CAREER: Engineering Biomolecules and Cells for Oral Protein Delivery

生物技术的进步使至关重要的治疗性蛋白质的重组形式成为可能。口服这种蛋白质的想法非常有吸引力，因为这种给药模式是非侵入性的，患者依从性更高，在某些情况下，可以更准确地模拟本地生理。然而，由于胃肠道遇到的主要障碍--特别是苛刻的pH值、快速的蛋白质降解和低的肠道通透性--蛋白质通过口服给药的生物利用度通常很低，因此通常通过静脉或皮下注射等侵入性方法给药。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，并增强口服蛋白质的输送。这一战略将需要开发通用的体外定向进化方法来增强胃肠道中蛋白质的稳定性，以及可调整的合成生物学方法来设计食品级微生物来口头传递这些蛋白质。这里的具体应用是创造新的蛋白质和细胞，可以用来口服调节糖尿病患者的血糖水平。这些同样的蛋白质工程和合成生物学方法也将构成一个广泛传播的教育和培训平台的基础。在宾夕法尼亚大学，新的教育模块将被纳入现有的本科和研究生课程，将创建第一个国际基因工程机器(IGEM)团队，并将设立一门新的服务学习课程，向费城市中心的高中生教授分子生物技术。在全国范围内，将从未被充分代表的少数民族中招募高中教师、本科生和高中生开展暑期研究项目。在国际上，印度孟买的贫困街头儿童将使用与该人群特别相关的例子，向他们介绍蛋白质工程和合成生物学的原理。

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.