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DISEASE MODELING AND PHENOTYPIC DRUG SCREENING FOR DYSTROPHIC CARDIOMYOPATHY

营养不良性心肌病的疾病建模和表型药物筛选

基本信息

批准号：
10116566
负责人：
Deok-Ho Kim
金额：
$ 52.09万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-15 至 2023-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10116566
关键词：
3-Dimensional Achievement Address Adult Animal Testing Animals Architecture Biological Assay Biomedical Engineering CRISPR/Cas technology Cardiac Cardiac Myocytes Cardiomyopathies Cell Culture Techniques Cell Density Cell Survival Cells Characteristics Clinical Trials Complex Cues Cytoskeleton Data Development Disease Progression Disease model Drug Screening Drug Targeting Duchenne muscular dystrophy Dystrophin EFRAC Electrophysiology (science)Engineering Ensure Environment Exhibits Exposure to Extracellular Matrix Failure Functional disorder Generations Genes Goals Heart Heart Rate Human In Vitro Individual Investigation Kinetics L-Type Calcium Channels Left ventricular structure Measures Mediating Membrane Metabolic Methods MicroRNAs Microelectrodes Modeling Monitor Movement Muscular Dystrophies Myocardial Myocardial dysfunction Myocardium Nature Nitric Oxide Nuclear Receptors Organoids Oxidative Stress Pathway Pathology Pathway interactions Patients Pharmaceutical Preparations Phenotype Physiological Pluripotent Stem Cells Process Property Pump Receptor Signaling Relaxation Research Role Sampling Series Signal Pathway Signal Transduction Stimulus Stroke Volume Structure Symptoms System Techniques Technology Testing Thyroid Hormones Time Tissue Engineering Tissues Urine Validation Ventricular Work base combinatorial disease phenotype drug discovery drug efficacy dystrophic cardiomyopathy fetal heart function heart rate variability hormonal signals human model human stem cells human subject human tissue immunocytochemistry improved induced pluripotent stem cell microphysiology system nanopattern next generation novel organizational structure overexpression pre-clinical pressure response screening stem cells success therapy design three-dimensional modeling

项目摘要

PROJECT SUMMARY Goal: We propose to utilize a series of novel human iPSC-based cardiac microphysiological systems of increasing complexity to investigate the hypothesis that maturation of dystrophic cardiomyocytes is necessary to elucidate correct disease phenotype development in vitro. The aim is to create a multifaceted screening system using several core technologies developed by our group to evaluate different aspects of myocardial electromechanical function. We have developed the ability to engineer pluripotent stem cells from patient urine, enabling non-invasive cell sampling from human subjects. Furthermore, we have collected preliminary data demonstrating that application of combinatorial maturation stimuli to healthy and dystrophin-null cardiomyocytes helps stratify the disease phenotype. Based on these achievements, we posit that the use of a targeted set of functional assays in combination with appropriate maturation stimuli will provide a more comprehensive understanding of disease progression in muscular dystrophy. Focus/Aim: Our proposed research focuses on the use of techniques with the potential to act synergistically to enhance cardiac phenotype development in stem cell-derived cardiomyocytes through manipulation of different cellular mechanisms. Specifically, we will investigate the effect of structural organization by nanopatterned substrates, nuclear receptor signaling by thyroid hormone, and alterations in metabolic signaling pathways by Let-7 microRNA over-expression on the development of healthy cardiomyocytes and their dystrophin-null counterparts created using CRISPR-Cas9 gene editing technology. Nanotopographic microelectrode arrays will be used to evaluate electrophysiological function (Aim 1), while nanopatterned cell sheet stacking technology will be used to create 3D cardiac patches for analyzing contractile function (Aim 2) as well as organized 3D ventricle structures for assessing pressure generation and stroke volume (Aim 3). Each of these systems will be used to evaluate a panel of drugs for their potential to ameliorate the dystrophic phenotype. The compounds chosen for this study target a range of metabolic, structural, and signaling pathways known to be associated with different aspects of muscular dystrophy pathology. The analysis of multiple functional endpoints for each compound will therefore provide more comprehensive information on the likely effect of drugs when administered to human patients. The movement from platforms with higher levels of throughput to those with higher degrees of biomimicry, as the work transitions from Aim 1 to Aim 3, constitutes a natural “funneling” of the drug screening process. Candidates identified using simpler multiplexed models will be re- evaluated using systems that offer closer representations of the native tissue, and provide physiological endpoints analogous to those monitored in patients. As such, the proposed method for studying maturation and dystrophic phenotype development could provide the framework for a next generation screening process geared towards replacing animal testing with increasingly physiologically representative models of the myocardium.

项目总结