title: Second generation sequencing of microRNA in Human Bone Cells treated with Parathyroid Hormone or Dexamethasone.
abstract: We investigated the impact of treatment with parathyroid hormone (PTH) and dexamethasone (DEX) for 2 and 24h by RNA sequencing of miRNAs in primary human bone (HOB) cells. A total of 207 million reads were obtained, and normalized absolute expression retrieved for 373 most abundant miRNAs. In naïve control cells, 7 miRNAs were differentially expressed (FDR<0.05) between the two time points. Ten miRNAs exhibited differential expression (FDR <0.05) across two time points and treatments after adjusting for expression in controls and were selected for downstream analyses. Results show significant effects on miRNA expression when comparing PTH with DEX at 2h with even more pronounced effects at 24h. Interestingly, several miRNAs exhibiting differences in expression are predicted to target genes involved in bone metabolism e.g. miR-30c2, miR-203 and miR-205 targeting RUNX2, and miR-320 targeting β-catenin (CTNNB1) mRNA expression. CTNNB1and RUNX2 levels were decreased after DEX treatment and increased after PTH treatment. Our analysis also identified 2 putative novel miRNAs in PTH and DEX treated cells at 24h. RNA sequencing showed that PTH and DEX treatment affect miRNA expression in HOB cells and that regulated miRNAs in turn are correlated with expression levels of key genes involved in bone metabolism.
authors.affiliation: Department of Medical Sciences, Uppsala University, Uppsala, 75185, Sweden; Science for Life Laboratory, Department of Medical Sciences, Uppsala University Hospital, SE-75185, Uppsala, Sweden. Electronic address: navya.laxman@medsci.uu.se.
authors.forename: Navya
authors.initials: N
authors.lastname: Laxman
authors.affiliation: Department of Medical Biochemistry and Microbiology, Uppsala University, Uppsala, 75185, Sweden.
authors.forename: Carl-Johan
authors.initials: CJ
authors.lastname: Rubin
authors.affiliation: Department of Surgical Sciences, Uppsala University, Uppsala, 75185, Sweden.
authors.forename: Hans
authors.initials: H
authors.lastname: Mallmin
authors.affiliation: Department of Surgical Sciences, Uppsala University, Uppsala, 75185, Sweden.
authors.forename: Olle
authors.initials: O
authors.lastname: Nilsson
authors.affiliation: Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, 75185, Sweden.
authors.forename: Christian
authors.initials: C
authors.lastname: Tellgren-Roth
authors.affiliation: Department of Medical Sciences, Uppsala University, Uppsala, 75185, Sweden; Science for Life Laboratory, Department of Medical Sciences, Uppsala University Hospital, SE-75185, Uppsala, Sweden.
authors.forename: Andreas
authors.initials: A
authors.lastname: Kindmark
mesh_terms: D001483:Base Sequence; D017124:Conserved Sequence / Q000235:genetics; D003907:Dexamethasone / Q000494:pharmacology*; D019143:Evolution, Molecular; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D035683:MicroRNAs / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010006:Osteoblasts / Q000187:drug effects / Q000378:metabolism; D010011:Osteocytes / Q000187:drug effects / Q000378:metabolism*; D010281:Parathyroid Hormone / Q000494:pharmacology*; D015203:Reproducibility of Results
chemical_list: D035683:MicroRNAs; D010281:Parathyroid Hormone; D003907:Dexamethasone
keywords: RNA sequencing; differential expression; miRNA; osteoblasts