Collaborative Research: GOALI: Dynamic regulation of CHO metabolism to optimize biomanufacturing yields and quality

中国仓鼠卵巢（CHO）细胞产生60-70%的蛋白质治疗药物。这是所有药品中制造成本最高的。确定提高单克隆抗体（mAb）生产的策略是本项目的主要目标。CHO细胞的代谢途径将被改造以实现这一目标。使公众更能负担得起并获得这些疗法将改善数百万患者的健康和生活质量。该项目还将为学生提供与行业科学家合作研究的机会，包括在杨森研发中心实习。这样的经历将为他们在生物技术行业或政府或学术实验室的职业生涯做好准备。本科生和高中生也将被招募到CHO细胞工程的协同工作。在哺乳动物细胞培养中，动态平衡高效生长和产物生物合成的代谢需求是生物制造的一个关键挑战。总体目标是设计动态调节其中心代谢的CHO细胞，以提高重组单克隆抗体的最大产量和质量。工业CHO系将通过诱导基因开关控制代谢基因的表达来促进生产阶段的线粒体呼吸。从而优化CHO宿主细胞的氨基酸代谢，减少谷氨酰胺溢出，提高能量效率。最后，将评估IgG聚糖谱，以确定操纵CHO糖和氨基酸代谢如何影响产物糖基化。代谢通量分析将用于严格评估遗传改变对宿主代谢的影响，特别是在线粒体和为单抗糖基化提供核苷酸-糖前体的途径中。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。

Chinese hamster ovary (CHO) cells produce 60–70% of all protein therapeutics. These are among the most expensive of all drugs to manufacture. Identifying strategies for improving monoclonal antibody (mAb) production is the primary objective of this project. The metabolic pathways of CHO cells will be engineered to make this happen. Making these therapies more affordable and available to the public will improve the health and quality of life of millions of patients. This project will also provide the opportunity for students to engage in collaborative research with industry scientists, including internships at a Janssen R&D facility. Such an experience will prepare them for a career in the biotechnology industry or in a government or academic lab. Undergraduate and high-school students will also be recruited to work collaboratively on CHO cell engineering. Dynamically balancing the metabolic requirements for efficient growth and product biosynthesis in mammalian cell cultures is a key biomanufacturing challenge. The overall objective is to engineer CHO cells that dynamically regulate their central metabolism to promote maximum yield and quality of recombinant mAbs. An industrial CHO line will be engineered to promote mitochondrial respiration during production phase by controlling expression of metabolic genes with inducible gene switches. Then, the amino acid metabolism of CHO host cells will be optimized to reduce glutamine overflow and increase energetic efficiency. Finally, IgG glycan profiles will be assessed to determine how manipulating CHO sugar and amino acid metabolism impacts product glycosylation. Metabolic flux analysis will be applied to rigorously evaluate the influence of genetic alterations on host metabolism, particularly within mitochondria and pathways that supply nucleotide-sugar precursors for mAb glycosylation.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.