Osteoblastic differentiation of bone marrow mesenchymal stromal cells in Bruck Syndrome

Autor: Francisco José Albuquerque de Paula, Carla Martins Kaneto, João Monteiro de Pina Neto, Francisco de Assis Pereira, Wilson A. Silva, Julio C. C. Lorenzi, Karen de Lima Prata, Dalila Luciola Zanette, Jane Lima dos Santos, Patricia Santos Pereira Lima, Thiago Y. Oliveira
Rok vydání: 2016
Předmět:
0301 basic medicine
Adult
Genetic Markers
Male
musculoskeletal diseases
Adolescent
macromolecular substances
030105 genetics & heredity
Biology
03 medical and health sciences
Young Adult
Bruck syndrome
N-terminal telopeptide
Bone Marrow
Osteogenesis
Osteogenic differentiation
medicine
Genetics
Humans
Genetics(clinical)
Child
Genetics (clinical)
Cells
Cultured
Arthrogryposis
Osteoblasts
Gene Expression Profiling
Mesenchymal stem cell
EXPRESSÃO GÊNICA
Cell Differentiation
Mesenchymal Stem Cells
Sequence Analysis
DNA
Osteogenesis Imperfecta
medicine.disease
Phenotype
Bone marrow mesenchymal stromal cell
Procollagen peptidase
Collagen
type I
alpha 1
030104 developmental biology
medicine.anatomical_structure
Gene Expression Regulation
Osteogenesis imperfecta
Cancer research
Female
Bone marrow
Gene expression
Research Article
Zdroj: Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual)
Universidade de São Paulo (USP)
instacron:USP
BMC Medical Genetics
Popis: Background Osteogenesis Imperfecta (OI) (OMIM %259450) is a heterogeneous group of inherited disorders characterized by increased bone fragility, with clinical severity ranging from mild to lethal. The majority of OI cases are caused by mutations in COL1A1 or COL1A2. Bruck Syndrome (BS) is a further recessively-inherited OI-like phenotype in which bone fragility is associated with the unusual finding of pterygia and contractures of the large joints. Notably, several studies have failed to show any abnormalities in the biosynthesis of collagen 1 in BS patientes. Evidence was obtained for a specific defect of the procollagen telopeptide lysine hydroxylation in BS, whereas mutations in the gene PLOD2 have been identified. Recently, several studies described FKBP10 mutations in OI-like and BS patients, suggesting that FKBP10 is a bonafide BS locus. Methods We analyzed the coding region and intron/exon boundaries of COL1A1, COL1A2, PLOD2 and FKBP10 genes by sequence analysis using an ABI PRISM 3130 automated sequencer and Big Dye Terminator Sequencing protocol. Mononuclear cells obtained from the bone marrow of BS, OI patients and healthy donors were cultured and osteogenic differentiation was induced. The gene expression of osteoblast specific markers were also evaluated during the osteoblastic differentiation of mesenchymal stem cell (MSC) by qRT-PCR using an ABI7500 Sequence Detection System. Results No mutations in COL1A1, COL1A2 or PLOD2 were found in BS patient. We found a homozygous 1-base-pair duplication (c.831dupC) that is predicted to produce a translational frameshift mutation and a premature protein truncation 17 aminoacids downstream (p.Gly278ArgfsX95). The gene expression of osteoblast specific markers BGLAP, COL1A1, MSX2, SPARC and VDR was evaluated by Real Time RT-PCR during differentiation into osteoblasts and results showed similar patterns of osteoblast markers expression in BS and healthy controls. On the other hand, when compared with OI patients, the expression pattern of these genes was found to be different. Conclusions Our work suggests that the gene expression profiles observed during mesenchymal stromal cell differentiation into osteoblast are distinct in BS patients as compared to OI patients. The present study shows for the first time that genes involved in osteogenesis are differentially expressed in BS and OI patients. Electronic supplementary material The online version of this article (doi:10.1186/s12881-016-0301-7) contains supplementary material, which is available to authorized users.
Databáze: OpenAIRE
Externí odkaz: