Overexpression of transcription factor BLIMP1/prdm1 leads to growth inhibition and enhanced secretory capacity in Chinese hamster ovary cells.
Overexpression of transcription factor BLIMP1/prdm1 leads to growth inhibition and enhanced secretory capacity in Chinese hamster ovary cells.
复制标题
转录因子 BLIMP1/prdm1 的过度表达会导致中国仓鼠卵巢细胞的生长抑制和分泌能力增强。
DOI：
10.1016/j.ymben.2021.07.004
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发表时间：
2021
影响因子：
8.4
通讯作者：
Torres M
中科院分区：
文献类型：
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作者：
Torres M
中国仓鼠卵巢（CHO）细胞在加工和分泌大量重组蛋白方面存在固有的局限性，特别是那些需要复杂的翻译后加工的重组蛋白。为了解决这些限制，我们设计了CHO宿主细胞（CHOK1和CHOS）来过度表达转录因子BLIMP1/prdm1（产生抗体的浆细胞高分泌表型的主要调节因子），产生新的CHO细胞系（称为CHOB）。CHOB细胞系表现出细胞密度降低，静止期延长，G1/G0期细胞周期停滞，但同时重组IgG1的产物滴度显著提高（>增加2倍），细胞特异性生产力显著提高（>增加3倍）。我们证明了CHOB细胞生产表型的改善是由细胞生理和代谢的一系列变化引起的。与对照细胞相比，CHOB细胞显示出更大的内质网大小和更高的蛋白质合成和分泌能力。此外，CHOB细胞表现出有利于能量生产的代谢特征，以支持重组蛋白生产的增加。该研究表明，基于BLIMP1表达的细胞工程方法在提高CHO细胞宿主的分泌能力方面具有很大的潜力，可用于生产重组生物制药。我们的研究结果也为细胞生长与生产力之间的关系提供了更好的理解，为在工业细胞系发育过程中改善CHO细胞系的生产表型提供了有价值的通用信息。
Chinese hamster ovary (CHO) cells present inherent limitations for processing and secretion of large amounts of recombinant proteins, especially for those requiring complex post-translational processing. To tackle these limitations, we engineered CHO host cells (CHOK1 and CHOS) to overexpress the transcription factor BLIMP1/prdm1 (a master regulator of the highly-secreting phenotype of antibody-producing plasma cells), generating novel CHO cell lines (referred to as CHOB). The CHOB cell lines exhibited decreased cell densities, prolonged stationary phase and arrested cell cycle in G1/G0 phase but simultaneously had significantly greater product titre for recombinant IgG1 (> 2-fold increase) coupled with a significantly greater cell-specific productivities (> 3-fold increase). We demonstrated that the improved productive phenotype of CHOB cells resulted from a series of changes to cell physiology and metabolism. CHOB cells showed a significantly greater ER size and increased protein synthesis and secretion capacity compared to control cells. In addition, CHOB cells presented a metabolic profile that favoured energy production to support increased recombinant protein production. This study indicated that a cell engineering approach based on BLIMP1 expression offers great potential for improving the secretory capacity of CHO cell hosts utilised for manufacture of recombinant biopharmaceuticals. Our findings also provides a greater understanding of the relationship between cell growth and productivity, valuable generic information for improving productive phenotypes for CHO cell lines during industrial cell line development.