Optimization of electromechanical monitoring of engineered heart tissues

工程心脏组织机电监测的优化

• 批准号: 10673513
• 负责人: IGOR R EFIMOV
• 金额: $ 46.67万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10673513
• 关键词: 3-Dimensional; Address; Adopted; Adoption; Adult; Advanced Development; Animal Model; Arrhythmia; Biological Assay; Biomedical Engineering; Blood; Calcium; Cardiac; Cardiac Myocytes; Cardiology; Cardiomyopathies; Cardiovascular system; Cell Differentiation process; Cells; Clinical; Cytoskeleton; Defect; Development; Device or Instrument Development; Devices; Digoxin; Disease; Disease model; Drug Screening; Dyes; Effectiveness; Electrodes; Electronics; Electrophysiology (science); Ensure; Equipment; FLNC gene; Genes; Genetic; Genetic Models; Genotype; Heart; Heart failure; Human; Hydrogels; Image; Isoproterenol; Life; Measurement; Measures; Mechanics; Membrane; Membrane Potentials; Methods; Microelectrodes; Microfabrication; Modeling; Monitor; Morbidity - disease rate; Mutation; Names; Output; Patients; Performance; Pharmacology Study; Pharmacotherapy; Phenotype; Physiological; Positioning Attribute; Process; Quality Control; Research Personnel; Safety; Sarcomeres; Scientist; Shapes; Skin; Sotalol; Specific qualifier value; Sterilization; Structure; Technology; Technology Assessment; Testing; Therapeutic; Time; Tissue Engineering; Tissues; Urine; Validation; Verapamil; Work; arrhythmogenic cardiomyopathy; cardiac tissue engineering; clinically relevant; computerized data processing; culture plates; data acquisition; disease phenotype; drug testing; experience; experimental study; fabrication; flexibility; flexible electronics; graphical user interface; improved; induced pluripotent stem cell; induced pluripotent stem cell derived cardiomyocytes; inherited cardiomyopathy; instrument; manufacture; mortality; multi-electrode arrays; next generation; novel therapeutics; pharmacologic; precision drugs; process improvement; product development; programs; prototype; research and development; research clinical testing; response; sensor; skills; technology validation; therapeutic evaluation; tissue culture; tool; two-dimensional; voltage

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.

项目摘要/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，并在这些疾病中造成严重的发病率和死亡率。新患者/新基因- 针对不同形式的心肌病的特定疗法正在开发和临床测试中，尽管这种疗法的规模很小 动物模型不能充分捕捉临床相关的患者疾病表型和基因类型。病人- 特定的人类诱导多能干细胞(HiPSCs)可以从容易获得的成人细胞(例如， 血液、皮肤或尿液细胞)，并在心脏样细胞、心肌细胞(hiPSC-CMS)中分化，提供 在人类心肌细胞背景下评估新治疗方法的机会。而HiPSC-CM模型是 这些细胞很有希望用于疾病建模和药物测试，但它们还不成熟，通常会产生非生理性的 产出。工程化心脏组织(EHTS)可以通过将HiPSC-CMS浇注到水凝胶基质中来创建， 由柔性柱支撑，在盘子中创建组织状结构。EHTS提高了细胞成熟度，并提供 获得生理输出，如收缩力量。EHTS中的心律失常建模已证明 具有挑战性，只有通过侵入性的终端实验才能完成，这些实验需要特殊的染料和 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，使收缩能力和磁场潜力可以作为电和机械活动来评估 无限期的同时和非侵入性的。该平台将使个性化药物成为可能 测试就像基因纠正策略，目前正在开发中。我们开发了一个原型 采用这种技术的装置，我们称之为机电监测EHTS(EmEHTs)。我们 演示了emEHTs在传感器阵列上捕捉组织应变和场势的功能。 我们建议通过表征器件从制造到长期的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 质量和产量。为了实现更广泛的适用性，将为以下应用开发图形用户界面 收集和分析设备输出。我们将用先进的成像技术进一步验证这项技术 EMEHTs的钙离子和膜电压的表征。对两者进行评估的药理学研究 将进行收缩性和致心律失常的测试，以证明药物测试和筛选的潜力。 先前从患者体内收集并重新编程的HiPSCs，这些患者的基因突变已知高度依赖于 将使用emEHTs来评估致心律失常的可能性，以展示模拟遗传形式的潜力 心肌病。该项目将由一个由心血管研究人员和 拥有临床心脏病学、心律失常建模、心血管遗传学和 微细加工。该项目的成功完成将带来一个经过充分验证的emEHT工作平台 供在HiPSC培养和分化方法方面有经验的科学家使用。