Engineering Proteins for Reabsorption in the Renal Proximal Tubule

1264807Lazzara, Matthew J.肾脏对血浆中蛋白质的清除是导致蛋白质治疗药物生物利用度降低的最重要途径之一。这些蛋白质通过肾脏系统的损失最大的是那些不能被肾小球毛细血管壁的过滤装置有效保留的蛋白质，以及不能在近端小管内从原尿中有效重吸收的蛋白质。虽然一些基于蛋白质的治疗方法已经通过共价修饰或融合到大体积惰性蛋白质来减少肾脏清除率，但这种修饰可能会改变对预期目标组织的治疗有效性。研究人员建议在白蛋白中识别最小的表位，通过近端上皮中表达的同源转运受体触发这种丰富的血浆蛋白的近端小管重吸收，并利用这些信息来设计通用肽，这些肽可以融合到任何治疗性蛋白中，通过引起其完整的重吸收来减少其肾脏清除率。实验目标将利用调查者内部的工具？实验室用于蛋白质工程，分析蛋白质与细胞和受体的相互作用，并测量细胞内蛋白质运输过程。计算目标将利用质量传递和动力学建模方法来创建一个定量框架，以预测蛋白质结合和内吞作用的变化如何提高治疗性生物利用度，以及如何优化蛋白质工程。此外，该项目将通过综合教育计划产生更广泛的影响，其中研究目标背后的科学将在多种教育环境中传播给学生和教师。基于蛋白质的治疗方法由于其精致的生物学功能和特异性而变得越来越普遍。调查人员吗?这项工作将对提高治疗性蛋白质的功效具有广泛的意义。事实上，模型蛋白完整重吸收的成功工程将使这项技术能够应用于任何基于蛋白质的治疗，并为基于蛋白质的药物创造一种全新的模块化。通过提高治疗效果和患者治疗方案，以及减少与蛋白质类药物的生产和管理相关的经济负担，这有可能产生巨大的社会影响。

1264807Lazzara, Matthew J. The clearance of proteins from plasma by the kidneys constitutes one of the most significant pathways leading to reduced bioavailability of protein-based therapeutics. The loss of such proteins via the renal system is greatest for proteins which are not efficiently retained by the filtration apparatus of the glomerular capillary wall and which are not efficiently reabsorbed from the primary urine within the proximal tubule. While some protein-based therapeutics have been engineered for reduced renal clearance by covalent modification or fusion to bulky inert proteins, such modifications may alter therapeutic availability to intended target tissues. The investigators propose to identify minimal epitopes within albumin that trigger the efficient proximal tubule reabsorption of this abundant plasma protein through cognate transport receptors expressed in the proximal epithelium and to use this information to engineer generic peptides that can be fused to any therapeutic protein to reduce its renal clearance by causing its intact reabsorption. The experimental aims will leverage tools within the investigators? laboratories to engineer proteins, analyze protein interactions with cells and receptors, and measure intracellular protein trafficking processes. The computational aim will leverage mass transport and kinetic modeling approaches to create a quantitative framework for predicting how changes to protein binding and endocytosis will improve therapeutic bioavailability and how to optimally engineer proteins. In addition, this project will have broader impact through integrated educational programs in which the science behind the research objectives will be disseminated to students and teachers in multiple educational settings.Protein-based therapeutics are becoming increasingly common due to their exquisite biological functions and specificities. The investigators? work will have broad implications for enhancing therapeutic protein efficacy. Indeed, the successful engineering of the intact reabsorption of model proteins will enable the application of this technology to any protein-based therapeutic and create a fundamentally new modularity for protein-based drugs. This has potential for tremendous societal impact through enhancement of therapeutic efficacy and patient treatment regimens, as well as reduction in the financial burden associated with production and administration of protein-based drugs.