Comprehensive analysis of transcriptomic changes in cutaneous squamous cell carcinoma

Purpose: We report the coding and non-coding transcriptomic changes in primary cutaneous squamous cell carcinoma Methods: 4 mm punch biopsies were collected from 7 unmatched healthy and 9 cSCC patients and snap frozen. The library preparation was done using Illumina TruSeq® Stranded Total RNA (with Ribo-Zero Gold) preparation kit. 100 ng of total RNA was depleted of rRNAs using ribo-zero gold magnetic bead based capture-probe system (Illumina Inc.). The remaining RNA was subsequently purified (RNAcleanXP, Beckman Coulter) and fragmented using enzymatic fragmentation. Then first strand synthesis and second strand synthesis were performed and the double stranded cDNA was purified (AMPure XP, Beckman Coulter). The cDNA was end repaired, 3' adenylated and Illumina sequencing adaptors ligated onto the fragments ends, and the library was purified (AMPure XP). The stranded libraries were amplified with PCR and purified (AMPure XP). The libraries size distribution was validated and quality inspected on a Bioanalyzer. Sequencing was performed on NextSeq 500 instrument using v2 reagent kits according to the manufacturer instructions (Illumina Inc.). Results: On average 49.8 million 100 base pair (bp) paired-end reads were obtained from each sample and genome mapping was on average 55% for all samples. Altered expression of 5,352 protein-coding genes, 908 lncRNAs and 55 circular RNAs was identified. Conclusions: Here, we report the coding and non-coding transcriptomic changes from cSCC samples, along with identification of skin specific transcripts and novel deregulated circRNAs. Overall design: Whole transcriptome profile of 9 cSCC and 7 unmatched healthy controls