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

合作研究：GOALI：动态调节 CHO 代谢以优化生物制造产量和质量

• 批准号: 2035085
• 负责人: Jamey Young
• 金额: $ 32.6万
• 依托单位: Vanderbilt University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2035085&HistoricalAwards=false
• 关键词: Collaborative; Research; GOALI; Dynamic; regulation

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.

中国仓鼠卵巢（CHO）细胞产生所有蛋白质理论的60-70％。这些是制造所有药物中最昂贵的。确定改善单克隆抗体（MAB）生产的策略是该项目的主要目标。 CHO细胞的代谢途径将经过设计以实现这一目标。使这些疗法更加负担得起，并为公众使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Janssen研发设施的实习。这样的经验将使他们为生物技术行业或政府或学术实验室的职业做好准备。还将招募本科生和高中生，从事CHO Cell Engineering的合作。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一个关键的生物制造挑战。总体目的是设计动态调节其中心代谢的CHO细胞，以促进重组MAB的最大产量和质量。通过控制具有诱导基因开关的代谢基因的表达，将对生产阶段的工业CHO系进行设计，以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提高能效。最后，将评估IgG聚糖剖面，以确定操纵CHO糖和氨基酸代谢如何影响产品糖基化。代谢通量分析将应用于严格评估遗传改变对宿主代谢的影响，尤其是在线粒体和供应核骨糖糖前体以进行mAb糖基化的途径。该奖项反映了NSF的法定使命，并通过使用基金会的知识分子优点和更广泛的审查标准评估来诚实地获得支持。

项目成果

Attenuation of glutamine synthetase selection marker improves product titer and reduces glutamine overflow in Chinese hamster ovary cells

谷氨酰胺合成酶选择标记的减弱可提高中国仓鼠卵巢细胞中的产物滴度并减少谷氨酰胺溢出

• DOI: 10.1002/bit.28084
• 发表时间: 2022
• 期刊: Biotechnology and Bioengineering
• 影响因子: 3.8
• 作者: Sacco, Sarah A.;Tuckowski, Angela M.;Trenary, Irina;Kraft, Lauren;Betenbaugh, Michael J.;Young, Jamey D.;Smith, Kevin D.
• 通讯作者: Smith, Kevin D.

13C metabolic flux analysis in cell line and bioprocess development

• DOI: 10.1016/j.coche.2021.100718
• 发表时间: 2021-08-17
• 期刊: CURRENT OPINION IN CHEMICAL ENGINEERING
• 影响因子: 6.6
• 作者: Sacco, Sarah A.;Young, Jamey D.
• 通讯作者: Young, Jamey D.