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.

项目概要/摘要 心力衰竭和心律失常是遗传型心肌病的主要临床表现， 这些疾病可能危及生命并导致显着的发病率和死亡率。新患者/基因型- 针对不同形式的心肌病（无论规模大小）的特定疗法正在开发和临床测试中 动物模型不足以捕捉临床相关的患者疾病表型和基因型。病人- 特定的人类诱导多能干细胞 (hiPSC) 可以从现成的成体细胞（例如， 血液、皮肤或尿液细胞）并分化为类心脏细胞、心肌细胞（hiPSC-CM），提供 在人类心肌细胞背景下评估新疗法的机会。虽然 hiPSC-CM 模型是 这些细胞在疾病建模和药物测试方面很有前景，但它们还不成熟，经常产生非生理性的结果。 输出。工程心脏组织 (EHT) 可以通过将 hiPSC-CM 注入水凝胶基质中来创建， 由柔性柱支撑，在培养皿中形成组织状结构。 EHT 可提高细胞成熟度并提供 获得生理输出，如收缩力。 EHT 中的心律失常建模已被证明更多 具有挑战性，并且只能通过需要特殊染料和的侵入性最终实验来完成 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，以便可以将收缩性和场电位评估为电气和机械活动 同时且无创地无限期地进行。该平台将使个性化药物成为可能 诸如基因校正策略之类的测试目前正在开发中。我们开发了一个原型 采用该技术的设备，我们将其命名为机电监控 EHT (emEHT)。我们 目前演示了 emEHT 的功能是捕获组织应变和传感器阵列上的场电位。 我们建议通过表征从制造到长期的器件功能来进一步验证该技术 文化，采用标准的产品开发质量改进流程来确保设备的一致性 质量和产出。为了实现更广泛的采用，将开发一个图形用户界面 收集和分析设备输出。我们将通过先进的成像进一步验证这项技术 emEHT 的钙和膜电压的表征。评估两者的药理学研究 将进行收缩性和心律失常性以证明药物测试和筛选的潜力。 先前从携带已知高度突变基因的患者中收集并重新编程的 hiPSC 将使用 emEHT 评估致心律失常性，以证明对心律失常的遗传形式进行建模的潜力 心肌病。该项目将由心血管研究人员和跨学科团队共同领导 具有临床心脏病学、心律失常建模、心血管遗传学等方面技能的生物工程师 微加工。该项目的成功完成将产生一个经过充分验证的工作 emEHT 平台 供具有 hiPSC 培养和分化方法经验的科学家使用。