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

中国仓鼠卵巢(CHO)细胞产生了所有蛋白质疗法的60%-70%。这些药物是所有药物中生产成本最高的。确定提高单抗产量的策略是该项目的主要目标。将对CHO细胞的代谢途径进行改造，以实现这一点。让公众更容易负担得起和获得这些疗法，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进行合作研究的机会，包括在扬森研发机构实习。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参与CHO细胞工程的合作工作。动态平衡哺乳动物细胞培养中有效生长和产物生物合成的代谢需求是生物制造的关键挑战。总体目标是设计CHO细胞，动态调节它们的中心代谢，以促进重组mAbs的最大产量和质量。一个工业化的CHO品系将被设计出来，通过可诱导的基因开关控制代谢基因的表达，促进生产阶段的线粒体呼吸。然后，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，将对免疫球蛋白多糖图谱进行评估，以确定操纵CHO糖和氨基酸代谢如何影响产品糖基化。代谢通量分析将用于严格评估遗传变化对宿主新陈代谢的影响，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。这一奖项反映了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。

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.