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

使用基于亲和力的工程生物材料调节组织修复中的蛋白质活性

• 批准号: 10655635
• 负责人: Marian Hirushika Hettiaratchi
• 金额: $ 36.88万
• 依托单位: UNIVERSITY OF OREGON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10655635
• 关键词: Affect; Affinity; Area; Binding; Biocompatible Materials; Bone Injury; Complex; Computer Models; Development; Directed Molecular Evolution; Engineering; Implant; Injury; Knowledge; Libraries; Modeling; Outcome; Process; Protein Engineering; Protein Inhibition; Proteins; Regenerative Medicine; Role; Signaling Protein; Site; Societies; Specificity; Surface; Tertiary Protein Structure; Testing; Tissues; Work; Yeasts; delivery vehicle; design; flexibility; healing; improved; injured; innovation; insight; interest; novel; protein complex; receptor; receptor binding; regenerative; response; spatiotemporal; tissue regeneration; tissue repair; tool; wound healing

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.

项目总结 协调的蛋白质信号是协调体内许多功能所必需的。在组织修复过程中， 蛋白质在损伤部位的时空呈现影响下游细胞的蛋白质-受体结合 反应和总体治愈结果。许多生物材料已经被设计成能够输送蛋白质来治疗 受伤的组织。然而，很少有生物材料能够独立地控制从一种生物材料中输送多种蛋白质 单一材料，限制了它们传递与自然伤口愈合有关的多种蛋白质的效用 卡斯卡德。控制蛋白质输送的一种策略是使用基于亲和力的生物材料，这种材料使用非 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力的生物材料来 通过确定复杂蛋白质组合的时间和局部呈现方式来增强组织修复 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是如何- 物质亲和力相互作用影响蛋白质的释放和活性，调节复杂的愈合反应，以及 询问蛋白质呈现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输送生物材料的发展：1)蛋白质-材料亲和力如何 相互作用影响蛋白质的释放和活性？2)基于生物材料的如何控制蛋白质的呈现 会影响组织修复吗？我们的创新方法包括设计新的蛋白质-材料亲和力相互作用 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的蛋白质结构域 (即亲和体)与目的蛋白结合，具有高度的特异性和广泛的亲和力。计算型 建模将被用来设计与蛋白质不同区域相互作用的仿生体以抑制或维持 蛋白质-受体结合。由此产生的大量亲和抗体将使我们能够确定蛋白质是如何- 物质亲和力的相互作用在不同的时间尺度上影响蛋白质的释放和活性。亲和体将会是 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库 生物材料，我们将系统地研究多个蛋白质的时间呈现如何影响速率 和组织修复的质量。我们将植入生物材料，以恢复愈合反应的关键方面1) 使用中等亲和力的亲和体将外源蛋白持续输送到损伤部位和2) 使用高亲和力亲和体将内源性蛋白隔离在损伤部位，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，但我们将首先在骨损伤模型中测试它， 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表达 治愈级联，我们将获得对许多蛋白质的作用的新见解，这些蛋白质管理组织修复并创建 新型高度模块化的生物材料，可以量身定做，协调细胞反应，治疗多发性 受伤的类型。我们的方法将通过创建一个变革性的 新的再生医学策略具有显著改善组织修复的潜力。