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

蛋白质被用来治疗各种疾病。其中包括几种癌症，肾衰竭和类风湿性关节炎，仅举几例。治疗性蛋白质的产量在过去十年中大幅增加，随着针对更多疾病的治疗方法的开发，预计未来十年还将继续增加。这些蛋白质中的大多数是从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来自大休斯顿地区的高中教师和学生。这将增强和多样化STEM准备工作的管道。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率和细胞生长。重组蛋白的过度表达会导致错误折叠蛋白的积累和蛋白毒性应激。反过来，这些又会导致全球反应的激活，最终可能会抑制蛋白质合成，降低细胞活力。这种反应在刺激蛋白质降解时达到顶峰。细胞的命运取决于参与反应的信号通路的相对激活速度。该项目团队将使用最近开发的一项内部技术，以高灵敏度和动态分辨率监测基因表达。蛋白质质量控制系统的标志物在具有代表性的重组蛋白过表达时的表达情况将被概述。结果将被用于制定策略，以重塑应激反应途径，加强质量控制，并延缓高水平重组蛋白表达细胞的细胞死亡。由于应激反应在维持细胞动态平衡方面的基础作用，这项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NSF的法定使命，并通过使用基金会的智力优势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.