MASS PRODUCTION OF PERSONALIZED HUMAN ENGINEERED HEART TISSUES

描述（由申请人提供）：心肌病有多种形式，包括肥厚型心肌病（HCM），1/500的患病率，可能导致运动员心脏性猝死;扩张型心肌病（DCM），1/5000的患病率，导致左心室扩大;致心律失常性右心室发育不良/心肌病（ARVD/C），1/1000的患病率，导致心脏纤维化。美国医疗保健系统每年用于心血管疾病治疗的费用超过2000亿美元.心血管治疗的疗效和毒性反应的个体间差异相当大。安全有效的心血管药物可以通过伴随诊断和新药的共同开发来实现。目的：开发体外疾病模型，在使用患者细胞产生的工程心脏组织（EHT）中重现个体患者的心肌病。细胞、组织培养平台和新型心肌细胞分化方案的自动化将降低成本，提高生产率和重现性，从而生成用于药物筛选和诊断开发的疾病模型。基本原理：人类iPSC技术将“盘中疾病”的想法变为现实，用于个性化医疗。然而，iPSC技术的成本减缓了它的进展。自动化干细胞培养与经济的机器人技术生产的药物筛选成本效益的患者特定的样品。近期目标：将威斯康星州大学Palecek博士的学术实验室开发的心脏分化方案转化为商业上可行的形式。将与威斯康星州医学院的Strande博士合作，评估使用从肌营养不良患者中分离的细胞制造EHT的系统的可行性。拟议项目：目的一是建立高通量的人心肌细胞分化优化系统，并实现EHT的自动化批量生产。目的2是证明基于EHT的肌营养不良模型的开发。衡量成功：将通过以下方式进行测量：1）从一批干细胞培养物中可再现地大量生产约400 EHT产品; 2）使用大量生产的CM生产生理学相关EHT; 3）患者心脏病表型的重建。附加影响：高通量干细胞培养条件筛选平台可应用于干细胞研究的所有项目。通过技术改造，可以获得足够的心肌细胞用于临床细胞或组织工程移植。估计十亿个细胞是治疗梗塞后损伤的足够数量的细胞。

DESCRIPTION (provided by applicant): Cardiomyopathy has various forms including hypertrophic cardiomyopathy (HCM), 1 in 500 prevalence, could cause sudden cardiac death in athletes, Dilated Cardiomyopathy (DCM), 1 in 5000 prevalence, results in an enlarge left ventricle, Arrhythmogenic Right Ventricular Dysplasia/Cardiomyopathy (ARVD/C), 1 in 1000 prevalence, causes cardiac fibrosis. Yearly over $ 200 billion is spent by American healthcare system for cardio- vascular treatment. Inter-individual variability in efficacy and toxicity respone is rather large for cardiovascular treatments. Safe and effective cardiovascular medicine can be achieved by co-development of companion diagnostics and novel drugs. Objective: To develop in vitro disease model that recapitulate individual patient's cardiomyopathy in engineered heart tissues (EHTs) generated using the patient's cells. The automation of cell, tissue culture platform, and novel cardiomyocytes differentiation protocol will reduce cost, increase productivity and reproducibility of generating those disease models for drug screening and diagnostics development. Rationale: The human iPSC technology moved "Diseases in a dish" idea to reality for personalized medicine. Yet the cost of iPSC technology slowed its progress. Automated stem cell culture with an economic robotics produces patient-specific samples for drug screening cost effectively. Immediate goal: To translate cardiac differentiation protocol developed in the academic labs of Dr. Palecek at University of Wisconsin into a commercially viable format. Feasibility of using the system for EHT fabrication with cells isolated from muscular dystrophy patients will be evaluated in collaboration with Dr. Strande at the Medical College of Wisconsin. Proposed project: Aim 1 is the development of high-throughput human cardiomyocytes differentiation optimization system and their automated mass-production for EHT fabrication. Aim 2 is to demonstrate EHT based muscular dystrophy model development. Measure of Success: It will be measured by 1) reproducible mass-production of ~400 EHT production from a batch of stem cell culture 2) production of physiologically relevant EHTs using mass produced CMs, 3) reconstitution of cardiac disease phenotype in patient. Additional Impact: High-throughput platform for screening stem cell culture conditions can be applied to all projects of stem cell research. A modification of technology could produce enough cardiac cells for cell or engineered tissue transplant in clinic. One billion cells were estimated to be a sufficient number of cells to treat post infarct injury.