The Role of D4Z4-Encoded Proteins in the Osteogenic Differentiation of Mesenchymal Stromal Cells Isolated from Bone Marrow
| Autor: | Marcel Geens, Laurence Lagneaux, Karlien Pieters, Laurence de la Kethulle de Ryhove, Céline Vanderplanck, Stephen B Wilton, Alexandra Belayew, Frédérique Coppée, Karen Sermon, Charlotte Nachtegael, Eugénie Ansseau |
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| Přispěvatelé: | Reproduction and Genetics, Basic (bio-) Medical Sciences |
| Rok vydání: | 2015 |
| Předmět: |
musculoskeletal diseases
congenital hereditary and neonatal diseases and abnormalities Polyadenylation Biology Germline Cell Line DUX4 Osteogenesis medicine Gene silencing Facioscapulohumeral muscular dystrophy Humans RNA Messenger Induced pluripotent stem cell Cells Cultured Homeodomain Proteins Osteoblasts Cell Differentiation Mesenchymal Stem Cells Cell Biology Hematology medicine.disease Alkaline Phosphatase Embryonic stem cell Molecular biology Homeobox Calcium Developmental Biology |
| Zdroj: | Stem cells and development. 24(22) |
| ISSN: | 1557-8534 |
| Popis: | Facioscapulohumeral muscular dystrophy (FSHD) is associated with an activation of the double homeobox 4 (DUX4) gene, which we previously identified within the D4Z4 repeated elements in the 4q35 subtelomeric region. The pathological DUX4 mRNA is derived from the most distal D4Z4 unit and extends unexpectedly within the flanking pLAM region, which provides an intron and polyadenylation signal. The conditions that are required to develop FSHD are a permissive allele providing the polyadenylation signal and hypomethylation of the D4Z4 repeat array compared with the healthy muscle. The DUX4 protein is a 52-kDa transcription factor that initiates a large gene deregulation cascade leading to muscle atrophy, inflammation, differentiation defects, and oxidative stress, which are the key features of FSHD. DUX4 is a retrogene that is normally expressed in germline cells and is submitted to repeat-induced silencing in adult tissues. Since DUX4 mRNAs have been detected in human embryonic and induced pluripotent stem cells, we investigated whether they could also be expressed in human mesenchymal stromal cells (hMSCs). We found that DUX4 mRNAs were induced during the differentiation of hMSCs into osteoblasts and that this process involved DUX4 and new longer protein forms (58 and 70 kDa). A DUX4 mRNA with a more distant 5' start site was characterized that presented a 60-codon reading frame extension and encoded the 58-kDa protein. Transfections of hMSCs with an antisense oligonucleotide targeting DUX4 mRNAs decreased both the 52- and 58-kDa protein levels and confirmed their identity. Gain- and loss-of-function experiments in hMSCs suggested these DUX4 proteins had opposite roles in osteogenic differentiation as evidenced by the alkaline phosphatase activity and calcium deposition. Differentiation was delayed by the 58-kDa DUX4 expression and it was increased by 52-kDa DUX4. These data indicate a role for DUX4 protein forms in the osteogenic differentiation of hMSCs. |
| Databáze: | OpenAIRE |
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