Next-generation, feedback-responsive cell factories for recombinant protein manufacturing

蛋白质用于治疗各种各样的疾病。这些疾病包括几种癌症、肾衰竭和类风湿性关节炎等等。治疗性蛋白质的生产在过去十年中急剧增加，并且随着针对更多疾病的治疗方法的开发，预计在下一个十年中将继续增加。这些蛋白质大多是由哺乳动物细胞产生的，哺乳动物细胞是最有能力产生具有人类活性的蛋白质的细胞。然而，它们大规模制造蛋白质的能力也受到限制。该项目将评估一些旨在增加哺乳动物细胞蛋白质产量的策略。教育和培训机会将针对大休斯顿地区的高中教师和学生。这将加强和多样化stem准备劳动力的管道。许多复杂的重组蛋白是在哺乳动物细胞中制造的。产量取决于细胞生产力和细胞生长。重组蛋白的过表达导致错误折叠蛋白的积累和蛋白毒性应激。这些，反过来，导致全局反应的激活，最终可能抑制蛋白质合成和降低细胞活力。这种反应在刺激蛋白质降解时达到高潮。细胞的命运取决于参与反应的信号通路的相对激活率。项目团队将使用最近开发的内部技术，以高灵敏度和动态分辨率监测基因表达。在代表性重组蛋白过表达时，蛋白质质量控制系统的标记物的表达将被描述。研究结果将用于制定策略，以重塑应激反应途径，加强质量控制，并延缓表达高水平重组蛋白的细胞死亡。由于应激反应在维持细胞稳态中的基本作用，本研究的结果将影响各种细胞工程应用的设计。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Proteins are used to treat a wide variety of diseases. These include several cancers, kidney failure and rheumatoid arthritis, to name a few. Production of therapeutic proteins has increased dramatically over the last decade and is expected to continue to rise for the next decade as treatments are developed for a larger number of diseases. Most of these proteins are produced from mammalian cells, which are the most capable of producing proteins with activity in humans. However, they are also limited in their ability to manufacture protein at large scale. This project will evaluate a number of strategies designed to increase protein production in mammalian cells. Education and training opportunities will target high school teachers and students from the Greater Houston area. This will enhance and diversify the pipeline of a STEM-ready workforce.Many complex recombinant proteins are manufactured in mammalian cells. The yield depends both on cell productivity and cell growth. Overexpression of recombinant proteins causes the accumulation of misfolded proteins and proteotoxic stress. These, in turn, lead to activation of a global response that may ultimately inhibit protein synthesis and reduce cell viability. This response culminates in the stimulation of protein degradation. The fate of the cell depends on the relative rates of activation of the signaling pathways involved in the response. The project team will use a recently-developed in-house technology to monitor gene expression with high sensitivity and dynamic resolution. Expression of markers of the protein quality control system upon overexpression of representative recombinant proteins will be profiled. Results will be used to generate strategies to remodel the stress response pathway, enhance quality control, and delay cell death in cells expressing high levels of recombinant proteins. Because of the fundamental role of the stress response in maintaining cellular homeostasis, results from this study will impact the design of a variety of cell engineering applications.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.