Osteopontin deficiency leads to the resolution of prostatic fibrosis and inflammation

The goals of this study is to determine transcriptomic changes in the mouse ventral prostate associated with E. coli-induced chronic inflammation and fibrosis and the alteration of this gene expression signature in osteopontin deficient mice. C57BL/J or osteopontin knockout (OPN-KO) male mice were transurethrally instilled with uropathogenic E. coli (UTI89), 2 times, 3 days apart. Control mice were instilled with saline. Ventral prostate lobes were collected 8 weeks after the first instillation. Ventral lobes were thawed and the RNA was stabilized in RNAlater (Thermo Fisher Scientific) for 1 hour. Total RNA was isolated from ventral lobes using the Qiagen RNeasy Micro kit with on-column DNAse digestion (Germantown, MD). Library preparation, quality check, RNA sequencing and bioinformatics analysis were performed at the UW-Madison Biotechnology Center. RNA quantity was measured with NanoDrop and assayed on an Agilent 2100 Bioanalyzer. The cDNA library was prepared by the TruSeq Stranded Total RNA (Gold) Library Prep with after polyA enrichment and rRNA reduction. Libraries were quantified using Qubit DNA HS kit and assayed on Agilent Tapestation DNA 1000 system. Libraries were sequenced on a NovaSeq 6000 platform.