Next Generation Sequencing Facilitates Quantitative Analysis of Wild Type and Ptprz1-/- lung microvascular endothelial cell (LMVEC) Transcriptomes

Purpose: To compare NGS-derived transcriptome profiling (RNA-seq) between Wild Type and Ptprz1-/- LMVEC and identify transcripts that may be linked to the phenotypic differences observed. Methods: Total RNA from freshly cultured LMVEC was extracted using the NucleoSpin RNA Plus kit from Macherey-Nagel and the concentration and purity was assessed by Nanodrop and gel agarose 1% in TBE 0.5x. RNA from 3 independent fresh cell isolations from the Ptprz1+/+ and the Ptprz1-/- LMVEC was extracted and sent for RNAseq analysis. Novogene Co., Ltd performed the mRNA isolation, cDNA library synthesis and sequencing, and the bioinformatics analysis. Results: Pairwise comparison of gene expression of Ptprz1-/- RNA (3 samples) against Ptprz1+/+ RNA (3 samples) identified several transcripts as differentially expressed. Twenty-six genes were significantly changed (adj. p value <0.05), with 9 being upregulated and 17 downregulated. These genes are linked to angiogenesis and heart development and function. Conclusions: Our study represents the first transcriptomic analysis of Ptprz1-/- endothelial cells compared to the corresponding wild-type cells, generated by RNA-seq technology. Overall design: mRNA profiles of LMVEC isolated from wild type (WT) and Ptprz1-/- (KO) mice