Impact of hnRNP1/2 Modulation (Or Downregulation) on Alternative Exon Splicing in WM266-4 Melanoma Cells

Impact of hnRNP1/2 Modulation (Or Downregulation) on Alternative Exon Splicing in WM266-4 Melanoma Cells

Alvin Sherman Library

This study aims to conduct an unbiased analysis of the modulation of heterogeneous nuclear ribonucleoproteins H1 and H2 (hnRNPH1/H2) on overall melanoma RNA expression in vitro. At present it is estimated that approximately 57,000 melanoma deaths occur worldwide each year. This prevalence, in conjunction with melanoma molecular heterogeneity and resulting difficulty in treatment, means that there is always a need for new treatment methods. A compound discovered in the lab, named 2155-14, has demonstrated selectivity in targeting melanoma cells by binding to hnRNPH1 and hnRNPH2, two closely related proteins that participate in RNA splicing. We performed Illumina RNA sequencing (RNA-seq) of total RNA extracted from WM266-4 melanoma cell line that underwent treatment with H1/H2 siRNA and healthy melanocytes that underwent the same treatment. Differential gene expression analysis using DESeq2 software has indicated upregulation of immune-related genes in melanoma cells, however, healthy melanocytes appear unaffected, which implies response specificity. An additional analysis of RNA-seq data using DEXSeq software has been done to evaluate any changes in the usage of exons following H1/H2 treatment, the results of which are elusory as to the extent of modulation and may shed light on the viability of these compounds in future melanoma treatment. Preliminary results showed that exon usage in TAF1C (ribosomal RNA regulator), E2F5 (tumor suppressor associated transcription factor), and either RNU6-672P (U6 small nuclear pseudogene) or ARAP1 (RHO-GAP domain) all showed splicing differences between WM266-4 cells and healthy melanocytes treated with the lead compounds.