LightKick: Novel bioengineering system for activity-dependent acceleration of functional maturation of human stem cell-derived cardiomyocytes

LightKick：新型生物工程系统，可加速人类干细胞来源的心肌细胞的功能成熟

• 批准号: 10081239
• 负责人: ALEX SAVTCHENKO
• 金额: $ 32.5万
• 依托单位: NANOTOOLS BIOSCIENCE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10081239
• 关键词: Academia; Acceleration; Address; Adult; Area; Biological Assay; Biomedical Engineering; Biotechnology; Cardiac; Cardiac Myocytes; Cardiotoxicity; Cardiovascular Agents; Cardiovascular Diseases; Cardiovascular system; Cell Culture Techniques; Cell Survival; Cells; Clinical; Coculture Techniques; Communities; Complex; Computer software; Consensus; Cues; Cultured Cells; Development; Disease; Dose; Drug Industry; Elasticity; Electrodes; Embryo; Engineering; Fibroblasts; G-substrate; Genetic; Heart; Heart Diseases; Human; Hybrids; Incubators; Lead; Light; Lighting; Mediating; Modeling; Modification; Monitor; Optics; Outcome; Pain; Patients; Pattern; Phenotype; Physiological; Physiology; Play; Population; Problem Solving; Process; Property; Protocols documentation; Regenerative Medicine; Replacement Therapy; Reproducibility; Research; Research Personnel; Scientist; Signal Transduction; Stimulus; Structure; System; Technology; Testing; Thick; Time; Tissues; Translating; Wireless Technology; base; biomaterial compatibility; biophysical properties; cell type; clinical application; cost effective; culture plates; design; drug development; drug discovery; electric field; experimental study; flexibility; graphene; human stem cells; improved; induced pluripotent stem cell; innovation; insight; mortality; novel; optogenetics; personalized therapeutic; polydimethylsiloxane; precision medicine; programs; prototype; public health relevance; regenerative; remote control; scaffold; screening; stem cells; success; tool; user-friendly

ABSTRACT Human iPSC-derived cardiomyocytes offer infinite possibilities for cardiovascular drug discovery and regenerative medicine, but only if they reflect the physiology of an adult human heart. Current protocols produce cardiomyocytes with embryonic rather than adult profiles, severely limiting their utility for the adult population. Recent studies suggest that bioengineering approaches aimed at re-creating environmental cues in a dish can lead to marked improvements in cardiomyocytes’ maturation profiles. New technological tools are required to translate these findings into the practice. To enable scientists in academia and biotech/pharmaceutical industry to generate more mature hiPSC-derived cardiomyocytes on demand, we propose to design, engineer, fabricate, and validate a novel bioengineering hardware system that will optically control the pacing of hiPSC-derived cardiomyocytes over extended periods of time to drive their maturation toward the adult phenotype in an activity-dependent manner. The proposed maturation-promoting system will utilize our proprietary graphene-mediated optical stimulation technology that allows fast, reversible, and physiologically relevant activation of cells. Our technology has several critical advantages, as (1) it requires neither genetic modifications (vs. optogenetics) nor wires/electrodes (vs. electric field stimulation), (2) it offers an uncharted flexibility for tunable light- controlled cell stimulation protocols, and (3) it is compliant with high-throughput and scalability requirements. The components of the proposed system (LightKick) will include graphene-coated microplates (G-plates), a multi-LED unit to illuminate cells cultured in G-plates inside a cell incubator, and an external multiparametric programmable controller to define illumination parameters. G-plates have already been developed and validated for optical stimulation and all-optical assays. We will further optimize G-plates in terms of the substrate rigidity and micropatterning. The development of two other components, together comprising an optical stimulation module, will represent the main engineering efforts of the proposed project. To tailor this module for long-term optical stimulation, we will take into consideration recently developed light emitters, optoelectronic properties of G-plates, biophysical properties of cardiomyocytes, and long-term cell culturing conditions. Extensive technical capabilities of our optical stimulation module will offer (1) the scalability (by delivering stimulation signals in few wells or few plates on demand); (2) wireless monitoring of the module and remote control of optical pacing; and (3) the flexible tunability across the wide range of physiologically-relevant light illumination parameters using a user-friendly interactive GUI software. Importantly, long-term optical stimulation of hiPSC-derived cardiomyocytes enabled by the LightKick would be fully compatible with many environmental cues that might play in the maturation process. The LightKick system will allow stem cell experts (1) to develop efficient and individualized cell stimulation patterns for producing adult-like patient-specific hiPSC-derived cells, and (2) to easily combine dynamic electric environmental cues with other environmental cues, further improving multiparametric maturation-promoting protocols. We anticipate that the LightKick system will dramatically simplify and, thus, accelerate the process of finding fundamental and practical answers to the maturation problem.

摘要 人类iPSC衍生的心肌细胞为心血管药物发现和再生提供了无限可能性 医学，但前提是它们反映成年人心脏的生理机能。目前的方案产生心肌细胞， 胚胎而不是成人的轮廓，严重限制了它们对成人群体的效用。最近的研究表明， 旨在在培养皿中重新创造环境线索的生物工程方法可以显著改善 心肌细胞的成熟概况。需要新的技术工具将这些研究结果转化为实践。 为了使学术界和生物技术/制药行业的科学家能够产生更成熟的hiPSC衍生的 心肌细胞的需求，我们建议设计，工程师，制造和验证一种新的生物工程硬件系统 这将在很长一段时间内光学控制hiPSC衍生的心肌细胞的起搏，以驱动它们的 以活动依赖性方式向成人表型成熟。建议的成熟促进制度将 利用我们专有的石墨烯介导的光学刺激技术， 相关的细胞激活。我们的技术有几个关键的优势，因为（1）它既不需要基因改造， (vs.光遗传学）或导线/电极（相对于电场刺激），（2）它为可调谐光提供了前所未有的灵活性， 受控的细胞刺激方案，和（3）它符合高通量和可扩展性要求。 拟议的系统（LightKick）的组件将包括石墨烯涂层微孔板（G板），一个多LED 用于照射细胞培养箱内G板中培养的细胞的单元，以及外部多参数可编程控制器 以定义照明参数。G板已经开发并验证了光学刺激和全光学 分析。我们将进一步优化G-板在基板刚度和微图案方面。开发两 其他部件一起构成光学刺激模块，将代表本发明的主要工程努力。 拟议项目。为了使该模块适合长期光学刺激，我们将考虑最近开发的 发光体、G板的光电特性、心肌细胞的生物物理特性和长期细胞培养 条件我们的光学刺激模块的广泛技术能力将提供（1）可扩展性（通过提供 根据需要在少数威尔斯或少数板中的刺激信号）;（2）模块的无线监测和 光学起搏;以及（3）在生理相关光照参数的宽范围内的灵活可调性 使用用户友好的交互式GUI软件。 重要的是，通过LightKick实现的对hiPSC衍生的心肌细胞的长期光学刺激将被完全抑制。 与许多可能在成熟过程中起作用的环境线索相一致。LightKick系统将允许股骨柄 细胞专家（1）开发有效和个性化的细胞刺激模式，用于产生成人样的患者特异性 hiPSC衍生的细胞，和（2）容易地将动态电环境线索与其他环境线索结合，进一步 改进多参数促成熟方案。我们预计，LightKick系统将显着 简化，从而加速寻找成熟问题的基本和实际答案的过程。