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 心力衰竭和心律不齐是心肌病的遗传形式的主要临床表现，这 可能是威胁生命的，并在这些疾病中引起明显的发病率和死亡率。新患者/基因型 - 特定的疗法正在开发和临床测试对不同形式的心肌病，但是很小 动物模型不足以捕获临床相关的患者疾病表型和基因型。病人- 特定的人类诱导的多能干细胞（HIPSC）可以是由容易获得的成年细胞创建的（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 在人类心肌细胞环境中评估新疗法的机会。而HIPSC-CM型号是 疾病建模和药物测试的有希望的细胞不成熟，通常会产生非生理学 输出。可以通过将HIPSC-CMS施放到水凝胶矩阵中，可以创建工程心脏组织（EHT） 由灵活的柱子支撑，以在菜肴中创建类似组织的结构。 EHT提高细胞成熟度并提供 获得生理输出，例如收缩力。 EHT中的心律失常建模已证明更多 具有挑战性，只能通过需要专门染料和 成像设备。该项目旨在优化灵活的电子技术在EHT中的应用 培养条件，以便可以将收缩力和现场电位评估为电气和机械活性 同时且无创的时间段。该平台将启用个性化药物 当前正在开发的遗传校正策略等测试。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对EHTS（EMEHTS）的设备。我们 目前证明EMEHTS在传感器阵列上捕获组织应变和场电位的功能。 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，采用标准产品开发质量改进过程以确保设备一致 质量和输出。为了允许更广泛的可采用性，将为图形用户界面开发 收集和分析设备输出。我们将通过高级成像进一步验证这项技术 钙和膜电压的表征。评估两者的药理学研究 将进行收缩和心律失常，以证明药物测试和筛查的潜力。 先前收集和重编程的HIPSC来自具有高度的基因突变的患者 心律失常将使用EMEHT进行评估，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，心血管遗传学和心律失常技能的生物工程师 微加工。该项目的成功完成将导致验证良好的工作EMEHT平台 科学家在HIPSC文化和分化方法中使用经验。