Modulating Protein Activity in Tissue Repair using Engineered Affinity-based Biomaterials

项目摘要 协调蛋白质信号传导是协调体内许多功能所必需的。在组织修复过程中， 损伤部位蛋白质的时空呈递影响蛋白质-受体结合，下游细胞 反应和整体愈合结果。许多生物材料已经被设计成递送蛋白质以治疗 受伤的组织然而，很少有生物材料可以独立地控制多种蛋白质从微球的递送。 单一材料，限制了它们用于递送参与自然伤口愈合的多种蛋白质的效用 级联。控制蛋白质递送的一种策略是使用基于亲和性的生物材料，其采用非生物相容性材料。 蛋白质和材料之间的共价亲和力相互作用。我的实验室正在开发基于亲和性的生物材料， 通过确定蛋白质的复杂组合的时间和局部呈现， 影响再生过程。我们的目标是开发新的生物材料工具来了解蛋白质- 材料亲和相互作用影响蛋白质释放和活性，调节复杂的愈合反应， 询问蛋白质呈递在组织修复中的作用。我们将解决两个关键的知识差距， 阻碍了有效的蛋白质递送生物材料的发展：1）蛋白质-材料亲和力如何 相互作用影响蛋白质释放和活性？2)基于生物材料的蛋白质呈递控制 影响组织修复吗我们的创新方法涉及利用生物技术设计新的蛋白质-材料亲和相互作用， 定向进化和合理的蛋白质设计。酵母表面展示将用于进化小蛋白质结构域 (i.e.,抗体），其以高特异性和广泛的亲和力结合感兴趣的蛋白。计算 模型将用于设计与蛋白质的不同区域相互作用的抗体，以抑制或维持 蛋白质受体结合由此产生的大量的蛋白质抗体将使我们能够确定蛋白质- 材料亲和性相互作用在不同的时间尺度上影响蛋白质的释放和活性。Affibodies将是 结合到生物材料上以调节蛋白质释放和细胞反应。使用我们的基于亲和力的 生物材料，我们将系统地研究如何多个蛋白质的时间介绍影响率 和组织修复的质量。我们将植入生物材料，通过以下方式恢复愈合反应的关键方面：1） 使用中等亲和性抗体以向损伤部位提供外源蛋白质的持续递送，和2） 使用高亲和性抗体隔离损伤部位内的内源性蛋白质，并增强、维持或 抑制蛋白质活性。虽然我们的方法是灵活的和组织不可知的，我们将首先在骨损伤模型中进行测试， 这是我实验室的核心专业领域通过复制伤口的复杂蛋白质呈现 愈合级联，我们将获得新的见解，许多蛋白质的作用，管理组织修复和创造一个 新型高度模块化生物材料，可定制以协调细胞反应，以治疗多种 受伤的类型。我们的方法将通过创造一个变革性的社会， 新的再生医学策略，具有显着改善组织修复的潜力。

PROJECT SUMMARY Coordinated protein signaling is required to orchestrate many functions in the body. During tissue repair, the spatiotemporal presentation of proteins in the injury site affects protein-receptor binding, downstream cellular responses, and overall healing outcomes. Many biomaterials have been designed to deliver proteins to treat injured tissues. However, few biomaterials can independently control the delivery of multiple proteins from a single material, limiting their utility for delivering numerous proteins involved in the natural wound healing cascade. One strategy to control protein delivery is the use of affinity-based biomaterials, which employ non- covalent affinity interactions between proteins and materials. My lab is developing affinity-based biomaterials to enhance tissue repair by determining how the timing and local presentation of complex combinations of proteins affect regenerative processes. Our objective is to develop new biomaterial tools to understand how protein- material affinity interactions impact protein release and activity, modulate complex healing responses, and interrogate the role of protein presentation in tissue repair. We will tackle two critical knowledge gaps that have hindered the development of effective biomaterials for protein delivery: 1) How do protein-material affinity interactions affect protein release and activity? 2) How does biomaterial-based control over protein presentation affect tissue repair? Our innovative approach involves engineering new protein-material affinity interactions using directed evolution and rational protein design. Yeast surface display will be used to evolve small protein domains (i.e., affibodies) that bind to proteins of interest with high specificity and a wide range of affinities. Computational modeling will be used to design affibodies that interact with different areas of the protein to inhibit or maintain protein-receptor binding. The resulting expansive array of affibodies will allow us to determine how protein- material affinity interactions affect protein release and activity over different timescales. Affibodies will be conjugated onto biomaterials to tune protein release and cellular responses. Using our library of affinity-based biomaterials, we will systematically investigate how the temporal presentation of multiple proteins affects the rate and quality of tissue repair. We will implant biomaterials to restore key aspects of the healing response by 1) using moderate affinity affibodies to provide sustained delivery of exogenous proteins to the injury site and 2) using high affinity affibodies to sequester endogenous proteins within the injury site and enhance, maintain, or inhibit protein activity. While our approach is flexible and tissue-agnostic, we will first test it in a bone injury model, which is a central area of expertise in my lab. By replicating the complex protein presentation of the wound healing cascade, we will gain new insights into the roles of many proteins that govern tissue repair and create a new class of highly modular biomaterials that can be tailored to orchestrate cellular responses to treat multiple types of injuries. Our approach will have an immediate benefit to society through the creation of a transformative new regenerative medicine strategy with the potential to significantly improve tissue repair.