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BACKGROUND: Cornelia de Lange syndrome is a genetic disease with multiple developmental defects, of which NIPBL is the main pathogenic gene. OBJECTIVE: To investigate the effect of NIPBL gene on the proliferation and osteogenic differentiation of bene marrow mesenchymal stem cells. METHODS: The NIPBL+/- mice were constructed by NIPBL-Loxp and Cre mice and used as experimental group, and the wild-type NIPBL+/+ mice served as control group. Mouse bone marrow mesenchymal stem cells were isolated and cultured in the two groups. Cell proliferation was detected using cell counting kit-8 assay when thecells were passed to the third generation. Osteoblastic differentiation was then compared between two groups after osteogenesis induction. RESULTS AND CONCLUSION: The proliferation capacity of bone marrow mesenchymal stem cells in the experimental group was lower than that in the control group (P < 0.05). The activity of alkaline phosphatase in the experimental group was significantly lower than that in the control group on the 7th day of osteogenic induction (P < 0.05). The expression levels of osteogenic genes and proteins (Runx2 and OCN) in the experimental group were lower than those in the control group after osteogenic induction (P < 0.05). On the 21st day of osteogenic induction, results from alizarin red staining indicated there were more red calcium nodules in the control group than the experimental group under inverted microscope. These findings suggest that NIPBL gene knockout can reduce the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells.
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ObjectiveThe occurrence of atrial septal defect in Cornelia DE Lange syndrome are rarely reported in previous studies. The objective of this study is to investigate the relationship between the formation of atrial septal defect and Foxa2 gene expression in NIPBL+/- mouse model through the analysis of the expression of Foxa2 gene in heart tissue. MethodsThe NIPBL+/- mice were used as the experimental group, and wild-type NIPBL+/+ mice were used as the normal group. There were six mice in each group. The weights of the mice at 1, 2, 3, and 4 weeks were measured and recorded. Mouse heart tissues were collected at 4 weeks, and the expression of Foxa2 gene and protein were determined by real-time fluorescence quantitative PCR (Qrt-PCR) and Western blot. Pathological sections and HE staining were carried out to observe the effect of NIPBL gene defect on the development and changes of the mouse heart, as well as the anatomical structure of the atrial septum of the heart. ResultsThe weight of mice in the experimental group was significantly lower than that of the normal group (P<0.05) at all four time points. Foxa2 gene expression and protein expression in heart tissues of the experimental group were statistically lower than those of the normal group (P<0.05). Abnormal defect was observed in the atrial septum of the heart pathological sections of the NIPBL deficient mice. ConclusionThe decreased expression of Foxa2 gene in the heart tissue of NIPBL+/- deficient mice may be a factor inducing growth retardation and weight loss in mice. The abnormal defect in the atrial septum of nipbl-deficient mice might be associated with the down-regulation of Foxa2 gene expression in cardiomyocytes.
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Objective@#To explore the genetic basis for an infant featuring developmental delay, hand deformity and hypertonia of extremities.@*Methods@#Clinical data and peripheral blood samples of the proband and her parents were collected. Following DNA extraction, potential mutations were screened on an Ion PGM platform using a gene panel. Suspected mutation was verified by PCR and Sanger sequencing.@*Results@#A novel heterozygous nonsense mutation, c. 2521C>T(p.R841X), was identified in the NIPBL gene. The mutation may cause premature termination of translation of the adhesion protein loading factor at 841st amino acids. The same mutation was not found in her parents and 931 healthy controls, and was absent from public databases including ExAC and 1000G. Bioinformatic analysis suggested the mutation to be disease causing.@*Conclusion@#The c. 2521C>T (p.R841X) mutation of the NIPBL gene probably underlies the Cornelia De Lange syndrome in the infant. Prenatal diagnosis may be provided to this family upon their subsequent pregnancy.
ABSTRACT
BACKGROUND: Cornelia de Lange syndrome (CdLS) is a multisystem developmental disorder in children. The disorder is caused mainly due to mutations in Nipped-B-like protein. The molecular data for CdLS is available from developed countries, but not available in developing countries like India. In the present study, the hotspot region of NIPBL gene was screened by Polymerase Chain Reaction which includes exon 2, 22, 42, and a biggest exon 10, in six CdLS patients and ten controls. MATERIALS AND METHODS: The method adopted in present study was amplification of the target exon by using polymerase chain reaction, qualitative confirmation of amplicons by Agarose Gel Electrophoresis and use of amplicons for Conformation Sensitive Gel Electrophoresis to find heteroduplex formation followed by sequencing. RESULTS: We report two polymorphisms in the studied region of gene NIPBL. The polymorphisms are in the region of intron 1 and in exon 10. The polymorphism C/A is present in intron 1 region and polymorphism T/G in exon 10. CONCLUSION: The intronic region polymorphism may have a role in intron splicing whereas the polymorphism in exon 10 results in amino acid change (Val to Gly). These polymorphisms are disease associated as these are found in CdLS patients only and not in controls.
Subject(s)
De Lange Syndrome/analysis , De Lange Syndrome/classification , De Lange Syndrome/epidemiology , De Lange Syndrome/genetics , Electrophoresis, Polyacrylamide Gel/methods , Exons , Humans , India , Polymorphism, Genetic , Proteins/classification , Proteins/genetics , Sequence Analysis, DNAABSTRACT
Cornelia de Lange Syndrome (CdLS) is a multiple congenital anomaly characterized by distinctive facial features, upper limb malformations, growth and cognitive retardation. The diagnosis of the syndrome is based on the distinctive clinical features. The etiology is still not clear. Mutations in the sister chromatid cohesion factor genes NIPBL, SMC1A (also called SMC1L1) and SMC3 have been suggested as probable cause of this syndrome. We experienced a case of newborn with CdLS showing bushy eyebrows and synophrys, long curly eyelashes, long philtrum, downturned angles of the mouth and thin upper lips, cleft palate, micrognathia, excessive body hair, micromelia of both hands, flexion contracture of elbows and hypertonicity. We detected a NIPBL gene mutation in a present neonate with CdLS, the first report in Korea.