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

中国仓鼠卵巢（CHO）细胞产生所有蛋白质治疗剂的60-70%。这些药物是所有药物中制造成本最高的。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。使这些疗法更便宜，更容易为公众所用，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，包括在杨森研发机构实习。这样的经验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合作研究CHO细胞工程。动态平衡哺乳动物细胞培养物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。总体目标是工程化CHO细胞，其动态调节其中心代谢以促进重组mAb的最大产量和质量。将对工业CHO系进行工程改造，以通过用诱导型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。然后，将优化CHO宿主细胞的氨基酸代谢以减少谷氨酰胺溢出并增加能量效率。最后，将评估IgG聚糖谱，以确定操作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mAb糖基化提供核苷酸-糖前体的途径中。该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估来支持。

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.