Optically Promoting Cardiac Maturation Using Engineered Peptides

项目总结 人类干细胞来源的心肌细胞(HSC-CMS)的前景为 心脏病个体化用药的可行性及实施更准确用药的可行性 发现性研究。此外，HSC-CMS克服了使用动物时物种差异的问题 用于高通量筛查研究的模型。然而，扩大使用的瓶颈之一是 HSC-CMS是一种能够准确反映成人天然结构和功能的能力 心肌细胞。目前解决这一关键挑战的努力涉及使用 生物物理线索，如电刺激和机械刺激。这些方法通常使用电极 用于现场刺激的触点，用于机械或持续化学刺激的笨重仪器，或 对细胞进行基因改造，使其对光有反应。尽管我们已经通过这些取得了成功 诱导和刺激方法，领域将受益于以最少的刺激方法 文化接触，以减少长期培养期间的感染风险，以及基于光线的方法 比基于电极的刺激具有更高的时空分辨率。在这里，我们提出了一个新的 通过多肽结合刺激HSC-CMS走向成熟的范例 将光转换为电信号的基片。我们的团队将开发多肽 以生色团单元和细胞结合表位为材料设计而成，可用于 HSC-CMS成熟过程中的光电条件作用。这个项目的长期目标是 通过工程多肽建立光电调节作为一种可行的电刺激方法 并以无电极和非遗传方式促进HSC-CM成熟， 比野外刺激更高的时空分辨率。我们假设瞬变充电和其他 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-CMS的光电刺激走向成熟。我们的理由是 提出一种基于材料的方法来刺激HSC-CMS源于先前关于 共轭聚合物被用作触发其他生物动作电位的光活性底物 可兴奋的细胞。为了验证我们的假设，我们提出了以下具体目标：(1)建立设计 具有最佳光刺激效率的多肽纳米组装体的参数；(2)测试细胞和 多肽介导的光刺激对组织水平的影响；以及(3)阐明所提出的 光电调节方法对HSC-CM成熟的影响。通过为 用于刺激HSC-CMS并确保其局部兴奋能力的光可激肽 心脏细胞，这种创新的方法提供了一种“无线”刺激心脏组织的新策略。 并可为应对干细胞不成熟这一重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