Institutes of Physical Science and Information Technology, Anhui University, Hefei, P. R. China.
Beijing Institute of Biotechnology, Beijing, P. R. China.
Biotechnol J. 2021 Sep;16(9):e2000623. doi: 10.1002/biot.202000623. Epub 2021 Jun 19.
Chinese hamster ovary (CHO) cells with a high viable cell density (VCD), resilience to culture stress, and the capacity to continuously express recombinant proteins are highly desirable. Phosphatase and tension homology deleted on chromosome ten (PTEN) functions as a key negative regulator of the PI3K/Akt signaling pathway, mediating cell growth and survival. Its oncogenic mutant endows cells with an enhanced proliferation rate and resistance to death. In this study, the role of oncogenic PTEN C124S or G129E on the performance of CHO-K1 and CHO-IgG cells was investigated. Our results showed that CHO-K1 cells stably expressing PTEN C124S or G129E exhibited enhanced proliferation, reduced apoptosis rate, and increased transient expression of therapeutic antibodies compared to the control cells. Moreover, the stable overexpression of PTEN C124S or G129E endowed CHO-IgG cells with higher cell viability, VCD, and antibody titers (yield increased by approximately 0.77-fold) in the fed-batch culture process and enhanced their performance in response to the addition of sodium lactate. Moreover, the engineering of mutated PTEN in CHO-IgG cells did not alter antibody quality. Collectively, our data suggest that mutated PTEN is a potential target for improving the manufacture of therapeutic antibodies.
中国仓鼠卵巢（CHO）细胞具有较高的活细胞密度（VCD）、对培养压力的弹性以及持续表达重组蛋白的能力，这是非常理想的。磷酸酶和张力同源物缺失于染色体 10 号（PTEN）作为 PI3K/Akt 信号通路的关键负调节剂，介导细胞生长和存活。其致癌突变赋予细胞更高的增殖率和抗死亡能力。在这项研究中，研究了致癌性 PTEN C124S 或 G129E 对 CHO-K1 和 CHO-IgG 细胞性能的影响。我们的结果表明，与对照细胞相比，稳定表达 PTEN C124S 或 G129E 的 CHO-K1 细胞表现出增强的增殖、降低的凋亡率和增加的治疗性抗体的瞬时表达。此外，PTEN C124S 或 G129E 的稳定过表达使 CHO-IgG 细胞在补料分批培养过程中具有更高的细胞活力、VCD 和抗体滴度（产量增加约 0.77 倍），并增强了它们对添加乳酸钠的反应能力。此外，CHO-IgG 细胞中突变型 PTEN 的工程改造并未改变抗体质量。总之，我们的数据表明，突变型 PTEN 是提高治疗性抗体制造的潜在目标。
