Dataset Information


RNA-seq of murine mesenchymal stem cells during differentiation to osteoblasts, treated with tofacitinib or baricitinib against vehicle control

ABSTRACT: Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease, characterized by synovial hyperplasia and progressive joint destruction. For treating inflammatory diseases, such as RA, the Janus kinase (JAK)-mediated signaling pathway has emerged as a therapeutic target. Therefore, JAK inhibitors, such as tofacitinib and baricitinib, are approved for the treatment of RA. While the immunomodulatory characteristics of JAK inhibition (JAKi) are well defined, the current knowledge of how JAKi affects bone homeostasis is limited. Addressing this question becomes increasingly relevant, as current therapeutic options can slow down RA progression, but joint damage and especially perarticular bone erosion remain. Therefore, the impact of JAKi on bone homeostasis requires further elucidation. For in vitro analysis, the impact of JAK inhibitors tofacitinib and baricitinib on bone-forming osteoblasts was analyzed with respect to their mineralization capability. JAKi, by both tofacitinib and baricitinib, increased mineralization function in mesenchymal stem cells (MSCs)-derived osteoblasts. Having observed increased mineralization capability in mesenchymal stem cells (MSCs)-derived osteoblasts, RNA-seq was performed to shed light on potential signaling pathways and mediators causing this effect. For that, MSCs were plated at 6,000 cells per well in a 24-well-plate in alpha-MEM, supplemented with 10% heat-inactivated FCS and 1% P/S. After one day medium was changed to alpha-MEM, 10% heat-inactivated FCS, 1% P/S and 568 mikromolar L-Ascorbic acid 2-phosphate 351 sesquimagnesium salt hydrate, containing either tofacitinib (500 nM), baricitinib (300 nM) or DMSO as vehicle control. After osteogenic induction, RNA was isolated via phenol-chloroform extraction. At least 3 mikrogram high quality RNA were used for RNA-seq analysis with the Illumina TrueSeq Stranded mRNA Kit and sequenced on an Illumina HiSeq 2500 platform.

INSTRUMENT(S): Illumina HiSeq 2500

ORGANISM(S): Mus musculus  

SUBMITTER: Silke Frey   Susanne Adam  

PROVIDER: E-MTAB-8536 | ArrayExpress | 2020-02-12



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