Optimizing VLP production in gene therapy: Opportunities and challenges for in silico modeling

Abstract

Over the past decades, virus‐like particle (VLP)‐based gene therapy (GT) evolved as a promising approach to cure inherited diseases or cancer. Tremendous costs due to inefficient production processes remain one of the key challenges despite considerable efforts to improve titers. This review aims to link genome‐scale metabolic models (GSMMs) to cell lines used for VLP synthesis for the first time. We summarize recent advances and challenges of GSMMs for Chinese hamster ovary (CHO) cells and provide an overview of potential cell lines used in GT. Although GSMMs in CHO cells led to significant improvements in growth rates and recombinant protein (RP)‐production, no GSMM has been established for VLP production so far. To facilitate the generation of GSMM for these cell lines we further provide an overview of existing omics data and the highest production titers so far reported.

Enhancement of rAAV titers via inhibition of the interferon signaling cascade in transfected HEK293 suspension cultures

“…One of these strategies involves optimizing vector constructs by adjusting elements in Rep/Cap, Helper, and Gene of Interest (GOI) plasmid such as promoters or codon optimization to improve gene expression levels and viral packaging efficiency, ultimately increasing the overall titer. [4][5][6][7][8] Improving expression systems constitute another approach, which includes upgrading host cells, and optimizing culture conditions to achieve higher cell densities and increased productivity. Tuning transfection conditions by adjusting transfection reagents, plasmid ratios, or protocols can bolster the efficiency of DNA delivery into host cells, resulting in increased AAV production.…”