Optically Promoting Cardiac Maturation Using Engineered Peptides

项目概要 人类干细胞衍生的心肌细胞（hSC-CM）的前景为实现这一目标打开了大门 针对心脏病的个性化医疗和执行更准确药物的可行性 发现研究。此外，hSC-CM克服了使用动物时的物种差异问题 高通量筛选研究的模型。然而，扩大使用规模的瓶颈之一 hSC-CM 能够准确反映成年人的天然结构和功能 心肌细胞。当前解决这一关键挑战的努力涉及使用成熟协议 生物物理线索，例如电刺激和机械刺激。这些方法通常利用电极 用于场刺激的触点、用于机械或持续化学刺激的大型仪器，或 对细胞进行基因改造，使其具有光响应能力。尽管我们已经通过这些看到了成功 诱导和刺激方法，该领域将受益于刺激方法，以最小的 培养接触以降低长期培养期间感染的风险，以及基于光的方法 与基于电极的刺激相比，具有更高的时空分辨率。在这里，我们提出一个新的 通过将这些细胞与基于肽的接口来刺激 hSC-CM 成熟的范例 设计用于将光转换为电信号的基板。我们的团队将开发肽 以发色团和细胞结合表位作为材料进行工程设计，可用于 hSC-CM 的光电调节使其成熟。该项目的长期目标是 通过工程肽建立光电调节作为电刺激的可行方法 心肌细胞并以无电极和非遗传方式促进 hSC-CM 成熟， 比场刺激具有更高的时空分辨率。我们假设瞬态充电和其他 心肌细胞-生物材料界面的相关光诱导过程可以影响细胞外 电位，导致 hSC-CM 的光电刺激使其成熟。我们的理由 提出一种基于材料的刺激 hSC-CM 的方法源于之前的报告 共轭聚合物用作光活性底物，用于触发其他物质的动作电位 兴奋细胞。为了检验我们的假设，我们提出以下具体目标：（1）建立设计 具有最佳光刺激效率的肽纳米组装体的参数； (2) 测试蜂窝和 肽介导的光刺激对组织水平的影响； (3) 阐明建议的效果 光电调节方法对 hSC-CM 成熟的影响。通过建立设计规则 提出了用于刺激 hSC-CM 并确保其局部兴奋能力的光激发肽 心脏细胞，这种创新方法为心脏组织的“无线”刺激提供了新策略 并可以为解决干细胞不成熟的巨大挑战做出重大贡献。 1

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 and mechanical stimulation. These methods often utilize electrode contacts for field stimulation, bulky instrumentation for 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 are engineered to convert light to electrical cues. Our team will develop peptides engineered with chromophore units and cell-binding epitopes as materials that can be used for photoelectrical conditioning of hSC-CMs towards maturation. The long-term goal of this project is to establish photoelectrical conditioning via engineered peptides as a viable method to electrically stimulate cardiomyocytes and promote hSC-CM maturation in an electrodeless and non-genetic manner, with higher spatiotemporal resolution than field stimulation. We hypothesize that transient charging and other associated light-induced processes at the cardiomyocyte-biomaterial interface can influence extracellular potential, resulting in the photoelectrical stimulation 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 peptide nanoassemblies with optimal photostimulation efficiency; (2) test the cellular- and tissue-level impact of peptide-mediated photostimulation; and (3) elucidate the effect of the proposed photoelectrical conditioning method on hSC-CM maturation. By establishing the design rules for the proposed photoexcitable peptides for stimulating hSC-CMs and ensuring their capability to locally excite cardiac cells, this innovative approach offers a new strategy for a “wireless” stimulation of cardiac tissues and can significantly contribute towards addressing the grand challenge of stem cell immaturity. 1