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.

项目摘要 需要协调的蛋白质信号传导来编排体内许多功能。在组织修复期间， 损伤部位蛋白质的时空表现会影响蛋白质受体结合，下游细胞 反应和整体治愈结果。许多生物材料旨在输送蛋白质以治疗 受伤的组织。但是，很少有生物材料可以独立控制多种蛋白质的递送 单一材料，限制了其效用以传递与自然伤口愈合有关的众多蛋白质 级联。控制蛋白质递送的一种策略是使用基于亲和力的生物材料，这些生物材料采用非 - 蛋白质与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 通过确定蛋白质复杂组合的定时和局部表现如何增强组织修复 影响再生过程。我们的目标是开发新的生物材料工具，以了解蛋白质如何 材料亲和力相互作用会影响蛋白质释放和活性，调节复杂的愈合反应，并 询问蛋白质表现在组织修复中的作用。我们将解决两个重要的知识差距 阻碍了有效的生物材料的蛋白质递送的开发：1）蛋白质材料亲和力如何 相互作用会影响蛋白质释放和活性？ 2）如何基于生物材料对蛋白质表现的控制 影响组织修复？我们的创新方法涉及工程新的蛋白质材料亲和力相互作用 定向进化和理性蛋白质设计。酵母表面展示将用于发展小蛋白质结构域 （即，具有高特异性和广泛亲和力与感兴趣的蛋白质结合的affibodies）。计算 建模将用于设计与蛋白质不同区域相互作用以抑制或维持 蛋白质受体结合。由此产生的膨胀阶层将使我们能够确定蛋白质如何 物质亲和力相互作用会影响蛋白质释放和不同时间尺度的活性。会产生 结合到生物材料上以调整蛋白质释放和细胞反应。使用我们的基于亲和力的图书馆 生物材料，我们将系统地研究多种蛋白质的时间表现如何影响速率 和组织修复的质量。我们将植入生物材料，以将愈合反应的关键方面恢复1） 使用中等亲和力的亲和力提供持续递送外源蛋白到损伤部位的递送和2） 使用高亲和力的亲和力在伤害部位隔离内源性蛋白质并增强，维持或 抑制蛋白活性。虽然我们的方法是灵活的和组织不可或缺的，但我们将首先在骨损伤模型中对其进行测试，但是 这是我实验室中专业知识的中心领域。通过复制伤口的复杂蛋白质表现 治愈级联 新的高度模块化生物材料，可以量身定制以编排细胞反应以治疗多重 伤害类型。我们的方法将通过创造变革性对社会产生直接的好处 新的再生医学策略有可能显着改善组织修复。