CAREER: Engineering Biomolecules and Cells for Oral Protein Delivery

生物技术的进步已经使非常重要的治疗性蛋白质的重组形式成为可能。口服递送这种蛋白质的想法是非常有吸引力的，因为这种给药模式是非侵入性的，具有更高的患者依从性，并且在某些情况下，可以更准确地模拟天然生理学。然而，由于在胃肠道中遇到的主要障碍-特别是苛刻的pH、快速的蛋白水解降解和低肠渗透性-蛋白质通常通过口服递送具有非常低的生物利用度，因此常规地通过侵入性方法如静脉内或皮下注射施用。在这里，将采用生物分子和细胞工程相结合的方法来克服这些瓶颈，提高口服蛋白质的输送。这一战略将需要开发一般的体外定向进化方法，以提高蛋白质在胃肠道中的稳定性，以及可调的合成生物学方法，用于工程食品级微生物口服递送这些蛋白质。这里的具体应用是创造新的蛋白质和细胞，可用于口服调节糖尿病患者的葡萄糖水平。这些相同的蛋白质工程和合成生物学方法也将构成广泛教育和培训平台的基础。在宾夕法尼亚大学当地，新的教育模块将被纳入现有的本科和研究生课程，第一个国际遗传工程机器（iGEM）团队将被创建，并将建立一个新的服务学习课程，向费城市中心的高中学生教授分子生物技术。在全国范围内，高中教师、本科生和高中生将从代表性不足的少数民族人口中招募，以开展暑期研究项目。在国际上，印度孟买的贫困街头儿童将被介绍给蛋白质工程和合成生物学的原则，使用的例子是特别相关的人口。

Advances in biotechnology have made available recombinant forms of critically important therapeutic proteins. The idea of orally delivering such proteins is extremely attractive because this mode of administration is noninvasive, has higher patient compliance, and, in some cases, can more accurately mimic native physiology. However, due to major barriers encountered in the gastrointestinal tract - particularly harsh pH, rapid proteolytic degradation, and low intestinal permeability - proteins typically have very low bioavailability via oral delivery and are therefore routinely administered through an invasive method such as intravenous or subcutaneous injection. Here, a combined biomolecular and cellular engineering approach will be employed to overcome these bottlenecks and enhance oral protein delivery. This strategy will entail the development of general in vitro directed evolution approaches to enhance protein stability in the gastrointestinal tract as well as tunable synthetic biology approaches for engineering food-grade microorganisms to deliver these proteins orally. The specific application here is to create novel proteins and cells that can be used to orally regulate glucose levels in diabetic patients. These same protein engineering and synthetic biology approaches will also form the basis for a broadly reaching educational and training platform. Locally at the University of Pennsylvania, new educational modules will be incorporated into existing undergraduate and graduate courses, the first International Genetically Engineered Machine (iGEM) team will be created, and a new service learning course will be established to teach molecular biotechnology to inner-city high school students in Philadelphia. Nationally, high school teachers, undergraduates, and high school students will be recruited from underrepresented minority populations to carry out summer research projects. And internationally, impoverished street children in Mumbai, India will be introduced to principles of protein engineering and synthetic biology using examples that are of particular relevance to that population.