RNA sequencing of mouse carotid arteries after sacubitril/valsartan treatment

Purpose: To get insight into mouse strain-dependent differences in vascular remodeling, we analyzed gene expression profiles of the ligated carotid arteries after interventions in C57BL/6J and 129X1/SvJ mice Methods: mRNA isolations from snap-frozen single carotids (n=3 per group) were conducted. Tissue homogenization by Next Advance Bullet Blender with zirconium oxide beads and Trizol. Total RNA isolation using Trizol and Qiagen RNeasy Micro kit. Results: Multiple quality controls showed a reliable RNAseq of mRNAs from the ligated carotid arteries. Specifically, there was less than 2.6% drop in reads that ranged between 26.5 to 37.7 million reads. We found a good alignment and mapping to a mouse genome (82.4 - 91.0%) in this RNAseq study. A differential gene expression analysis was done by DESeq2-1.16.1 within R-3.4.1 for data normalization and differential expression analysis with an adjusted p-value threshold of 0.05 on each set of raw expression measures. Conclusions: Our study shows significant differences in gene expression signatures in carotid arteries from two inbred mouse strains after sacubitril/valsartan treatment. We conclude that genetic background determines efficacy of the sacubitril/valsartan in mice. Overall design: Mouse carotid artery mRNA profiles of C57BL/6J and 129X1/SvJ mice after two treatment regimen of vehicle, valsartan or sacubitril/valsartan were generated by deep sequencing, in triplicate, using Illumina HiSeq2500v4 high-throughput DNA sequencer.