CAREER: Wireless Flexible Micro-sensors to Monitor Cardiac Phenotypes in Zebrafish Models of Heart Regeneration

在发达国家，心脏疾病是导致死亡的主要原因，原因是由于血凝块或其他形式的阻塞导致的血流损失导致的心室心肌（心壁）损伤不能充分替换。这种失败归因于成年哺乳动物心室心肌细胞（CMs =心肌细胞）分裂和再生失去的心肌的能力有限。相比之下，斑马鱼的心脏在20%心室切除后完全再生，因此为心脏再生研究提供了一个遗传上可处理的模型系统。虽然斑马鱼心脏已被广泛评估，使用免疫组织化学和DNA和蛋白质分析来研究不同信号通路在心脏发育和再生中的作用，这被认为与开发哺乳动物再生疗法有关，但目前的方法无法阐明同一样本随时间的过程（即再生）的进展。该项目的目标是建立在PI在镇静动物中获得高质量（有利的高信噪比）心电图（ECG）信号的能力基础上，为自由游动的斑马鱼心脏再生模型提供革命性的基于聚合物的无线设备，用于长期获取固有ECG。更广泛的社会利益包括：1)实现新的心脏治疗；2)降低药物筛选的成本和时间；3)为患者和健康人群提供大量舒适的贴片式医疗可穿戴设备铺平道路。研究与教育活动的整合旨在帮助缩小工程教育与生物医学研究的差距，包括努力1)激励和培养来自不同学科和水平的学生接受高等教育和生物工程事业；2)通过教师直接或间接地培养幼稚园学生，拓宽他们在生物工程和医学研究中应用的最新高科技知识；3)将严谨研究中使用的尖端技术转化为直接支持社会的现实世界设备；4)聘请工程、科学和医学多学科人才，解决未满足的生物工程挑战；5)建立国际合作，交流创新，并解决全球问题，如水质。由于成年心肌细胞（CMs）的分裂和增殖能力有限，哺乳动物的心脏在缺血梗死后再生心肌的能力有限，而斑马鱼（Danio rerio）的心脏在切除20%心室后完全再生。该项目的中心主题在于应用柔性和可拉伸微电子和无线电力传输来进行自由游动的斑马鱼模型的心脏再生研究。该项目包括三个里程碑：1)开发柔性和可拉伸的微电极阵列（MEA）膜，用于在斑马鱼心脏再生模型中长期记录心电图（ECG）：四个直径约200微米的工作电极将放置在心脏附近，一个参考电极将放置在身体上。2)设计并实现一种无线系统（“ECG Jacket”），包括无线供电（电感耦合或超声）和数据通信，用于在生物研究下对圆形水箱中自由游动的鱼（同时监测多条鱼）进行远程连续心电监测。3)利用该系统阐明特定基因在斑马鱼（截肢或低温方法诱导损伤的多突变样模型）心肌再生中的作用，并具有潜在的人类翻译潜力，并展示翻译潜力1)药物筛选(斑马鱼将使用影响心脏活动的知名药物治疗；例如胺碘酮和维拉帕兰)和ii)在家庭环境中使用舒适（灵活和可拉伸）和不显眼的贴片设备（例如ECG， EEG， EOG， EMG和血压）进行人体生理监测。所开发的技术将能够对特定心肌部位进行长期（长达2个月）的心电图记录，从而在不使用镇静的情况下独特地确定所调查区域的整体功能。哺乳动物心脏几乎没有再生能力的传统观点现在受到最近动物和人类研究的质疑，在这些研究中，新的CMs可能来自现有的CMs和祖细胞或干细胞。在斑马鱼研究中发现了特定基因在心脏再生中的作用，这将为激活人类心脏有限的再生能力提供方法，使人们对潜在的心脏治疗方法感到乐观。

PI: Cao, HungProposal #: 1652818Heart diseases are the leading cause of death in the developed world due to failure to adequately replace lost ventricular myocardium (heart wall) damaged by loss of blood flow due to blood clots or other forms of obstruction. This failure is attributed to the limited ability of adult mammalian ventricular cardiomyocytes (CMs = heart muscle cells) to divide and regenerate the lost myocardium. In contrast, zebrafish hearts fully regenerate after 20% ventricular removal and thus provide a genetically tractable model system for heart regeneration investigations. While the zebrafish heart has been extensively assessed using immunohistochemistry and DNA and protein analyses to study the roles of different signaling pathways in cardiac development and regeneration, which is thought to have relevance for developing mammalian regeneration therapies, current approaches cannot elucidate the progress of the process (i.e., regeneration) of the same samples over time. The objective of this project, which builds on the PI's demonstrated ability to obtain quality (favorable high signal to noise ratio) electrocardiogram (ECG) signals in sedated animals, is to provide revolutionary polymer-based wireless devices for long-term acquisition of intrinsic ECG in freely swimming zebrafish models of heart regeneration. Broader society interests include the promise 1) to enable novel cardiac therapy; 2) to reduce cost and time of drug screening; and 3) to pave the way for numerous comfortable patch-based healthcare wearables for both patients and healthy populations. Integration of research with education activities are designed to help close the gap of engineering education and biomedical research, including efforts to 1) inspire and train students from different disciplinary and levels to follow higher education and a bioengineering career; 2) to nurture K-12 students directly and indirectly through their teachers by broadening knowledge in state-of-the-art hi-technology applied in bioengineering and medical research; 3) to translate cutting-edge technologies used in rigorous research to real world devices directly supporting society; 4) to engage multidisciplinary personnel from engineering, science and medicine to tackle unmet bioengineering challenge; and 5) to establish international collaborations to exchange innovations as well as to address global issues such as water quality.Unlike mammalian hearts, which have limited ability to regenerate myocardium after ischemia induced infarct due to the limited capacity of adult cardiomyocytes (CMs) to divide and proliferate, zebrafish (Danio rerio) hearts fully regenerate after 20% ventricular resection. The central theme of this project lies in the applications of flexible and stretchable microelectronics and wireless power transfer to carry out heart regeneration studies in freely swimming zebrafish models. The project involves three milestones: 1) Development of flexible and stretchable micro-electrode array (MEA) membranes for long-term recording of electrocardiogram (ECG) in zebrafish models of heart regeneration: Four working electrodes approximately 200 um in diameter will be placed near the heart and a reference electrode will reside on the body. 2) Design and implementation of a wireless system (an "ECG Jacket") including wireless powering (inductive coupling or ultrasound) and data communication for remote and continuous electrocardiac monitoring of freely swimming fish (multiple fish monitored simultaneously) in a circular tank under biological investigations. 3) Deploying the system to elucidate the roles of specific genes towards myocardium regeneration in zebrafish (multiple mutant-like models with injuries induced by either amputation or a cryogenic approach) with potential translations to humans and to demonstrate the translational potentials for i) drug screening (zebrafish will be treated with well-known drugs that affect cardiac activity, e.g. amiodarone and verapamiland) and ii) physiological monitoring in humans with comfortable (flexible and stretchable) and unobtrusive patch-based devices (e.g., ECG, EEG, EOG, EMG, and blood pressure) in the home setting. The technologies developed will enable long-term (up to 2 months) ECG recordings of specific myocardial sites and thus uniquely determine the overall functionality of the area under investigation without effects of sedation. The conventional view of mammalian hearts as having virtually no regenerative capacity is now questioned by recent animal and human studies, in which new CMs may arise from existing CMs and progenitor or stem cells. The discovery of specific genes' roles towards heart regeneration in zebrafish studies would suggest methods to activate limited regenerative capacity in the human heart, garnering optimism about potential cardiac therapies.