RESUMO
Expression of placental growth factor (PGF) is closely associated with placental perfusion in early pregnancy. PGF is primarily expressed in placental trophoblasts, and its expression decreases in preeclampsia, associated with placental hypoxia. The transcription factors glial cells missing 1 (GCM1) and metal-regulatory transcription factor 1 (MTF1) have been implicated in the regulation of PGF gene expression through regulatory elements upstream and downstream of the PGF transcription start site, respectively. Here, we clarified the mechanism underlying placenta-specific PGF expression. We demonstrate that GCM1 up-regulates PGF expression through three downstream GCM1-binding sites (GBSs) but not a previously reported upstream GBS. Interestingly, we also found that these downstream GBSs also harbor metal-response elements for MTF1. Surprisingly, however, we observed that MTF1 is unlikely to regulate PGF expression in the placenta because knockdown or overexpression of GCM1, but not MTF1, dramatically decreased PGF expression or reversed the suppression of PGF expression under hypoxia, respectively. We also demonstrate that another transcription factor, Distal-less homeobox 3 (DLX3), interacts with the DNA-binding domain and the first transactivation domain of GCM1 and that this interaction inhibits GCM1-mediated PGF expression. Moreover, the GCM1-DLX3 interaction interfered with CREB-binding protein-mediated GCM1 acetylation and activation. In summary, we have identified several GBSs in the PGF promoter that are highly responsive to GCM1, have demonstrated that MTF1 does not significantly regulate PGF expression in placental cells, and provide evidence that DLX3 inhibits GCM1-mediated PGF expression. Our findings revise the mechanism for GCM1- and DLX3-mediated regulation of PGF gene expression.
Assuntos
Regulação da Expressão Gênica , Proteínas de Homeodomínio/metabolismo , Proteínas Nucleares/metabolismo , Fator de Crescimento Placentário/genética , Placenta/metabolismo , Elementos de Resposta , Fatores de Transcrição/metabolismo , Trofoblastos/metabolismo , Acetilação , Sequência de Bases , Diferenciação Celular , Proteínas de Ligação a DNA , Feminino , Proteínas de Homeodomínio/genética , Humanos , Proteínas Nucleares/genética , Fator de Crescimento Placentário/metabolismo , Gravidez , Regiões Promotoras Genéticas , Ligação Proteica , Fatores de Transcrição/genéticaRESUMO
OBJECTIVE: Distal-less homeobox 3 (DLX3) is an important transcription factor involved in the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). However, the underlying mechanism is not clear. This study investigated the underlying mechanism of DLX3 in osteogenic differentiation. METHODS: DLX3 overexpression and knockdown in cells were achieved using lentiviruses. The osteogenic differentiation of BMSCs was detected using alkaline phosphatase expression, alizarin red staining, real-time quantitative polymerase chain reaction (RT-qPCR), Western blotting, and chromatin immunoprecipitation (ChIP) assays. RESULTS: DLX3 overexpression promoted the osteogenic differentiation of BMSCs, whereas DLX3 knockdown reduced the osteogenic differentiation of BMSCs. RT-qPCR and Western blotting assays showed that DLX3 modulated osteogenic differentiation via the Wnt/ß-catenin pathway. ChIP-qPCR showed that DLX3 knockdown promoted DKK4 expression by decreasing the enrichment of histone H3 lysine 27 trimethylation (H3K27me3) in the promotor region of DKK4. CONCLUSION: Our data implied that DLX3 regulated Wnt/ß-catenin pathway through histone modification of DKK4 during the osteogenic differentiation of BMSCs.
Assuntos
Histonas/metabolismo , Proteínas de Homeodomínio/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Células-Tronco Mesenquimais/citologia , Osteogênese , Fatores de Transcrição/metabolismo , Via de Sinalização Wnt , Diferenciação Celular , Células Cultivadas , Humanos , Células-Tronco Mesenquimais/metabolismo , MetilaçãoRESUMO
In a previous study, we showed that microRNA-675 (miR-675) was significantly down-regulated in patients with tricho-dento-osseous (TDO) syndrome. One of the main features of TDO syndrome is dentin hypoplasia. Thus, we hypothesize that miR-675 plays a role in dentin development. In this study, we determined the role of miR-675 in the odontogenic differentiation of human dental pulp cells (hDPCs). Stable overexpression and knockdown of miR-675 in hDPCs were performed using recombinant lentiviruses containing U6 promoter-driven miR-675 and short hairpin-miR675 expression cassettes, respectively. Alkaline phosphatase (ALP) assay, Alizarin red staining assay, quantitative polymerase chain reaction (qPCR), Western blot analysis, and immunofluorescent staining revealed the promotive effects of miR-675 on the odontogenic differentiation of hDPCs. Further, we found that miR-675 facilitates the odontogenic differentiation process of hDPCs by epigenetic regulation of distal-less homeobox (DLX3). Thus, for the first time, we determined that miR-675 regulates the odontogenic differentiation of hDPCs by inhibiting the DNA methyltransferase 3 beta (DNMT3B)-mediated methylation of DLX3. Our findings uncover an unanticipated regulatory role for miR-675 in the odontogenic differentiation of hDPCs by epigenetic changes in DLX3 and provide novel insight into dentin hypoplasia feature in TDO patients.
Assuntos
Diferenciação Celular/genética , Anormalidades Craniofaciais/genética , Hipoplasia do Esmalte Dentário/genética , Polpa Dentária/citologia , Epigênese Genética/genética , Doenças do Cabelo/genética , Proteínas de Homeodomínio/metabolismo , MicroRNAs/metabolismo , Fatores de Transcrição/metabolismo , Anormalidades Craniofaciais/metabolismo , Anormalidades Craniofaciais/patologia , DNA (Citosina-5-)-Metiltransferases/genética , DNA (Citosina-5-)-Metiltransferases/metabolismo , Hipoplasia do Esmalte Dentário/metabolismo , Hipoplasia do Esmalte Dentário/patologia , Polpa Dentária/metabolismo , Dentina/metabolismo , Dentina/patologia , Técnicas de Silenciamento de Genes , Doenças do Cabelo/metabolismo , Doenças do Cabelo/patologia , Proteínas de Homeodomínio/genética , Humanos , MicroRNAs/genética , Cultura Primária de Células , Fatores de Transcrição/genética , DNA Metiltransferase 3BRESUMO
OBJECTIVES: Variants in DLX3 cause tricho-dento-osseous syndrome (TDO, MIM #190320), a systemic condition with hair, nail and bony changes, taurodontism and amelogenesis imperfecta (AI), inherited in an autosomal dominant fashion. Different variants found within this gene are associated with different phenotypic presentations. To date, six different DLX3 variants have been reported in TDO. The aim of this paper was to explore and discuss three recently uncovered new variants in DLX3. SUBJECTS AND METHODS: Whole-exome sequencing identified a new DLX3 variant in one family, recruited as part of an ongoing study of genetic variants associated with AI. Targeted clinical exome sequencing of two further families revealed another new variant of DLX3 and complete heterozygous deletion of DLX3. For all three families, the phenotypes were shown to consist of AI and taurodontism, together with other attenuated features of TDO. RESULTS: c.574delG p.(E192Rfs*66), c.476G>T (p.R159L) and a heterozygous deletion of the entire DLX3 coding region were identified in our families. CONCLUSION: These previously unreported variants add to the growing literature surrounding AI, allowing for more accurate genetic testing and better understanding of the associated clinical consequences.
Assuntos
Amelogênese Imperfeita/genética , Anormalidades Craniofaciais/genética , Hipoplasia do Esmalte Dentário/genética , Doenças do Cabelo/genética , Proteínas de Homeodomínio/genética , Fatores de Transcrição/genética , Feminino , Humanos , Masculino , LinhagemRESUMO
Well-regulated epidermal homeostasis depends on the function of different classes of factors, such as transcription regulators and receptors. Alterations in this homeostatic balance may lead to the development of cutaneous squamous tumorigenesis. The homeobox transcription factor DLX3 is determinant for a p53-dependent regulation of epidermal differentiation and modulates skin carcinogenesis. The maintenance of skin homeostasis also involves the action of neurotrophins (NTs) and their receptors, Trk and CD271. While Trk receptor overexpression is a hallmark of cancer, there are conflicting data on CD271 expression and function in cutaneous SCC (cSCC). Previous studies have reported NT receptors expression in head and neck SSC (HNSCC). We show that CD271 is expressed at low levels in primary cSCC cells and the number of CD271+ cells correlates with cell cohesion in SCC spheroids. In normal epidermis, CD271 is expressed in proliferative progenitor cells and DLX3 in terminally differentiated keratinocytes. Brain-derived neurotrophic factor (BDNF) and neurotrophin 3 (NT3) increase DLX3 expression. In the absence of a functional BDNF receptor TrkB in keratinocytes, we hypothesize that the BDNF-dependent DLX3 response could be mediated via CD271. Altogether, our results support a putative CD271-DLX3 connection in keratinocytes, which might be crucial to preventing squamous skin cancer.
Assuntos
Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/metabolismo , Proteínas de Homeodomínio/genética , Queratinócitos/metabolismo , Proteínas do Tecido Nervoso/genética , Receptores de Fator de Crescimento Neural/genética , Fatores de Transcrição/genética , Biomarcadores , Linhagem Celular Tumoral , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Proteínas de Homeodomínio/metabolismo , Humanos , Modelos Biológicos , Proteínas do Tecido Nervoso/metabolismo , Ligação Proteica , Receptores de Fator de Crescimento Neural/metabolismo , Transdução de Sinais , Fatores de Transcrição/metabolismoRESUMO
DLX3 is essential for tooth enamel development and is so far the only transcription factor known to be mutated in a syndromic form of amelogenesis imperfecta. Through conditional deletion of Dlx3 in the dental epithelium in mouse, we have previously established the involvement of DLX3 in enamel pH regulation, as well as in controlling the expression of sets of keratins that contribute to enamel rod sheath formation. Here, we show that the decussation pattern of enamel rods was lost in conditional knockout animals, suggesting that DLX3 controls the coordinated migration of ameloblasts during enamel secretion. We further demonstrate that DLX3 regulates the expression of some components of myosin II complexes potentially involved in driving the movement of ameloblasts that leads to enamel rod decussation.
Assuntos
Ameloblastos/metabolismo , Esmalte Dentário/metabolismo , Proteínas de Homeodomínio/metabolismo , Queratinas/biossíntese , Fatores de Transcrição/metabolismo , Ameloblastos/citologia , Animais , Movimento Celular , Proteínas de Homeodomínio/genética , Concentração de Íons de Hidrogênio , Queratinas/genética , Camundongos , Camundongos Transgênicos , Fatores de Transcrição/genéticaRESUMO
Homeodomain gene Distal-less-3 (DLX3) plays an essential role in the development of bones. Mutations of DLX3 are closely associated with Tricho-Dento-Osseous (TDO) syndrome featured with increased bone formation. However, the mechanism regarding whether DLX3 regulates osteoclastogenesis remains largely unknown. In this study, we firstly examined the expression of DLX3 mounting during osteoclastic differentiation process, and then established stably expressing wild type DLX3 (WT-DLX3), a novel mutant DLX3 (Q178R) found in our laboratory recently (MT-DLX3) and Dlx3 knockdown cell lines (Dlx3-shRNA) in Raw 264.7 cells using corresponding lentiviruses. Next, we investigated the influence of DLX3 on these stable cells in the process of osteoclastogenesis. The results showed that the expression of osteoclastogenesis-related genes as well as tartrate-resistant acid phosphatase-positive multinucleated cells were lower in WT-DLX3 and MT-DLX3, but higher in Dlx3-shRNA compared with control cells. Besides, the microRNA-124 expression was higher in WT-DLX3 and MT-DLX3 but lower in Dlx3-shRNA. Moreover, the microRNA-124 expression level positively correlated with DLX3, negatively with osteoclastogenesis-related gene NFATc1. Our results indicate that DLX3 negatively regulates osteoclastic differentiation through microRNA-124, which is partially responsible for the increased bone density in TDO patient. DLX3 may be exploited for osteoclastogenesis regulator and potential therapeutic target of osteoporosis in future.
Assuntos
Proteínas de Homeodomínio/metabolismo , MicroRNAs/genética , Osteoclastos/metabolismo , Fatores de Transcrição/metabolismo , Animais , Diferenciação Celular , Hematopoese/genética , Camundongos , Osteoclastos/citologia , Deleção de SequênciaRESUMO
The aim of this study was to analyze the association between BMP2 (rs1884302) and DLX3 (rs2278163) gene polymorphisms and dental caries in primary and permanent dentitions. A total of 914 subjects were genotyped by the TaqMan methods: 176 caries-free children (with Decayed/Missing/Filled Teeth, DMFT = 0), 542 patients with dental caries in permanent dentition (DMFT ≥1), 83 caries-free children with primary teeth (with decayed/missing/filled teeth, dmft = 0), and 113 children with early childhood caries (ECC, dmft ≥1). There were no significant differences in allele/genotype frequencies between patients with caries in permanent dentition/ECC and caries-free children or between patients with very low (DMFT = 0-2), low (DMFT = 3-5), moderate (DMFT = 6-8), or high (DMFT ≥9) caries experience. Variability in BMP2 and DLX3 was not associated with caries in the Czech population.
Assuntos
Proteína Morfogenética Óssea 2/genética , Cárie Dentária/genética , Proteínas de Homeodomínio/genética , Polimorfismo de Nucleotídeo Único , Fatores de Transcrição/genética , Adolescente , Alelos , Criança , Pré-Escolar , República Tcheca , Índice CPO , Dentição Permanente , Feminino , Predisposição Genética para Doença , Genótipo , Humanos , Masculino , Índice de Gravidade de Doença , Dente DecíduoRESUMO
OBJECTIVE: To identify the molecular genetic aetiology of a family with autosomal dominant amelogenesis imperfecta (AI). SUBJECTS AND METHODS: DNA samples were collected from a six-generation family, and the candidate gene approach was used to screen for the enamelin (ENAM) gene. Whole-exome sequencing and linkage analysis with SNP array data identified linked regions, and candidate gene screening was performed. RESULTS: Mutational analysis revealed a mutation (c.561_562delCT and p.Tyr188Glnfs*13) in the DLX3 gene. After finding a recurrent DLX3 mutation, the clinical phenotype of the family members was re-examined. The proband's mother had pulp elongation in the third molars. The proband had not hair phenotype, but her cousin had curly hair at birth. CONCLUSIONS: In this study, we identified a recurrent 2-bp deletional DLX3 mutation in a new family. The clinical phenotype was the mildest one associated with the DLX3 mutations. These results will advance the understanding of the functional role of DLX3 in developmental processes.
Assuntos
Amelogênese Imperfeita/genética , Sequência de Bases , Proteínas de Homeodomínio/genética , Deleção de Sequência , Fatores de Transcrição/genética , Adulto , Amelogênese Imperfeita/diagnóstico por imagem , Criança , Análise Mutacional de DNA , Exoma , Feminino , Humanos , Masculino , Linhagem , Fenótipo , Radiografia DentáriaRESUMO
Tricho-dento-osseous (TDO) syndrome, an autosomal-dominant disorder, affects the morphological appearance of the tooth enamel, hair, and bone. Previous studies have confirmed that mutations in the DLX3 gene are responsible for TDO. In this study, we describe a Chinese patient with the typical traits of TDO - kinky hair, enamel hypoplasia, skull and jaw bones thickening, and sclerosis. Unfortunately, as a result of excessive attrition, we were unable to assess taurodontism. Examination of the tooth ground section showed a thin layer of enamel with no rods on the patient's tooth and abnormalities in Tomes' granular layer and the dentinal tubules. Scanning electron microscopy and energy-dispersive X-ray spectroscopy of the tooth enamel showed significant differences between the patient and the control individuals. A hair sample from the patient observed under a laser-scanning microscope showed longitudinal grooves in the hair shaft. Dual-energy X-ray absorptiometry measurement showed that the bone mineral density values of the patient's bones was much higher than normal. Finally, genetic analysis revealed a novel de novo missense mutation c.533A>G (p.Q178R) in the conserved homeodomain of the DLX3 gene. This DLX3 mutation is the sixth causative mutation for TDO to be identified so far.
Assuntos
Anormalidades Craniofaciais/genética , Hipoplasia do Esmalte Dentário/genética , Doenças do Cabelo/genética , Proteínas de Homeodomínio/genética , Mutação de Sentido Incorreto/genética , Fatores de Transcrição/genética , Absorciometria de Fóton , Adenina , Adulto , Densidade Óssea/fisiologia , China , Sequência Conservada/genética , Anormalidades Craniofaciais/patologia , Esmalte Dentário/anormalidades , Esmalte Dentário/ultraestrutura , Hipoplasia do Esmalte Dentário/patologia , Dentina/anormalidades , Éxons/genética , Feminino , Guanina , Cabelo/anormalidades , Cabelo/ultraestrutura , Doenças do Cabelo/patologia , Humanos , Imageamento Tridimensional , Íntrons/genética , Microscopia Confocal , Microscopia Eletrônica de Varredura , Espectrometria por Raios XRESUMO
BACKGROUND: The peripheral olfactory sensory system arises from morphologically identifiable structures called placodes. Placodes are relatively late developing structures, evident only well after the initiation of somitogenesis. Placodes are generally described as being induced from the ectoderm suggesting that their development is separate from the coordinated cell movements generating the central nervous system. RESULTS: With the advent of modern techniques it is possible to follow the development of the neurectoderm giving rise to the anterior neural tube, including the olfactory placodes. The cell movements giving rise to the optic cup are coordinated with those generating the olfactory placodes and adjacent telencephalon. The formation of the basal lamina separating the placode from the neural tube is coincident with the anterior migration of cranial neural crest. CONCLUSIONS: Olfactory placodes are transient morphological structures arising from a continuous sheet of neurectoderm that gives rise to the peripheral and central nervous system. This field of cells is specified at the end of gastrulation and not secondarily induced from ectoderm. The separation of olfactory placodes and telencephalon occurs through complex cell movements within the developing neural plate similar to that observed for the developing optic cup.
Assuntos
Placa Neural/embriologia , Tubo Neural/embriologia , Bulbo Olfatório/embriologia , Organogênese/fisiologia , Peixe-Zebra/embriologia , Animais , Placa Neural/citologia , Tubo Neural/citologia , Bulbo Olfatório/citologiaRESUMO
Protein kinase A (PKA), a serine/threonine kinase, regulates bone formation, and enhances Bone morphogenetic protein (BMP)-induced osteoblast differentiation. However, the mechanisms of how PKA controls the cellular response to BMP are not well known. We investigated the effects of modulating PKA activity during BMP2-induced osteoblast differentiation, and found that PKA regulates the function of Dlx3. Dlx3 plays crucial roles in osteoblast differentiation and it is expressed in most skeletal elements during development. We found that PKA activation increases BMP2-induced expression of Dlx3 protein, and enhances the protein stability, DNA binding, and transcriptional activity of Dlx3. In addition, PKA activation induces the phosphorylation of Dlx3 at consensus PKA phosphorylation target site(s). Lastly, substitution of serine 10 in Dlx3 to alanine significantly reduces, if not completely abolishes, the phosphorylation of Dlx3 and the regulation of Dlx3 function by PKA. These results suggest that Dlx3 is a novel target of PKA, and that PKA mediates BMP signaling during osteoblast differentiation, at least in part, by phosphorylating Dlx3 and modulating the protein stability and function of Dlx3.
Assuntos
Proteína Morfogenética Óssea 2/metabolismo , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Osteoblastos/fisiologia , Serina/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Substituição de Aminoácidos , Animais , Proteína Morfogenética Óssea 2/farmacologia , Diferenciação Celular , Linhagem Celular , Colforsina/farmacologia , Células HEK293 , Proteínas de Homeodomínio/química , Humanos , Isoquinolinas/farmacologia , Camundongos , Fosforilação , Estabilidade Proteica , Sulfonamidas/farmacologia , Fatores de Transcrição/químicaRESUMO
Dental follicle cells (DFCs) are dental stem/progenitor cells and the genuine precursors of alveolar osteoblasts and dental cementoblasts. A previous study showed that the transcription factor DLX3 (distal less homeobox 3) supports the osteogenic differentiation in DFCs via a positive feedback loop with the bone morghogenetic protein (BMP) 2. Until today, however, the control of this BMP2/DLX3 pathway by additional signaling pathways remains elusive. Previous studies also suggested that the NOTCH signaling pathway plays a role in the osteogenic differentiation of DFCs. In this study we showed that DLX3 overexpression and the initiation of the osteogenic differentiation by BMP2 or dexamethasone induced the NOTCH signaling pathway in DFCs. However, the induction of NOTCH-signaling impaired not only the osteogenic differentiation (ALP activity and mineralized nodules) but also the expression of the transcription factor DLX3 and the activation of the BMP-signaling pathway. So, NOTCH signaling plays a regulatory role for the osteogenic differentiation of DFCs. In conclusion, results of our study suggest that the NOTCH-signaling pathway, which is activated during the osteogenic differentiation of DFCs, regulates the BMP2/DLX3 directed differentiation of DFCs via a negative feed-back loop.
Assuntos
Proteína Morfogenética Óssea 2/metabolismo , Proteínas de Homeodomínio/metabolismo , Odontogênese/fisiologia , Osteoblastos/citologia , Osteoblastos/fisiologia , Dente/citologia , Dente/fisiologia , Fatores de Transcrição/metabolismo , Diferenciação Celular , Células Cultivadas , Células do Tecido Conjuntivo/citologia , Células do Tecido Conjuntivo/fisiologia , Humanos , Osteogênese/fisiologia , Receptor Notch1 , Transdução de Sinais/fisiologiaRESUMO
We report on a 22-year-old woman with features of osteogenesis imperfecta (OI), tricho-dento-osseous (TDO) syndrome and intellectual disability. Whole genome oligonucleotide microarray analysis revealed a copy number gain of 3 Mb in 7q32.3-q33 and a loss of 3.4 Mb in 17q21.33-q22. FISH analysis showed that the third copy of 7q32 was inserted into the long arm of one chromosome 17, exactly in the region 17q21.33-q22 that was deleted. The maternal uncle presented with clinical features similar to the proposita and had the same chromosomal anomalies. The mother of the proposita and two other family members were balanced carriers of this rearrangement, interpreted as an interchromosomal reciprocal insertion. Reciprocal insertion/four-break rearrangement is a very rare chromosomal event. The deleted region on chromosome 17 contains 39 genes, including COL1A1 and DLX3 involved in OI and TDO syndrome respectively. The CACNA1G gene on the deleted segment of chromosome 17 may be a good candidate gene to explain the intellectual impairment. © 2013 Wiley Periodicals, Inc.
Assuntos
Anormalidades Craniofaciais/genética , Hipoplasia do Esmalte Dentário/genética , Doenças do Cabelo/genética , Deficiência Intelectual/genética , Osteogênese Imperfeita/genética , Duplicação Cromossômica , Cromossomos Humanos Par 17 , Cromossomos Humanos Par 7 , Colágeno Tipo I/genética , Cadeia alfa 1 do Colágeno Tipo I , Hibridização Genômica Comparativa , Anormalidades Craniofaciais/diagnóstico , Hipoplasia do Esmalte Dentário/diagnóstico , Feminino , Doenças do Cabelo/diagnóstico , Proteínas de Homeodomínio/genética , Humanos , Hibridização in Situ Fluorescente , Deficiência Intelectual/diagnóstico , Masculino , Pessoa de Meia-Idade , Mutagênese Insercional , Osteogênese Imperfeita/diagnóstico , Linhagem , Fenótipo , Deleção de Sequência , Síndrome , Fatores de Transcrição/genética , Adulto JovemRESUMO
Non syndromic cleft lip and/or palate (NSCLP) is a complex congenital anomaly with varying incidence among patients of different geographical origins. Multiple contributing factors are known to trigger the cleft formation. There are several genes involved in the aetiology of NSCLP and they are different in different populations. The genetic components of clefts that underlie the susceptibility to respond to the environment still remain unclear. In this study, five microsatellite polymorphisms from five candidate genes were employed to analyze the association between these genes and NSCLP in 83 patients and 90 controls. Genotyping was performed by separating and visualizing the fluorescently-labeled polymerase chain reaction (PCR) products. The association of the five microsatellite polymorphisms with NSCLP was tested by using the CLUMP v1.9 program that uses the Monte Carlo method. The genotypic distribution is in Hardy-Weinberg equilibrium in the control group for only the MSX1 and DLX3 genes. The RARA microsatellite was significantly associated with NSCLP. Our results suggest that the RARA gene is involved in pathogenesis of cleft lip and palate in South Indians.
RESUMO
Uterine leiomyomas (ULs) are benign solid tumors arising from the uterine myometrium. They are the most common pelvic tumors among females of reproductive age. Despite the universal prevalence of ULs and its huge impact on women's lives, the exact etiology and pathophysiologic mechanisms have not been fully understood. Numerous studies indicate that genetic factors play a crucial role in ULs development. This study aims to identify the probable genetic causes of ULs in a consanguineous Iranian family. Whole-exome sequencing (WES) on five family members with ULs revealed a likely pathogenic missense variant encoding for Y88C in the transactivation (TA) domain of DLX3 gene (c.263A > G; p.Y88C). Sanger sequencing of a total of 9 affected and non-affected family members indicated a segregation with disease with autosomal dominant inheritance. Moreover, targeted Sanger sequencing on 32 additional non-related patients with ULs showed none was heterozygous for this variant. MutPred2 predicted the pathogenicity of candidate variant by both phosphorylation and sulfation loss as actionable hypotheses. Project HOPE revealed that the identified variant residue is smaller and more hydrophobic comparing to the wild-type residue. I-TASSER and UCSF Chimera were also used for modeling and visualizing the predicted variant, respectively. This WES analysis is the first to report a variant in DLX3 variation associated with ULs pathogenicity in Iranian population highlighting the effectiveness of WES as a strong diagnostic method. However, further functional studies on this variant are needed to confirm the potential pathogenicity of this mutation.
Assuntos
Aborto Espontâneo , Leiomioma , Feminino , Humanos , Gravidez , Consanguinidade , Irã (Geográfico) , Leiomioma/genética , Mutação , Mutação de Sentido Incorreto , LinhagemRESUMO
OBJECTIVES: To identify DLX3 variants in a Chinese family with typical clinical manifestations of tricho-dento-osseous syndrome (TDO). DESIGN: Sanger sequencing was performed to detect DLX3 variants in the TDO family. Three-dimensional laser scanning microscopy, bioinformatic and conformational analyses were employed to explore the phenotypic characterization and the functional impact. RESULTS: We identified a novel heterozygous variant in the DLX3 gene (c.534G>C; p.Gln178His). Familial co-segregation verified an autosomal dominant inheritance pattern. Bioinformatic prediction demonstrated the deleterious effects of the variant, and DLX3 structure changes suggested the corresponding functional impairments. CONCLUSIONS: We identified a variant in the DLX3 gene in an integrated family of Han nationality for the first time. This study expands the variant spectrum of DLX3 and phenotype spectrum of TDO syndrome.
Assuntos
Hipoplasia do Esmalte Dentário , Doenças do Cabelo , Proteínas de Homeodomínio , Fatores de Transcrição , Anormalidades Craniofaciais , Hipoplasia do Esmalte Dentário/genética , Doenças do Cabelo/genética , Proteínas de Homeodomínio/genética , Humanos , Linhagem , Fatores de Transcrição/genéticaRESUMO
MicroRNAs play an important role in myogenic differentiation, they bind to target genes and regulate muscle formation. We previously found that miR-9-5p, which is related to bone formation, was increased over time during the process of myogenic differentiation. However, the mechanism by which miR-9-5p regulates myogenic differentiation remains largely unknown. In the present study, we first examined myotube formation and miR-9-5p, myogenesis-related genes including Dlx3, Myod1, Mef2c, Desmin, MyoG and Myf5 expression under myogenic induction. Then, we detected the expression of myogenic transcription factors after overexpression or knockdown of miR-9-5p or Dlx3 in the mouse premyoblast cell line C2C12 by qPCR, western blot and myotube formation under myogenic induction. A luciferase assay was performed to confirm the regulatory relationships between not only miR-9-5p and Dlx3 but also Dlx3 and its downstream gene, Myf5, which is an essential transcription factor of myogenic differentiation. The results showed that miR-9-5p promoted myogenic differentiation by increasing myogenic transcription factor expression and promoting myotube formation, but Dlx3 exerted the opposite effect. Moreover, the luciferase assay showed that miR-9-5p bound to the 3'UTR of Dlx3 and downregulated Dlx3 expression. Dlx3 in turn suppressed Myf5 expression by binding to the Myf5 promoter, ultimately inhibiting the process of myogenic differentiation. In conclusion, the miR-9-5p/Dlx3/Myf5 axis is a novel pathway for the regulation of myogenic differentiation, and can be a potential target to treat the diseases related to muscle dysfunction.
Assuntos
MicroRNAs , Camundongos , Animais , MicroRNAs/genética , Diferenciação Celular/genética , Linhagem Celular , Fatores de Transcrição/genética , Desenvolvimento Muscular/genética , Fator Regulador Miogênico 5/genéticaRESUMO
Lymphatic enhancer factor 1 (Lef1) and distal-less homeobox 3 (Dlx3) are the transcription factors involved in regulating hair follicle development in mice, goats, and other animals. Their deletion can lead to hair follicle deficiency. In this study, hematoxylin−eosin staining (HE), real-time quantitative PCR (RT-qPCR), immunohistochemistry, and immunofluorescence were used to analyze the expression, location, and biological functions of Lef1 and Dlx3 in the lateral skin of Gansu Alpine Merino aged 1, 30, 60, and 90 days. The results revealed that the number of hair follicles decreased with age and was significantly higher at 1 day than in the other three age groups (p < 0.05). The mRNA levels of Lef1 and Dlx3 in the skin of 30-day old Gansu Alpine Merino were significantly higher than those in the other three age groups (p < 0.05). Protein expression of Lef1 and Dlx3 was lowest at 1 day (p < 0.05) and peaked at 60 days. Lef1 and Dlx3 exhibited a high density and strong positive expression in the dermal papillae; additionally, Dlx3 exhibited a high density and strong positive expression in the inner and outer root sheaths. Collectively, Lef1 and Dlx3 may facilitate the maturation of secondary hair follicles, which is mainly achieved through the dermal papillae and inner and outer root sheaths.
Assuntos
Folículo Piloso , Proteínas de Homeodomínio , Fator 1 de Ligação ao Facilitador Linfoide , Fatores de Transcrição , Animais , Proteínas de Homeodomínio/genética , Fator 1 de Ligação ao Facilitador Linfoide/genética , Sequências Reguladoras de Ácido Nucleico , Carneiro Doméstico/genética , Pele/metabolismo , Fatores de Transcrição/genéticaRESUMO
Osteoporosis is a complex multifactorial disorder linked to various risk factors and medical conditions. Bone marrow-derived mesenchymal stem cell (BMSC) dysfunction potentially plays a critical role in osteoporosis pathogenesis. Herein, the study identified that miR-4739 was upregulated in BMSC cultures harvested from osteoporotic subjects. BMSCs were isolated from normal and osteoporotic bone marrow tissues and identified for their osteogenic differentiation potential. In osteoporotic BMSCs, miR-4739 overexpression significantly inhibited cell viability, osteoblast differentiation, mineralized nodule formation, and heterotopic bone formation, whereas miR-4739 inhibition exerted opposite effects. Through direct binding, miR-4739 inhibited distal-less homeobox 3 (DLX3) expression. In osteoporotic BMSCs, DLX3 knockdown also inhibited BMSC viability and osteogenic differentiation. Moreover, DLX3 knockdown partially attenuated the effects of miR-4739 inhibition upon BMSCs. Altogether, the miR-4739/DLX3 axis modulates the capacity of BMSCs to differentiate into osteoblasts, which potentially plays a role in osteoporosis pathogenesis. The in vivo and clinical functions of the miR-4739/DLX3 axis require further investigation.