EAGER: Transcriptome analysis of CHO cells to improve productivity and control protein aggregation

1218345中国仓鼠卵巢细胞(CHO细胞)被用来制造目前市场上的大部分治疗性蛋白。特别是，CHO细胞能够表达重组糖蛋白，包括需要复杂的翻译后修饰的单抗。尽管CHO细胞已成为新的“大肠杆菌”主力细胞系，但仍有许多工程挑战必须解决。这些挑战可以在评估糖蛋白疗法消费者的成本时计算出来(每毫克S 10美元到S 1,000美元，典型剂量为10‘S毫克)。糖蛋白生产的一个主要挑战是蛋白质聚集。糖蛋白由于分子大小、不完全糖基化、高合成速率和游离硫醇而易于聚集。幸运的是，生物过程工程师已经发现了解决方案，如介质添加剂，可以减少CHO细胞培养中的蛋白质聚集，但更有效的长期解决方案是稳定生产和降低成本所必需的。本研究项目的总体目标是开发和增强工具箱，从基因表达开始更好地理解蛋白质聚集。导致蛋白质聚集的因素和减少蛋白质聚集的条件将通过询问从该项目的转录组测序部分发展而来的转录组图谱来调查。CHO细胞转录组将公之于众，并将用于促进CHO细胞的全基因组研究。转录组分析将用于确定影响和响应蛋白质聚集的基因。该项目将为增加治疗蛋白和糖蛋白对公众的可用性提供强有力的概念性途径。这些额外的高质量CHO细胞转录组数据的公开将允许一个相应的社区知识库，使所有学者能够帮助我们理解生物反应器过程对宿主细胞系的影响。更广泛的影响还包括建立和维持一系列受过教育、积极进取的学生，特别是那些来自代表性不足的少数群体的学生，他们从事生物技术职业，以满足美国日益增长的需求。

1218345HarcumChinese hamster ovary cells (CHO cells) are used to manufacture a majority of the therapeuticproteins on the market today. In particular, CHO cells are able to express recombinantglycoproteins, including monoclonal antibodies, which require complex post-translationalmodifications. Even though CHO cells have become the new "Escherichia coli" workhorse cellline, there are still many engineering challenges that must be addressed. These challenges canbe calculated when assessing the cost to consumers of glycoprotein therapeutics ($10's to$1000's of dollars (US) per milligram with typical doses of 10's of milligrams).One major production challenge for glycoproteins is protein aggregation. Glycoproteins areprone to aggregation due to the size of these molecules, incomplete glycosylation, highsynthesis rates, and free thiols. Fortunately, bioprocess engineers have discovered interimsolutions, such as, media additives that can reduce protein aggregation in CHO cell cultures,but more efficient long-term solutions are necessary to stabilize production and reduce costs.The overall goal of this research project is to develop and enhance the toolbox for betterunderstanding protein aggregation beginning with gene expression. Factors that lead to proteinaggregation and conditions that reduce protein aggregation will be investigated by interrogatingthe transcriptome profiles developed from the transcriptome-sequencing component of thisproject. The CHO cell transcriptome will be made publicly available and characterized topromote genome-wide research in CHO cells. Transcriptome analysis will be used to identifygenes that affect and respond to protein aggregation.This project will provide a strong conceptual route for increasing the availability of therapeuticproteins and glycoproteins to the public. The availability of this additional high quality CHO celltranscriptome data to the public will allow for a commensurate community knowledge base thatwill enable all scholars the ability to contribute to our understanding of bioreactor process effectson this host cell line. Broader impacts also include the establishment and maintenance of apipeline of educated, motivated students, particularly those from under-represented minoritypopulations to undertake careers in biotechnology to meet this growing demand in the US.