RNA-seq study of a helicon peptide targeting ß-Catenin/TCF transcription factor [in vitro]

Genetic and epigenetic alterations in the Wnt signaling pathway leading to constitutive activation of the driver oncogene ß-catenin occur in at least 20% of all human cancers. Pharmacologic suppression of aberrant ß-catenin transcriptional activity through direct targeting of its downstream effectors has proven elusive despite decades of effort. We developed conformationally hyperstabilized a-helical peptides – which we refer to as Helicons – that directly bind ß-catenin with sub-nanomolar affinity and exhibit good cytosolic exposure. COLO320DM is a human colorectal cancer line with an activated Wnt/ ß-catenin pathway driven by an APC mutation. Here, we characterize the global effects of Helicon treatment on COLO320DM cells in vitro by whole-transcriptome RNA sequencing. As expected, Helicon 4 treatment had both time- and dose-dependent effects on the COLO320DM transcriptional profile. In particular, the Hallmark Wnt/ß-catenin gene set was significantly down-regulated as early as six-hour at 10uM. The result validates the on-target inhibition of ß-catenin signaling through disruption of its interaction with TCF/LEF transcription factors by active Helicons. COLO320DM is a human colorectal cancer line with an activated Wnt/ ß-catenin pathway driven by an APC mutation. Here, we characterized the global effects of Helicon treatment on COLO320DM cells in vitro by whole-transcriptome RNA sequencing. As expected, Helicon 4 treatment had both time- and dose-dependent effects on the COLO320DM transcriptional profile. In particular, the Hallmark Wnt/ß-catenin gene set was significantly down-regulated in as early as six -hours at 10 µuM. The result validates the on-target inhibition of ß-catenin signaling through disruption of its interaction with TCF/LEF transcription factors by active Helicons. Overall design: COLO320DM cell line was maintained in RPMI with 4%FBS. Cells were treated by Helicon 4, Helicon 4-neg or DMSO at indicated dose and timepoint, followed by RNA extraction and Sequencing