title: Differentially expressed genes in autosomal dominant osteopetrosis type II osteoclasts reveal known and novel pathways for osteoclast biology.
abstract: Autosomal dominant osteopetrosis type II (ADO II) is a rare, heritable bone disorder characterized by a high bone mass and insufficient osteoclast activity. Mutations in the CLCN7 gene have been reported to cause ADO II. To gain novel insights into the pathways dysregulated in ADOII osteoclasts, we identified changes in gene expression in osteoclasts from patients with a heterozygous mutation of CLCN7. To do this, we carried out a transcriptomic study comparing gene expression in the osteoclasts of patients with ADO II and healthy donors. Our data show that, according to our selection criteria, 182 genes were differentially expressed in osteoclasts from patients and controls. From the 18 displaying the highest change in microarray, we confirmed differential expression for seven by qPCR. Although two of them have previously been found to be expressed in osteoclasts (ITGB5 and SERPINE2), the other five (CES1 (carboxyl esterase 1), UCHL1 (ubiquitin carboxy-terminal esterase L1, also known as ubiquitin thiolesterase), WARS (tryptophanyl-tRNA synthetase), GBP4 (guanylate-binding protein 4), and PRF1) are not yet known to have a role in this cell type. At the protein level, we confirmed elevated expression of ITGB5 and reduced expression of WARS, PRF1, and SERPINE2. Transfection of ClC-7 harboring the G215R mutation into osteoclasts resulted in an increased ITGB5 and reduced PRF1 expression of borderline significance. Finally, we observed that the ADO II patients presented a normal or increased serum level of bone formation markers, demonstrating a coupling between dysfunctional osteoclasts and osteoblasts. Sphingosine kinase 1 mRNA was expressed at the same level in ADO II and control osteoclasts. In conclusion, these data suggest that in addition to an acidification dysfunction caused by the CLCN7 mutation, a change in ITGB5, PRF1, WARS, and SERPINE2 expression could be part of the osteoclastic phenotype of ADO II.
authors: Institut National de la Santé et de la Recherche Médicale U606, Hôpital Lariboisière, Paris, France.
authors: Amélie E
authors: AE
authors: Coudert
authors: Regenerative Medicine Unit, Ospedale Pediatrico Bambino Gesù, Rome, Italy.
authors: Andrea
authors: A
authors: Del Fattore
authors: CEA-Institut de Génomique, Centre National de Genotypage, Evry, France.
authors: Céline
authors: C
authors: Baulard
authors: CEA-Institut de Génomique, Centre National de Genotypage, Evry, France.
authors: Robert
authors: R
authors: Olaso
authors: Institut National de la Santé et de la Recherche Médicale U606, Hôpital Lariboisière, Paris, France.
authors: Corinne
authors: C
authors: Schiltz
authors: 1] Institut National de la Santé et de la Recherche Médicale U606, Hôpital Lariboisière, Paris, France [2] Service de Biochimie, Hôpital Lariboisière, Paris, France.
authors: Corinne
authors: C
authors: Collet
authors: Department of Biotechnological and Applied Clinical Sciences, Università degli studi dell'Aquila, L'Aquila, Italy.
authors: Anna
authors: A
authors: Teti
authors: 1] Institut National de la Santé et de la Recherche Médicale U606, Hôpital Lariboisière, Paris, France [2] INSERM U606, Os et articulations, Bâtiment Viggo Petersen, Secteur Viole, Fédération de Rhumatologie, Hôpital Lariboisière, Paris, France.
authors: Marie-Christine
authors: MC
authors: de Vernejoul
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002265:Carboxylic Ester Hydrolases / Q000235:genetics; D016022:Case-Control Studies; D018118:Chloride Channels / Q000235:genetics / Q000378:metabolism; D005260:Female; D019204:GTP-Binding Proteins / Q000235:genetics; D006801:Humans; D039641:Integrin beta Chains / Q000235:genetics / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D020125:Mutation, Missense; D010010:Osteoclasts / Q000378:metabolism*; D010022:Osteopetrosis / Q000235:genetics* / Q000378:metabolism; D054353:Perforin; D010641:Phenotype; D052899:Pore Forming Cytotoxic Proteins / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D058258:Serpin E2 / Q000235:genetics / Q000378:metabolism; D059467:Transcriptome; D014369:Tryptophan-tRNA Ligase / Q000235:genetics / Q000378:metabolism; D043222:Ubiquitin Thiolesterase / Q000235:genetics; D055815:Young Adult
chemical_list: C097540:CLCN7 protein, human; D018118:Chloride Channels; D039641:Integrin beta Chains; D050505:Mutant Proteins; C510468:PRF1 protein, human; D052899:Pore Forming Cytotoxic Proteins; D012333:RNA, Messenger; C545486:SERPINE2 protein, human; D058258:Serpin E2; C482860:UCHL1 protein, human; C084344:integrin beta5; D054353:Perforin; D002265:Carboxylic Ester Hydrolases; C474609:CES1 protein, human; D043222:Ubiquitin Thiolesterase; D019204:GTP-Binding Proteins; D014369:Tryptophan-tRNA Ligase; C073175:WARS1 protein, human