Optimization of electromechanical monitoring of engineered heart tissues

项目总结/摘要 心力衰竭和心律失常是遗传性心肌病的主要临床表现， 可能危及生命并导致这些疾病的显著发病率和死亡率。新患者/基因型- 针对不同形式的心肌病的特定疗法正在开发和临床试验中， 动物模型不能充分捕获临床相关的患者疾病表型和基因型。病人- 特定的人诱导多能干细胞（hiPSC）可以从容易获得的成体细胞（例如， 血液、皮肤或尿细胞）并分化成心脏样细胞、心肌细胞（hiPSC-CM）， 在人类心肌细胞背景下评估新治疗的机会。虽然hiPSC-CM模型 这些细胞是不成熟的，通常产生非生理性的， 产出工程化心脏组织（EHT）可以通过将hiPSC-CM浇铸到水凝胶基质中来产生， 由柔性柱支撑，以在培养皿中形成组织状结构。EHT提高细胞成熟度， 获取生理输出比如收缩力EHT中的心律失常建模，已经证明了更多 具有挑战性，只能通过侵入性的终端实验来完成，需要专门的染料和 成像设备。该项目旨在优化柔性电子技术在EHT中的应用 培养条件，使得收缩性和场电位可以作为电和机械活动来评估 同时且无创地用于不确定的时间段。该平台将使个性化药物 测试，如基因校正策略，目前正在开发中。我们开发了一个原型 采用这种技术的设备，我们称之为机电监测EHTs（emEHTs）。我们 本发明证明了emEHT用于捕获传感器阵列上的组织应变和场电位。 我们建议通过表征从制造到长期的设备功能来进一步验证这项技术。 文化，采用标准的产品开发质量改进过程，以确保一致的设备 质量和产出。为了更广泛地采用，将开发图形用户界面， 收集和分析设备输出。我们将通过先进的成像技术进一步验证这项技术 emEHT的钙和膜电压的表征。药理学研究评估了 将进行收缩性和促血管生成性试验，以证明药物试验和筛选的潜力。 先前收集和重编程的hiPSC来自携带已知高度表达的基因突变的患者。 将使用emEHTs评估致突变性，以证明建模遗传形式的潜力。 心肌病该项目将由心血管研究人员组成的跨学科团队共同领导， 具有临床心脏病学，心律失常建模，心血管遗传学技能的生物工程师，以及 微细加工该项目的成功完成将产生一个经过良好验证的工作emEHT平台 供具有hiPSC培养和分化方法经验的科学家使用。

PROJECT SUMMARY/ABSTRACT Heart failure and arrhythmias are the major clinical manifestations of genetic forms of cardiomyopathy, which can be life-threatening and cause significant morbidity and mortality in these diseases. New patient/genotype- specific therapies are in development and clinical testing for different forms of cardiomyopathy, however small animal models insufficiently capture the clinically relevant patient disease phenotypes and genotypes. Patient- specific human induced pluripotent stem cells (hiPSCs) can be created from readily available adult cells (e.g., blood, skin, or urine cells) and differentiated in heart-like cells, cardiomyocytes (hiPSC-CMs), offering opportunities to evaluate new treatments in a human cardiomyocyte context. While hiPSC-CM models are promising for disease modeling and drug testing, the cells are immature and often produce non-physiologic outputs. Engineered heart tissues (EHTs) can be created by casting hiPSC-CMs into hydrogel matrices, supported by flexible posts, to create tissue-like structures in the dish. EHTs improve cell maturity and provide access to physiologic outputs like force of contraction. Arrhythmia modeling in EHTs, has proven more challenging and has only been accomplished by invasive, terminal experiments requiring specialized dyes and imaging equipment. The project aims to optimize the application of flexible electronics technology to EHT culture conditions so that contractility and field potential can be evaluated as electrical and mechanical activity simultaneously and noninvasively for indefinite time periods. This platform will enable personalized drug testing like genetic correction strategies, which are currently in development. We developed a prototype device employting this technology, which we named electromechanically-monitored EHTs (emEHTs). We present demonstrating that emEHTs function to capture tissue strain and field potential on an array of sensors. We propose to further validate this technology by characterizing device function from fabrication to long-term culture, employing a standard product development quality improvement process to ensure consistent device quality and outputs. To allow for more generalized adoptability, a graphical user interface will be developed for collecting and analyzing device output. We will further validate this technology with advanced imaging characterization of calcium and membrane voltage of emEHTs. Pharmacologic studies that assess both contractility and arrhythmogenicity will be conducted to demonstrate potential for drug testing and screening. Previously collected and reprogrammed hiPSCs from patients harboring mutations in genes known to be highly arrhythmogenic will be assessed using emEHTs to demonstrate the potential of modeling genetic forms of cardiomyopathy. This project will be co-led by an interdisciplinary team of cardiovascular researchers and bioengineers with skill sets in clinical cardiology, arrhythmia modeling, cardiovascular genetics, and microfabrication. Successful completion of this project will result in a well-validated working emEHT platform for use by scientists with experience in hiPSC culture and differentiation methods.