James E. Summerton

Antisense Properties of Morpholino Oligomers

Abstract Morpholino oligomers are third generation antisense agents designed to be cost effective, water soluble and resistant to nuclease attack. They show high potency and sequence specificity and follow predictable targeting rules when used in antisense applications in cell culture.

Molecular Engine for Transporting Drugs Across Cell Membranes

Abstract A molecular engine has been developed from first principles to transport drugs from endosomes to the cytosol of cells. The engine is powered by the pH differential across the endosomal membrane, does not disrupt the endosomal membrane, and is disassembled into innocuous components after carrying out its transport function.

A k-mer based transcriptomics approach for antisense drug discovery targeting the Ewing’s family of tumors

Ewing’s sarcoma treatment failures are associated with high mortality indicating a need for new therapeutic approaches. We used a k-mer counting approach to identify cancer-specific mRNA transcripts in 3 Ewing’s Family Tumor (EFT) cell lines not found in the normal human transcriptome. Phosphorodiamidate morpholino oligomers targeting six EFT-specific transcripts were evaluated for cytotoxicity in TC-32 and CHLA-10 EFT lines and in HEK293 renal epithelial control cells. Average morpholino efficacy (EC50) was 0.66 ± 0.13 in TC-32, 0.25 ± 0.14 in CHLA-10 and 3.07 ± 5.02 µM in HEK293 control cells (ANOVA p < 0.01). Synergy was observed for a cocktail of 12 morpholinos at low dose (0.3 µM) in TC-32 cells, but not in CHLA-10 cells. Paired synergy was also observed in both EFT cell lines when the PHGDH pre-mRNA transcript was targeted in combination with XAGE1B or CYP4F22 transcripts. Antagonism was observed when CCND1 was targeted with XAGE1B or CYP4F22, or when IGFBP-2 was targeted with CCND1 or RBM11. This transcriptome profiling approach is highly effective for cancer drug discovery, as it identified new EWS-specific target genes (e.g. CYP4F22, RBM11 and IGBP-2), and predicted effective antisense agents (EC50 < 1 µM) that demonstrate both synergy and antagonism in combination therapy.