Optically Promoting Cardiac Maturation Using Engineered Peptides

使用工程肽光学促进心脏成熟

• 批准号: 10628281
• 负责人: Herdeline Ann Mallari Ardoña
• 金额: $ 50.12万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10628281
• 关键词: Action Potentials; Address; Adult; Animal Model; Anisotropy; Architecture; Automobile Driving; Behavior; Binding; Biophysics; Calcium; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Cardiovascular system; Cell Culture Techniques; Cell Survival; Cells; Charge; Chemical Stimulation; Cues; Drug Screening; Electric Stimulation; Electrodes; Engineering; Ensure; Epitopes; Film; Frequencies; Functional disorder; Gene Expression; Generations; Goals; Heart Diseases; Heart Injuries; Heating; Human; In Vitro; Investigation; Light; Measurement; Measures; Mechanics; Mediating; Metabolism; Methods; Microelectrodes; Modeling; Monitor; Muscle Cells; Nanostructures; Neonatal; Optics; Pattern; Peptide Library; Peptides; Phenotype; Polymers; Process; Protocols documentation; Rattus; Reaction; Reporting; Research; Resolution; Risk Assessment; Risk Reduction; Side; Signal Transduction; Source; Stimulus; Stress; Structure; Surface; Technology; Testing; Therapeutic; Tissues; Toxicology; Transducers; Transfection; Ventricular; biomaterial interface; cardiac tissue engineering; chromophore; combinatorial; conditioning; design; disease mechanisms study; drug discovery; extracellular; genetically modified cells; high throughput screening; human stem cells; improved; indexing; induced pluripotent stem cell; infection risk; innovation; instrumentation; irradiation; minimally invasive; monomer; non-genetic; novel strategies; optogenetics; personalized medicine; regenerative therapy; response; scale up; spatiotemporal; species difference; stem; stem cell based approach; stem cells; success; temporal measurement; wireless

PROJECT SUMMARY The promise of human stem cell-derived cardiomyocytes (hSC-CMs) opens doors towards the feasibility of personalized medicine against cardiac diseases and for performing more accurate drug discovery studies. Moreover, hSC-CMs overcome the issue of species differences when using animal models for high throughput screening studies. However, one of the bottlenecks for scaling up the use of hSC-CMs is their ability to accurately reflect the native structure and function of adult human cardiomyocytes. Current efforts to address this critical challenge involve maturation protocols that use biophysical cues such as electrical stimulation or substrate- induced tissue alignment. Methods for electrical stimulation often utilize electrode contacts for field stimulation, bulky instrumentation for combining electrical stimuli delivery with mechanical or sustained chemical stimulation, or genetically modifying cells to be light-responsive. Although we have seen successes through these induction and stimulation approaches, the field would benefit from a stimulation approach with minimal culture contact to reduce risk of infection during long-term cultures, as well as a light-based approach with higher spatiotemporal resolution than electrode-based stimulation. Here, we propose a new paradigm for stimulating hSC-CMs towards maturation by interfacing these cells with peptide-based substrates that can induce tissue anisotropy and are engineered to convert light to stimulatory cues. Our team will develop peptides functionalized with chromophore units and cell-binding epitopes as materials that can be used for light-based stimulation of hSC- CMs, in combination with induction of tissue alignment, towards maturation. The long-term goal of this project is to establish light stimulation via engineered peptides as a viable method to stimulate cardiomyocytes and promote hSC-CM maturation in an electrodeless and non-genetic manner. We hypothesize that transient charging and other associated light-induced processes at the cardiomyocyte-biomaterial interface can influence extracellular potential, resulting in the photostimulation of hSC-CMs towards maturation. Our rationale for proposing a materials-based approach for stimulating hSC-CMs stems from previous reports of conjugated polymers being used as a photoactive substrate for triggering action potentials of other excitable cells. To test our hypothesis, we propose the following specific aims: (1) establishing design parameters for engineered peptide substrates with optimal photostimulation efficiency; (2) test the cellular- and tissue-level impact of peptide-mediated photostimulation in combination with anisotropic cues; and (3) elucidate the effect of the proposed photostimulation method, along with anisotropy cues, on hSC-CM maturation. By establishing the design rules for the proposed photoexcitable peptides for eliciting combinatorial cues to stimulate hSC-CMs and ensure their capability to excite cardiac cells, this innovative approach offers a new strategy for a “wireless” stimulation of cardiac tissues towards maturation, and can therefore significantly contribute towards addressing the grand challenge of immaturity of stem cell-derived cardiomyocytes.

项目摘要 人类干细胞衍生的心肌细胞（HSC-CMS）的承诺为可行性打开门 针对心脏病的个性化医学，并进行更准确的药物发现研究。 此外，HSC-CMS在使用动物模型进行高通量时克服了物种差异的问题 筛查研究。但是，扩大HSC-CMS使用的瓶颈之一是它们准确的能力 反映成人人类心肌细胞的天然结构和功能。当前解决这一关键的努力 挑战涉及使用生物物理提示（例如电刺激或底物）的成熟方案 诱导的组织比对。电刺激的方法通常利用电极触点进行场刺激， 笨重的仪器，用于将电刺激递送与机械或持续化学刺激相结合的仪器， 或基因修饰细胞具有光响应性。尽管我们通过这些归纳而看到了成功 和刺激方法，该领域将受益于与最小培养接触的刺激方法 降低长期培养期间感染的风险，以及具有较高时空的光基方法 分辨率比基于电极的刺激。在这里，我们提出了一个用于刺激HSC-CMS的新范式 通过将这些细胞与基于肽的底物连接到可以诱导组织各向异性和的成熟 经过精心设计，以将光转换为刺激提示。我们的团队将开发功能化的肽 发色团单元和细胞结合表位作为可用于HSC-基于光基模拟的材料 CMS与组织比对的诱导结合成熟。这个项目的长期目标是 通过工程辣椒作为刺激心肌细胞和 以无电和非遗传方式促进HSC-CM成熟。我们假设那个瞬态 心肌细胞生物材料界面处的充电和其他相关的光引起的过程会影响 细胞外电势，导致HSC-CM的光刺激对成熟。我们的理由 提出一种基于材料的方法来刺激先前报告的报告中的HSC-CMS步骤 聚合物被用作光活性底物，用于触发其他令人兴奋的细胞的作用电位。测试 我们的假设，我们提出以下特定目的：（1）建立工程设计的设计参数 具有最佳光刺激效率的肽底物； （2）测试细胞和组织水平的影响 肽介导的光刺激与各向异性提示结合； （3）阐明 提出的光刺激方法以及各向异性提示在HSC-CM成熟方面。通过建立 提议的可启用Pepperides的设计规则，用于引发组合提示，以刺激HSC-CMS和 确保它们激发心脏细胞的能力，这种创新的方法为“无线”提供了新的策略 刺激心脏组织成熟，因此可以显着有助于解决 干细胞衍生的心肌细胞不成熟的巨大挑战。