title: Gene expression profiling of bone marrow mesenchymal stem cells from Osteogenesis Imperfecta patients during osteoblast differentiation.
abstract: Mesenchymal stem cells (MSCs) are precursors present in adult bone marrow that are able to differentiate into osteoblasts, adipocytes and chondroblasts that have gained great importance as a source for cell therapy. Recently, a number of studies involving the analysis of gene expression of undifferentiated MSCs and of MSCs in the differentiation into multiple lineage processes were observed but there is no information concerning the gene expression of MSCs from Osteogenesis Imperfecta (OI) patients. Osteogenesis Imperfecta is characterized as a genetic disorder in which a generalized osteopenia leads to excessive bone fragility and severe bone deformities. The aim of this study was to analyze gene expression profile during osteogenic differentiation from BMMSCs (Bone Marrow Mesenchymal Stem Cells) obtained from patients with Osteogenesis Imperfecta and from control subjects. Bone marrow samples were collected from three normal subjects and five patients with OI. Mononuclear cells were isolated for obtaining mesenchymal cells that had been expanded until osteogenic differentiation was induced. RNA was harvested at seven time points during the osteogenic differentiation period (D0, D+1, D+2, D+7, D+12, D+17 and D+21). Gene expression analysis was performed by the microarray technique and identified several differentially expressed genes. Some important genes for osteoblast differentiation had lower expression in OI patients, suggesting a smaller commitment of these patient's MSCs with the osteogenic lineage. Other genes also had their differential expression confirmed by RT-qPCR. An increase in the expression of genes related to adipocytes was observed, suggesting an increase of adipogenic differentiation at the expense osteogenic differentiation.
authors.affiliation: Department of Genetics, Ribeirão Preto Medical School, Universidade de São Paulo, Ribeirão Preto, São Paulo, Brazil; Department of Biological Science, Santa Cruz State University, Ilhéus, BA, Brazil. Electronic address: cmkaneto@uesc.br.
authors.forename: Carla Martins
authors.initials: CM
authors.lastname: Kaneto
authors.affiliation: Department of Natural Science, Bahia State University, Vitória da Conquista, Bahia, Brazil.
authors.forename: Patrícia S
authors.initials: PS
authors.lastname: Pereira Lima
authors.affiliation: Regional Blood Center of Ribeirão Preto and National Institute of Science and Technology in Cell Therapy, Ribeirão Preto, Brazil.
authors.forename: Karen Lima
authors.initials: KL
authors.lastname: Prata
authors.affiliation: Department of Biological Science, Santa Cruz State University, Ilhéus, BA, Brazil.
authors.forename: Jane Lima
authors.initials: JL
authors.lastname: Dos Santos
authors.affiliation: Department of Genetics, Ribeirão Preto Medical School, Universidade de São Paulo, Ribeirão Preto, São Paulo, Brazil.
authors.forename: João Monteiro
authors.initials: JM
authors.lastname: de Pina Neto
authors.affiliation: Regional Blood Center of Ribeirão Preto and National Institute of Science and Technology in Cell Therapy, Ribeirão Preto, Brazil.
authors.forename: Rodrigo Alexandre
authors.initials: RA
authors.lastname: Panepucci
authors.affiliation: Department of Genetics, Ribeirão Preto Medical School, Universidade de São Paulo, Ribeirão Preto, São Paulo, Brazil.
authors.forename: Houtan
authors.initials: H
authors.lastname: Noushmehr
authors.affiliation: Regional Blood Center of Ribeirão Preto and National Institute of Science and Technology in Cell Therapy, Ribeirão Preto, Brazil.
authors.forename: Dimas Tadeu
authors.initials: DT
authors.lastname: Covas
authors.affiliation: Department of Clinical Medicine, Ribeirão Preto Medical School, Universidade de São Paulo, Ribeirão Preto, São Paulo, Brazil.
authors.forename: Francisco José Alburquerque
authors.initials: FJA
authors.lastname: de Paula
authors.affiliation: Department of Genetics, Ribeirão Preto Medical School, Universidade de São Paulo, Ribeirão Preto, São Paulo, Brazil; Regional Blood Center of Ribeirão Preto and National Institute of Science and Technology in Cell Therapy, Ribeirão Preto, Brazil.
authors.forename: Wilson Araújo
authors.initials: WA
authors.lastname: Silva
mesh_terms: D000293:Adolescent; D000328:Adult; D016022:Case-Control Studies; D002478:Cells, Cultured; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism*; D010013:Osteogenesis Imperfecta / Q000235:genetics* / Q000378:metabolism / Q000473:pathology
keywords: Bone marrow mesenchymal stem cell; Gene expression; Osteogenesis Imperfecta; Osteogenic differentiation