MASS PRODUCTION OF PERSONALIZED HUMAN ENGINEERED HEART TISSUES

描述（由申请人提供）：心肌病有多种形式，包括肥厚性心肌病（HCM），发病率为1 / 500，可导致运动员心源性猝死；扩张性心肌病（DCM），发病率为1 / 5000，导致左心室增大；心律失常性右室发育不良/心肌病（ARVD/C），发病率为1 / 1000，导致心脏纤维化。美国医疗保健系统每年花费超过2000亿美元用于心血管治疗。在心血管治疗中，疗效和毒性反应的个体差异相当大。通过伴随诊断和新药的共同开发，可以实现安全有效的心血管医学。目的：建立一种体外疾病模型，在利用患者细胞生成的工程化心脏组织（EHTs）中再现个体心肌病。细胞、组织培养平台的自动化和新的心肌细胞分化方案将降低成本，提高生产效率和可重复性，以生成用于药物筛选和诊断开发的疾病模型。原理：人类iPSC技术将“疾病培养皿”的想法变成了个性化医疗的现实。然而，iPSC技术的成本阻碍了它的发展。自动化干细胞培养与经济机器人生产患者特异性样品的药物筛选成本有效。近期目标：将威斯康星大学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。我们将与威斯康星医学院的Strande博士合作，评估该系统用于肌肉萎缩症患者细胞EHT制造的可行性。拟建项目：目标1是开发用于EHT制造的高通量人心肌细胞分化优化系统及其自动化批量生产。目的2是展示基于EHT的肌营养不良模型的开发。成功的衡量：将通过以下方法来衡量：1)从一批干细胞培养中可重复批量生产约400 EHT； 2)使用批量生产的CMs生产生理相关的EHT； 3)重建患者的心脏病表型。附加影响：用于筛选干细胞培养条件的高通量平台可应用于所有干细胞研究项目。通过对技术的改进，可以产生足够的心脏细胞，用于临床细胞或工程组织移植。据估计，10亿个细胞足以治疗梗死后损伤。

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.