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

1218345 Harcum中国仓鼠卵巢细胞（CHO细胞）用于生产当今市场上的大多数治疗蛋白质。特别是CHO细胞能够表达重组糖蛋白，包括单克隆抗体，这些糖蛋白需要复杂的分离后修饰。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治疗剂的消费者成本时计算（每毫克10至1000美元，典型剂量为10毫克）。糖蛋白的一个主要生产挑战是蛋白质聚集。由于糖蛋白分子的大小、不完全糖基化、高合成速率和游离巯基，糖蛋白易于聚集。幸运的是，生物工艺工程师已经发现了可减少CHO细胞培养物中蛋白质聚集的解决方案，例如培养基添加剂，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，以便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本项目转录组测序组成部分开发的转录组图谱进行研究。CHO细胞转录组将公开提供并表征，以促进CHO细胞的全基因组研究。转录组分析将被用于识别影响和响应蛋白质聚集的基因。该项目将为增加公众对治疗蛋白质和糖蛋白的可用性提供强有力的概念路线。向公众提供这一额外的高质量CHO细胞转录组数据将允许一个相称的社区知识库，这将使所有学者能够有助于我们理解生物反应器工艺对该宿主细胞系的影响。更广泛的影响还包括建立和维持受过教育、有动力的学生，特别是那些来自代表性不足的少数群体的学生，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.