An Integrated Computational and Experimental Platform for CHO-based Protein Produ

描述（由申请人提供）：摘要基于蛋白质的治疗产品对医疗保健做出了巨大贡献，并在总药物中占很大且不断增长的比例。目前，大多数生物制药产品都是使用哺乳动物细胞系生产的，特别是中国仓鼠卵巢（CHO）细胞系，其产量占所有美国许可治疗蛋白的70%以上。目前，用于蛋白质生产的大多数细胞和工艺开发策略是使用试错法进行的，其中通过实验费力地改进工艺输出。这些经验优化技术被广泛使用，因为在大多数情况下，对影响宿主细胞系中生长和蛋白质产生的潜在生物因素知之甚少。对细胞系生理学和代谢的基本了解可以大大加速和改善产品开发和制造过程。为了满足开发CHO细胞模型的商业需求，并基于我们在微生物和哺乳动物细胞中代谢建模的经验，我们启动了一项计划，以开发一个综合的计算和实验平台，用于合理设计和优化CHO细胞系中的蛋白质生产。该集成平台将我们先进的建模技术与已建立的实验技术相结合，以加快时间表并改善CHO表达系统中治疗性蛋白质生产的过程。为了证明在本研究中利用综合计算/实验方法的优势，我们打算使用代谢网络重建来表征丁酸钠补充剂（通常用于增强蛋白质表达）对CHO细胞代谢的影响，并确定替代策略，这些策略导致比生产率的类似改善，而无需防止细胞凋亡（丁酸盐治疗的不良副作用）。成功完成本提案中规定的具体目标将明确证明开发CHO细胞系中重组蛋白生产的合理方法的科学和技术可行性，该方法可用于：（a）对细胞系对环境和遗传变化的反应产生基本了解，以及（B）开发新的代谢干预措施以改善蛋白生产。公共卫生相关性：基于蛋白质的治疗产品对医疗保健做出了巨大贡献，并在整个制药市场中占很大且不断增长的比例。这些FDA批准的产品中的大多数目前使用哺乳动物细胞培养系统生产。然而，大多数细胞和过程开发策略是使用试错法进行的，使得生物制药产品开发成为昂贵且漫长的过程。优化整个过程开发的需要要求使用合理的方法，将建模技术与已建立的实验技术相结合，从根本上改变宿主细胞系的开发和治疗性蛋白质的生产方式。降低治疗性蛋白质开发和制造的成本将确保下一代药物的数量足以满足患者的需求，价格也足够低，患者可以负担得起。

DESCRIPTION (provided by applicant): Abstract Protein-based therapeutic products have contributed immensely to healthcare and constitute a large and growing percentage of the total pharmaceutical drugs. The majority of the biopharmaceutical products are currently manufactured using mammalian cell lines, in particular the Chinese hamster ovary (CHO) cell line which accounts for the production of more than 70% of all US licensed therapeutic proteins. Currently, most cell and process development strategies for protein production are performed using a trial and error approach where process outputs are improved laboriously by experimentation. These empirical optimization techniques are widely used because in most cases little is known about the underlying biological factors that impact growth and protein production in the host cell lines. A fundamental understanding of cell line physiology and metabolism can greatly accelerate and improve the process of product development and manufacturing. In response to commercial demands for developing a CHO cell model and building upon our experience with metabolic modeling in microbes and mammalian cells, we have initiated a program to develop an integrated computational and experimental platform for rational design and optimization of protein production in CHO cell lines. This integrated platform combines our advanced modeling technologies with established experimental techniques to accelerate the timelines and to improve the process of therapeutic protein production in the CHO expression system. To demonstrate the advantage of utilizing an integrated computational/experimental approach in this study, we intend to characterize the effect of sodium butyrate supplementation, commonly used to enhance protein expression, on CHO cell metabolism using a metabolic network reconstruction and identify alternative strategies that result in similar improvements in specific productivity without the need to prevent apoptosis (an undesirable side effect of butyrate treatment). Successful completion of the specific aims set forth in this proposal will provide a clear demonstration of the scientific and technical feasibility of developing a rational approach for recombinant protein production in CHO cell lines that can be used to: (a) generate fundamental understanding for cell line response to environmental and genetic changes, and (b) develop novel metabolic interventions for improved protein production. PUBLIC HEALTH RELEVANCE: Protein-based therapeutic products have contributed immensely to health care and constitute a large and growing percentage of t he total pharmaceutical market. The majority of these FDA approved products are currently manufactured using mammalian cell culture systems. However, most cell and process development strategies are performed using a trial and error approach, making biopharmaceutical product development an expensive and lengthy process. The need to optimize the overall process development calls for the use of a rational approach that combines modeling technologies with established experimental techniques to fundamentally change the way host cell lines are developed and therapeutic proteins are produced. Reducing the cost of therapeutic protein development and manufacturing would ensure that the next generation of medicines can be created in amounts large enough to meet patients' needs and at a price low enough that patients can afford them.