentials Methods Metric Modeling Monitor Muscle Cells Optics Paracrine Communication Pharmaceutical Preparations Physiological Production Property Regulation Reporting Research Risk Assessment Signal Transduction Signaling Molecule Source Specific qualifier value Staging Stem cells Structure Technology Time Tissues Traction Translating Translational Research Vascular Endothelial Cell base cardiogenesis cellular imaging clinical application drug discovery drug sensitivity drug testing fetal high risk human embryonic stem cell imaging modality improved instrumentation liquid chromatography mass spectroscopy paracrine research study

项目摘要

DESCRIPTION (provided by applicant): In principle, differentiated cardiomyocytes derived from hESCs and hiPSCs are ideal model systems in which to study the physiological effects of disease and aging. However, hESC/hiPSC-derived cardiomyocytes characteristically exhibit electrical and mechanical properties of early fetal, rather than adult, cells. Although a few growt factor-like molecules are reported to have a maturational effect, little is known about the precise physiological effects or how the molecules can be used to improve the utility of hiPSC/hESC-derived cardiomyocytes to model aging and disease. The goal of this R21 proposal is to identify and characterize such molecules, based on preliminary data that endothelial cells normally produce maturation-inducing factors. Data are presented showing the development of instrumentation and software for high-throughput assessment of electrophysiological maturation. Using this technology, we provide preliminary evidence that endothelial cells (ECs) promote electrical and ion channel profile maturation of hiPSC/ESC-derived cardiomyocytes through the production of factors that accumulate in conditioned media. The specific aims of this proposal are to: 1) define EC-induced maturation by functional expression of ion channels and pumps, drug sensitivity profiling, action potential and calcium handling metrics, 2) determine if ECs also promote maturation of contractile apparatus structure and function, and 3) use the electrical and mechanical metrics from Aims 1 and 2 as quantitative metrics to identify and characterize the effects of EC-derived maturation factor(s). The identification of signals that promote cardiomyocyte maturation should lead to improved methods for deriving functional cardiomyocytes that will be enabling for studies of aging and disease. Beyond the high- risk/high-yield scope of the R21 project, we have established collaborations to evaluate whether the maturational factors indeed enhance the utility of hiPSC-cardiomyocytes as models for drug risk assessment and disease.

描述（由申请人提供）：原则上，源自 hESC 和 hiPSC 的分化心肌细胞是研究疾病和衰老的生理影响的理想模型系统。然而，hESC/hiPSC 衍生的心肌细胞典型地表现出早期胎儿细胞而非成人细胞的电学和机械特性。尽管据报道一些生长因子样分子具有成熟作用，但对其精确作用却知之甚少。生理效应或如何使用这些分子来提高 hiPSC/hESC 衍生的心肌细胞在模拟衰老和疾病方面的效用。 R21 提案的目标是根据内皮细胞通常产生成熟诱导因子的初步数据来识别和表征此类分子。数据显示了用于电生理成熟度高通量评估的仪器和软件的开发。利用这项技术，我们提供了初步证据，证明内皮细胞 (EC) 通过产生在条件培养基中积累的因子，促进 hiPSC/ESC 衍生的心肌细胞的电和离子通道谱成熟。该提案的具体目标是：1) 通过离子通道和泵的功能表达、药物敏感性分析、动作电位和钙处理指标来定义 EC 诱导的成熟，2) 确定 EC 是否也促进收缩装置结构和功能的成熟，和 3) 使用目标 1 和 2 中的电气和机械指标作为定量指标来识别和表征 EC 衍生成熟因子的影响。促进心肌细胞成熟的信号的识别应该会导致衍生功能性心肌细胞的改进方法，这将有助于衰老和疾病的研究。除了 R21 项目的高风险/高收益范围之外，我们还建立了合作，以评估成熟因素是否确实增强 hiPSC 心肌细胞作为药物风险评估和疾病模型的效用。