Exploitation of the ribosomal protein L10 R98S mutation to enhance recombinant protein production in mammalian cells.

In 8% of all pediatric patients suffering from T-cell acute lymphoblastic leukemia (T-ALL), a hotspot arginine to serine mutation was found on the 98th amino-acid position (R98S) in the ribosomal protein L10 (RPL10). Preliminary research in the mouse lymphoid Ba/F3 cell model has shown that this alteration in RPL10 changes the metabolic properties within these cells. This includes increased production of freshly synthesized proteins and reduced proteasome activity, which results in an overall 25% increase in newly synthesized proteins. Data also suggest that the RPL10 R98S mutation may also increase translational fidelity and may enhance cell survival in nutrient poor conditions. Even though cell proliferation is decreased due to a ribosome biogenesis defect induced by the mutation, proper cell growth can be rescued by introducing a compensatory activating mutation in the NOTCH1 oncogene. These findings may render the RPL10 R98S mutation of interest in a biopharmaceutical setting as a means to enhance efficiency of recombinant protein production in mammalian cells. This project aims to test whether the beneficial effects of the RPL10 R98S mutation for protein production (found in the Ba/F3 model) are also present in well-established mammalian cell lines which are more relevant for protein production in large quantities, like Chinese Hamster Ovary (CHO) and Human Embryonic Kidney 293 (HEK293) cells. We intend to show higher yield gains and higher translational fidelity of a selection of 20 medically relevant proteins in the widely used CHO and HEK293 cells, genetically modified to carry the RPL10 R98S mutation. Protein activtity assays are then performed on recombinant proteins which are increasingly produced. Finally, the role of other known cancer-associated ribosomal protein mutations on recombinant protein production may be further explored.

Publication year:2022

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