Characterization of anOsmrConditional Knockout Mouse Model

Abstract

ABSTRACTOncostatin M (OSM) is a member of the interleukin-6 (IL-6) family of cytokines and has been found to have distinct anti-inflammatory and pro-inflammatory properties in various cellular and disease contexts. OSM signals through two receptor complexes, one of which includes OSMRβ. To investigate OSM-OSMRβ signaling in adult hematopoiesis, we utilized the readily available conditionalOsmrfl/flmouse model B6;129-Osmrtm1.1Nat/J, which is poorly characterized in the literature. This model contains loxP sites flanking exon 2 of theOsmrgene. We crossedOsmrfl/flmice to interferon-inducibleMx1-Cre, which is robustly induced in adult hematopoietic cells. We observed complete recombination of theOsmrflallele and loss of exon 2 in hematopoietic (bone marrow) as well as non-hematopoietic (liver, lung, kidney) tissues. Using a TaqMan assay with probes downstream of exon 2,Osmrtranscript was lower in the kidney but equivalent in bone marrow, lung, and liver fromOsmrfl/flMx1-Cre versusMx1-Cre control mice, suggesting that transcript is being produced despite loss of this exon. Western blots show that liver cells fromOsmrfl/flMx1-Cre mice had complete loss of OSMR protein, while bone marrow, kidney, and lung cells had reduced OSMR protein at varying levels. RNA-seq analysis of a subpopulation of bone marrow cells (hematopoietic stem cells) finds that some OSM-stimulated genes, but not all, are suppressed inOsmrfl/flMx1-Cre cells. Together, our data suggest that the B6;129-Osmrtm1.1Nat/J model should be utilized with caution as loss ofOsmrexon 2 has variable and tissue-dependent impact on mRNA and protein expression.