Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 405
Filtrar
1.
Nat Commun ; 10(1): 3993, 2019 09 05.
Artigo em Inglês | MEDLINE | ID: mdl-31488837

RESUMO

Planar cell polarity (PCP) plays crucial roles in developmental processes such as gastrulation, neural tube closure and hearing. Wnt pathway mutants are often classified as PCP mutants due to similarities between their phenotypes. Here, we show that in the zebrafish lateral line, disruptions of the PCP and Wnt pathways have differential effects on hair cell orientations. While mutations in the PCP genes vangl2 and scrib cause random orientations of hair cells, mutations in wnt11f1, gpc4 and fzd7a/b induce hair cells to adopt a concentric pattern. This concentric pattern is not caused by defects in PCP but is due to misaligned support cells. The molecular basis of the support cell defect is unknown but we demonstrate that the PCP and Wnt pathways work in parallel to establish proper hair cell orientation. Consequently, hair cell orientation defects are not solely explained by defects in PCP signaling, and some hair cell phenotypes warrant re-evaluation.


Assuntos
Polaridade Celular/genética , Polaridade Celular/fisiologia , Células Ciliadas Auditivas/metabolismo , Via de Sinalização Wnt/genética , Via de Sinalização Wnt/fisiologia , Peixe-Zebra/genética , Animais , Regulação da Expressão Gênica no Desenvolvimento , Proteoglicanas de Heparan Sulfato/genética , Proteínas de Membrana/genética , Morfogênese/genética , Morfogênese/fisiologia , Mutação , Defeitos do Tubo Neural/genética , Neurulação/genética , Receptores de Superfície Celular/genética , Proteína Wnt1/genética , Proteínas de Peixe-Zebra/genética
2.
Med Arch ; 73(3): 144-148, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31402800

RESUMO

Introduction: Previous research found that diabetes mellitus capable to aggravate osteoarthritis disease. In brief, the hyperglycemia condition in diabetes mellitus has an impact on protein glycation of all joint components, including molecule, such as perlecan. The protein expression of perlecan reflects the presence amount of perlecan in the matrix of articular cartilage. However, the impact of hyperglycemia on articular perlecan has not been explained. Moreover, the role of perlecan as a mechanotransducer for chondrocytes in type 1 Diabetes mellitus remains unclear. Aim: This research aims to analyze the effect of hyperglycemia in type 1 Diabetes mellitus to the mRNA level and protein expression of perlecan. Methods: Thirty-five adult male rats were divided into seven groups, such as three groups of rat model with anterior cruciate ligament transection (ACLT) at right knee (ACLT1, ACLT2, ACLT3); three groups of rats with ACLT at right knee which followed by Streptozotocin injection for diabetic mice model (DM1, DM2, DM3); and control group (N). Rat sacrificed at the third week, fourth week, and sixth week after two months of maintenance. The mRNA level and protein expression were analyzed by using PCR and Western blot. All of data was analyzed by ANOVA. Results: Protein expression of perlecan in ACLT mice with diabetes mellitus (DM1, DM2, DM3 group) was gradually decreased according to the increased hyperglycemia duration. Whilst, protein expression of perlecan within ACLT mice without diabetes mellitus (ACLT1, ACLT2, ACLT3 group) was increased. The similar result also demonstrated by the mRNA level of perlecan. Group of DM1, DM2, DM3 exhibited decreased mRNA level of perlecan over the hyperglycemia duration. While, ACLT1, ACLT2, and ACLT3 group had a gradually increased of perlecan mRNA level. Conclusion: Hyperglycemia on osteoarthritic condition decreased mRNA level and protein expression of perlecan which increased the severity of osteoarthritis disease.


Assuntos
Diabetes Mellitus Experimental/sangue , Diabetes Mellitus Tipo 1/sangue , Proteoglicanas de Heparan Sulfato/genética , Proteoglicanas de Heparan Sulfato/metabolismo , Hiperglicemia/sangue , Osteoartrite/metabolismo , Animais , Glicemia , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Tipo 1/complicações , Modelos Animais de Doenças , Hiperglicemia/etiologia , Masculino , Osteoartrite/complicações , Osteoartrite/genética , RNA Mensageiro/metabolismo , Ratos
3.
J Mol Histol ; 50(3): 285-294, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30993430

RESUMO

The aim of this study was to ascertain whether, like many cell types in cartilaginous tissues if type XI collagen was a pericellular component of annulus fibrosus (AF) cells and chondrocytes. Fine fibrillar networks were visualised which were perlecan, HS (MAb 10E4) and type XI collagen positive. Heparitinase-III pre-digestion abolished the type XI collagen and 10E4 localisation in these fibrillar assemblies demonstrating a putative HS mediated interaction which localised the type XI collagen. Type XI collagen was confirmed to be present in the Heparitinase III treated AF monolayer media samples by immunoblotting. Heparitinase-III generated ΔHS stub epitopes throughout these fibrillar networks strongly visualised by MAb 3-G-10. Monolayers of murine hip articular chondrocytes from C57BL/6 and Hspg2 exon 3 null mice also displayed pericellular perlecan localisations, however type XI collagen was only evident in the Wild type mice. Perlecan was also immunolocalised in control and murine knee articular cartilage from the two mouse genotypes subjected to a medial meniscal destabilisation procedure which induces OA. This resulted in a severe depletion of perlecan levels particularly in the perlecan exon 3 null mice and was consistent with OA representing a disease of the pericellular matrix. A model was prepared to explain these observations between the NPP type XI collagen domain and HS chains of perlecan domain-I in the pericellular matrix of AF cells which likely contributed to cellular communication, tissue stabilization and the regulation of extracellular matrix homeostasis.


Assuntos
Anel Fibroso/efeitos dos fármacos , Colágeno Tipo XI/genética , Proteoglicanas de Heparan Sulfato/genética , Animais , Anel Fibroso/metabolismo , Anel Fibroso/patologia , Cartilagem Articular/crescimento & desenvolvimento , Cartilagem Articular/metabolismo , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Éxons/genética , Matriz Extracelular/genética , Proteínas da Matriz Extracelular/genética , Homeostase/genética , Disco Intervertebral/crescimento & desenvolvimento , Disco Intervertebral/metabolismo , Camundongos , Polissacarídeo-Liase/farmacologia
4.
BMC Musculoskelet Disord ; 20(1): 24, 2019 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-30646882

RESUMO

BACKGROUND: Rare variants of HSPG2 have recently been reported to function as a potential contributor to the susceptibility of adolescent idiopathic scoliosis (AIS) in the Caucasians. A replication study in the different population is warranted to validate the role of HSPG2 in AIS. The aim of this study was to determine the association between HSPG2 and AIS in the Chinese patients and to further investigate its influence on the phenotype of the patients. METHODS: SNVs p.Asn786Ser of HSPG2 was genotyped in 1752 patients and 1584 normal controls using multiple ligase detection reactions. The mRNA expression of HSPG2 in the paraspinal muscles was quantified for 90 patients and 26 controls. The The Student's t test was used to analyze the inter-group comparison of the HSPG2 expression. The relationship between the HSPG2 expression and the curve magnitude of the patients was analyzed by the Pearson correlation analysis. RESULTS: No case of mutation in the reported SNV p.Asn786Ser of HSPG2 was found in our cohort. The mRNA expression of HSPG2 in patients was comparable with that in the controls (0.0016 ± 0.0013 vs. 0.0019 ± 0.0012, p = 0.29). 42 patients with curve magnitude > 60 degrees were assigned to the severe curve group. The other 58 patients were assigned to the moderate curve group. These two groups were found to have comparable HSPG2 expression (0.0015 ± 0.0011 vs. 0.0017 ± 0.0014, p = 0.57). And there was no remarkable correlation between the expression level of HSPG2 and the curve severity (r = 0.131, p = 0.71). CONCLUSIONS: HSPG2 gene was not associated with the susceptibility or the phenotypes of AIS in the Chinese population. The whole HSPG2 gene can be sequenced in more AIS patients to identify potentially causative mutations.


Assuntos
Grupo com Ancestrais do Continente Asiático/genética , Predisposição Genética para Doença , Proteoglicanas de Heparan Sulfato/genética , Escoliose/genética , Adolescente , Adulto , Estudos de Casos e Controles , Criança , Estudos de Coortes , Análise Mutacional de DNA , Feminino , Estudo de Associação Genômica Ampla/estatística & dados numéricos , Glicosídeos/genética , Humanos , Polimorfismo de Nucleotídeo Único , Esteróis , Adulto Jovem
5.
Matrix Biol ; 75-76: 220-259, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-29128506

RESUMO

Extracellular matrix is a highly dynamic macromolecular network. Proteoglycans are major components of extracellular matrix playing key roles in its structural organization and cell signaling contributing to the control of numerous normal and pathological processes. As multifunctional molecules, proteoglycans participate in various cell functions during morphogenesis, wound healing, inflammation and tumorigenesis. Their interactions with matrix effectors, cell surface receptors and enzymes enable them with unique properties. In malignancy, extensive remodeling of tumor stroma is associated with marked alterations in proteoglycans' expression and structural variability. Proteoglycans exert diverse functions in tumor stroma in a cell-specific and context-specific manner and they mainly contribute to the formation of a permissive provisional matrix for tumor growth affecting tissue organization, cell-cell and cell-matrix interactions and tumor cell signaling. Proteoglycans also modulate cancer cell phenotype and properties, the development of drug resistance and tumor stroma angiogenesis. This review summarizes the proteoglycans remodeling and their novel biological roles in malignancies with particular emphasis to the underlying molecular mechanisms.


Assuntos
Matriz Extracelular/genética , Glicosaminoglicanos/genética , Proteoglicanas de Heparan Sulfato/genética , Neovascularização Patológica/genética , Proteínas da Matriz Extracelular/química , Proteínas da Matriz Extracelular/genética , Glicosaminoglicanos/química , Proteoglicanas de Heparan Sulfato/química , Humanos , Morfogênese/genética , Neoplasias/genética , Neoplasias/patologia , Neovascularização Patológica/patologia , Receptores de Superfície Celular/química , Receptores de Superfície Celular/genética , Transdução de Sinais , Cicatrização/genética
6.
Biochem J ; 476(2): 225-243, 2019 01 18.
Artigo em Inglês | MEDLINE | ID: mdl-30563944

RESUMO

Heparan sulfate (HS) regulates diverse cell signalling events in intervertebral disc development and homeostasis. The aim of the present study was to investigate the effect of ablation of perlecan HS/CS on murine intervertebral disc development. Genetic models carrying mutations in genes encoding HS biosynthetic enzymes have identified multiple roles for HS in tissue homeostasis. In the present study, we utilised an Hspg2 exon 3 null HS/CS-deficient mouse to assess the role of perlecan HS in disc cell regulation. HS makes many important contributions to growth factor sequestration, stabilisation/delivery, and activation of receptors directing cellular proliferation, differentiation, and assembly of extracellular matrix. Perlecan HS/CS-mediated interactions promote extracellular matrix assembly/stabilisation and tissue functional properties, and thus, removal of perlecan HS/CS should affect extracellular matrix function and homeostasis. Hspg2 exon 3 null intervertebral discs accumulated significantly greater glycosaminoglycan in the nucleus pulposus, annulus fibrosus, and vertebral growth plates than C57BL/6 wild-type (WT) I intervertebral discs. Proliferation of intervertebral disc progenitor cells was significantly higher in Hspg2 exon 3 null intervertebral discs, and these cells became hypertrophic by 12 weeks of age and were prominent in the vertebral growth plates but had a disorganised organisation. C57BL/6 WT vertebral growth plates contained regular columnar growth plate chondrocytes. Exostosis-like, ectopic bone formation occurred in Hspg2 exon 3 null intervertebral discs, and differences were evident in disc cell maturation and in matrix deposition in this genotype, indicating that perlecan HS/CS chains had cell and matrix interactive properties which repressively maintained tissue homeostasis in the adult intervertebral disc.


Assuntos
Proliferação de Células , Éxons , Glicosaminoglicanos/metabolismo , Lâmina de Crescimento/metabolismo , Proteoglicanas de Heparan Sulfato/metabolismo , Núcleo Pulposo/metabolismo , Animais , Condrócitos/metabolismo , Condrócitos/patologia , Matriz Extracelular/genética , Matriz Extracelular/metabolismo , Matriz Extracelular/patologia , Glicosaminoglicanos/genética , Lâmina de Crescimento/patologia , Proteoglicanas de Heparan Sulfato/genética , Hipertrofia , Camundongos , Camundongos Mutantes , Núcleo Pulposo/patologia , Células-Tronco/metabolismo , Células-Tronco/patologia
7.
Invest Ophthalmol Vis Sci ; 59(13): 5589-5598, 2018 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-30480706

RESUMO

Purpose: To determine whether (1) the in vitro expression of epithelial basement membrane components nidogen-1, nidogen-2, and perlecan by keratocytes, corneal fibroblasts, and myofibroblasts is modulated by cytokines/growth factors, and (2) perlecan protein is produced by stromal cells after photorefractive keratectomy. Methods: Marker-verified rabbit keratocytes, corneal fibroblasts, myofibroblasts were stimulated with TGF-ß1, IL-1α, IL-1ß, TGF-ß3, platelet-derived growth factor (PDGF)-AA, or PDGF-AB. Real-time quantitative RT-PCR was used to detect expression of nidogen-1, nidogen-2, and perlecan mRNAs. Western blotting evaluated changes in protein expression. Immunohistochemistry was performed on rabbit corneas for perlecan, alpha-smooth muscle actin, keratocan, vimentin, and CD45 at time points from 1 day to 1 month after photorefractive keratectomy (PRK). Results: IL-1α or -1ß significantly upregulated perlecan mRNA expression in keratocytes. TGF-ß1 or -ß3 markedly downregulated nidogen-1 or -2 mRNA expression in keratocytes. None of these cytokines had significant effects on nidogen-1, -2, or perlecan mRNA expression in corneal fibroblasts or myofibroblasts. IL-1α significantly upregulated, while TGF-ß1 significantly downregulated, perlecan protein expression in keratocytes. Perlecan protein expression was upregulated in anterior stromal cells at 1 and 2 days after -4.5 or -9 diopters (D) PRK, but the subepithelial localization of perlecan became disrupted at 7 days and later time points in -9-D PRK corneas when myofibroblasts populated the anterior stroma. Conclusions: IL-1 and TGF-ß1 have opposing effects on perlecan and nidogen expression by keratocytes in vitro. Proximate participation of keratocytes is likely needed to regenerate normal epithelial basement membrane after corneal injury.


Assuntos
Ceratócitos da Córnea/efeitos dos fármacos , Substância Própria/efeitos dos fármacos , Regulação da Expressão Gênica/fisiologia , Proteoglicanas de Heparan Sulfato/genética , Interleucina-1/farmacologia , Glicoproteínas de Membrana/genética , Fator de Crescimento Transformador beta/farmacologia , Animais , Membrana Basal/metabolismo , Western Blotting , Ceratócitos da Córnea/metabolismo , Substância Própria/metabolismo , Proteoglicanas de Heparan Sulfato/metabolismo , Técnicas Imunoenzimáticas , Interleucina-1alfa/farmacologia , Interleucina-1beta/farmacologia , Glicoproteínas de Membrana/metabolismo , RNA Mensageiro/genética , Coelhos , Reação em Cadeia da Polimerase em Tempo Real , Fator de Crescimento Transformador beta1/farmacologia , Fator de Crescimento Transformador beta3/farmacologia
8.
Eur Rev Med Pharmacol Sci ; 22(20): 6853-6863, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-30402850

RESUMO

OBJECTIVE: Perlecan, which is also called heparan sulfate proteoglycan 2 (HSPG2), is a protein encoded by the HSPG2 gene that maps to 1p36.12 in the human genome. In this study, we assessed the independent prognostic value of HSPG2 in terms of overall survival (OS) and recurrence-free survival (RFS) in patients with LGG. PATIENTS AND METHODS: A retrospective study was conducted by using data in the Cancer Genome Atlas-Low Grade Glioma (TCGA-LGG). RESULTS: Increased HSPG2 expression was an independent prognostic indicator of poor OS in oligoastrocytoma (HR: 1.644, 95% CI: 1.116-2.423, p = 0.012) and in oligodendroglioma (HR: 1.459, 95% CI: 1.138-1.871, p = 0.003). In addition, increased HSPG2 expression independently predicted poor RFS in oligodendroglioma (HR: 1.402, 95% CI: 1.110-1.770, p = 0.005). Furthermore, we observed that high HSPG2 expression was associated with significantly shorter OS and RFS in oligodendroglioma, no matter the patients received radiotherapy or not. Using copy number alterations (CNAs) and DNA methylation data in TCGA-LGG, we found that DNA copy deletion was generally associated with decreased HSPG2 expression. Regression analysis suggested a weak negative correlation between HSPG2 expression and HSPG2 DNA methylation (Pearson's r = -0.388). CONCLUSIONS: Increased HSPG2 expression could independently predict poor OS in oligoastrocytoma and oligodendroglioma and also independently predicted poor RFS in oligodendroglioma. Its expression is modulated by both DNA copy number and DNA methylation in oligodendroglioma.


Assuntos
Metilação de DNA , Proteoglicanas de Heparan Sulfato/genética , Oligodendroglioma/genética , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Estudos Retrospectivos
9.
Front Immunol ; 9: 2191, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30327649

RESUMO

Objective: The mechanisms that lead to endothelial cell (EC) injury and propagate the vasculopathy in Systemic Sclerosis (SSc) are not well understood. Using single cell RNA sequencing (scRNA-seq), our goal was to identify EC markers and signature pathways associated with vascular injury in SSc skin. Methods: We implemented single cell sorting and subsequent RNA sequencing of cells isolated from SSc and healthy control skin. We used t-distributed stochastic neighbor embedding (t-SNE) to identify the various cell types. We performed pathway analysis using Gene Set Enrichment Analysis (GSEA) and Ingenuity Pathway Analysis (IPA). Finally, we independently verified distinct markers using immunohistochemistry on skin biopsies and qPCR in primary ECs from SSc and healthy skin. Results: By combining the t-SNE analysis with the expression of known EC markers, we positively identified ECs among the sorted cells. Subsequently, we examined the differential expression profile between the ECs from healthy and SSc skin. Using GSEA and IPA analysis, we demonstrated that the SSc endothelial cell expression profile is enriched in processes associated with extracellular matrix generation, negative regulation of angiogenesis and epithelial-to-mesenchymal transition. Two of the top differentially expressed genes, HSPG2 and APLNR, were independently verified using immunohistochemistry staining and real-time qPCR analysis. Conclusion: ScRNA-seq, differential gene expression and pathway analysis revealed that ECs from SSc patients show a discrete pattern of gene expression associated with vascular injury and activation, extracellular matrix generation and negative regulation of angiogenesis. HSPG2 and APLNR were identified as two of the top markers of EC injury in SSc.


Assuntos
Receptores de Apelina/genética , Receptores de Apelina/metabolismo , Células Endoteliais/metabolismo , Proteoglicanas de Heparan Sulfato/genética , Proteoglicanas de Heparan Sulfato/metabolismo , Escleroderma Sistêmico/genética , Escleroderma Sistêmico/metabolismo , Sequência de Bases , Biomarcadores/metabolismo , Biópsia , Ativação do Complemento , Transição Epitelial-Mesenquimal/genética , Matriz Extracelular/genética , Matriz Extracelular/metabolismo , Expressão Gênica , Humanos , Imuno-Histoquímica , Metaloproteinases da Matriz/metabolismo , Neovascularização Patológica/metabolismo , Escleroderma Sistêmico/patologia , Análise de Sequência de RNA , Lesões do Sistema Vascular/patologia
10.
Cell Physiol Biochem ; 50(1): 41-51, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30278461

RESUMO

BACKGROUND/AIMS: Maternally expressed gene 3 (MEG3) is an imprinted gene with maternal expression, which may function as a tumor suppressor by inhibiting angiogenesis. To identify the prognostic value of MEG3 in breast cancer, systematic analysis was performed in this study. METHODS: To evaluate gene alteration during breast carcinogenesis, we explored MEG3 expression using the Serial Analysis of Gene Expression Genie suite and Oncomine analysis. The prognostic roles of MEG3 in breast cancer were investigated using the PrognoScan database. The heat map and methylation status of MEG3 were determined using the UCSC Genome Browser. RESULTS: We found that MEG3 was more frequently downregulated in breast cancer than in normal tissues and this correlated with prognosis. However, estrogen receptor and progesterone receptor status were found to be positively correlated with MEG3 expression. Conversely, basal-like status, triple-negative breast cancer status, and Scarff Bloom & Richardson grade criterion were negatively correlated with MEG3 expression. Following data mining in multiple big data databases, we confirmed a positive correlation between MEG3 and heparan sulfate proteoglycan 2 (HSPG2) expression in breast cancer tissues. CONCLUSION: MEG3 could be adopted as a marker to predict the prognosis of breast cancer with HSPG2. However, large-scale and comprehensive research is needed to clarify our results.


Assuntos
Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/patologia , Biologia Computacional/métodos , RNA Longo não Codificante/metabolismo , Adulto , Biomarcadores Tumorais/genética , Neoplasias da Mama/metabolismo , Neoplasias da Mama/mortalidade , Metilação de DNA , Bases de Dados Factuais , Feminino , Regulação Neoplásica da Expressão Gênica , Proteoglicanas de Heparan Sulfato/genética , Proteoglicanas de Heparan Sulfato/metabolismo , Humanos , Pessoa de Meia-Idade , Prognóstico , RNA Longo não Codificante/genética , RNA Mensageiro/metabolismo , Análise de Sobrevida , Neoplasias de Mama Triplo Negativas/diagnóstico , Neoplasias de Mama Triplo Negativas/genética , Neoplasias de Mama Triplo Negativas/patologia
11.
Mol Ther ; 26(10): 2418-2430, 2018 10 03.
Artigo em Inglês | MEDLINE | ID: mdl-30057240

RESUMO

The present study was designed to characterize transduction of non-human primate brain and spinal cord with a modified adeno-associated virus serotype 2, incapable of binding to the heparan sulfate proteoglycan receptor, referred to as AAV2-HBKO. AAV2-HBKO was infused into the thalamus, intracerebroventricularly or via a combination of both intracerebroventricular and thalamic delivery. Thalamic injection of this modified vector encoding GFP resulted in widespread CNS transduction that included neurons in deep cortical layers, deep cerebellar nuclei, several subcortical regions, and motor neuron transduction in the spinal cord indicative of robust bidirectional axonal transport. Intracerebroventricular delivery similarly resulted in widespread cortical transduction, with one striking distinction that oligodendrocytes within superficial layers of the cortex were the primary cell type transduced. Robust motor neuron transduction was also observed in all levels of the spinal cord. The combination of thalamic and intracerebroventricular delivery resulted in transduction of oligodendrocytes in superficial cortical layers and neurons in deeper cortical layers. Several subcortical regions were also transduced. Our data demonstrate that AAV2-HBKO is a powerful vector for the potential treatment of a wide number of neurological disorders, and highlight that delivery route can significantly impact cellular tropism and pattern of CNS transduction.


Assuntos
Terapia Genética , Vetores Genéticos/efeitos adversos , Neurônios/efeitos dos fármacos , Parvovirinae/genética , Medula Espinal/efeitos dos fármacos , Animais , Transporte Axonal/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Encéfalo/patologia , Proteínas do Capsídeo/administração & dosagem , Proteínas do Capsídeo/genética , Sistema Nervoso Central/efeitos dos fármacos , Sistema Nervoso Central/patologia , Modelos Animais de Doenças , Vetores Genéticos/administração & dosagem , Vetores Genéticos/genética , Proteoglicanas de Heparan Sulfato/administração & dosagem , Proteoglicanas de Heparan Sulfato/genética , Humanos , Infusões Intraventriculares , Neurônios Motores/efeitos dos fármacos , Neurônios/patologia , Primatas , Medula Espinal/patologia , Tálamo/efeitos dos fármacos
12.
Mol Med Rep ; 18(2): 1761-1765, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29901129

RESUMO

Schwartz-Jampel syndrome type 1 (SJS1) is a rare autosomal recessive disease caused by mutations in the gene heparan sulfate proteoglycan 2 (HSPG2; also known as basement membrane­specific heparin sulfate). In the present study, a 10­year­old female SJS1 proband from a Chinese family, who was diagnosed by X­ray and physical examination, was recruited. The key clinical features of the patient with SJS1 included short stature, joint contractures, pigeon breast, and myotonia that led to progressive stiffness of the face and limbs; barely discernible kyphosis was also noted. Genetic testing using whole exome sequencing and Sanger sequencing was performed for the proband and family members. A total of 2 novel mutations (c.8788G>A; p.Glu2930Lys and c.11671+5G>A) in the HSPG2 gene were identified in the proband. The family members harboring 1 heterozygous mutation in HSPG2 did not exhibit any skeletal abnormalities. The results of the present study suggested that the compound heterozygous mutations in HSPG2 may be responsible the induction of SJS1, and demonstrated the genotype­phenotype associations between mutations in the HSPG2 gene and clinical characteristics of SJS1.


Assuntos
Estudos de Associação Genética , Proteoglicanas de Heparan Sulfato/genética , Osteocondrodisplasias/genética , Criança , China/epidemiologia , Feminino , Heterozigoto , Humanos , Mutação , Osteocondrodisplasias/diagnóstico , Osteocondrodisplasias/epidemiologia , Osteocondrodisplasias/patologia , Linhagem
13.
J Biol Chem ; 293(31): 12137-12148, 2018 08 03.
Artigo em Inglês | MEDLINE | ID: mdl-29921586

RESUMO

Regulation of autophagy by proteolytically cleaved fragments of heparan sulfate proteoglycans is a novel and current research focus in tumor biology. Endorepellin is the C-terminal angiostatic fragment of the heparan sulfate proteoglycan perlecan and induces autophagy in endothelial cells. To further investigate this property, we used NanoString, a digital PCR platform for measuring pre-defined transcripts in biological samples to analyze a custom subset of 95 autophagy-related genes in human umbilical vein endothelial cells treated with ultrapure human recombinant endorepellin. We discovered an endorepellin-evoked pro-autophagic and pro-mitophagic gene expression signatures, which included two coordinately up-regulated mitochondrial-associated genes encoding the E3 ubiquitin protein ligase Parkin and the tumor suppressor mitostatin. Induction of both proteins required the tyrosine kinase activity of vascular endothelial growth factor receptor 2 (VEGFR2). Furthermore, we discovered that endorepellin evoked mitochondrial depolarization in endothelial cells via a specific interaction between its two proximal LG1/2 domains and VEGFR2. We also found that following loss of membrane potential, mitostatin and parkin interact and that mitostatin associates with the established Parkin receptor mitofusin-2. In conclusion, we have identified a critical role for endorepellin in remodeling the autophagic transcriptome and influencing mitochondrial homeostasis.


Assuntos
Autofagia , Proteoglicanas de Heparan Sulfato/metabolismo , Células Endoteliais da Veia Umbilical Humana/citologia , Fragmentos de Peptídeos/metabolismo , GTP Fosfo-Hidrolases/genética , GTP Fosfo-Hidrolases/metabolismo , Proteoglicanas de Heparan Sulfato/genética , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Proteínas Mitocondriais/genética , Proteínas Mitocondriais/metabolismo , Fragmentos de Peptídeos/genética , Ligação Proteica , Transdução de Sinais , Transcriptoma , Proteínas Supressoras de Tumor/genética , Proteínas Supressoras de Tumor/metabolismo , Ubiquitina-Proteína Ligases/genética , Ubiquitina-Proteína Ligases/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/genética , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo
14.
BMC Cancer ; 18(1): 687, 2018 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-29940912

RESUMO

BACKGROUND: Heparan sulfate proteoglycans (HSPGs) are complex molecules which play a role in the invasion and growth and metastatic properties of cancerous cells. In this work we analyze changes in the patterns of expression of HSPGs in left sided colorectal cancer (LSCRC), both metastatic and non-metastatic, and the results are also compared with those previously obtained for right sided tumors (RSCRCs). METHODS: Eighteen LSCRCs were studied using qPCR to analyze the expression of both the proteoglycan core proteins and the enzymes involved in heparan sulfate chain biosynthesis. Certain HSPGs also carry chondroitin sulfate chains and so we also studied the genes involved in its biosynthesis. The expression of certain genes that showed significant expression differences were also analysed using immunohistochemical techniques. RESULTS: Changes in proteoglycan core proteins were dependent on their location, and the main differences between metastatic and non-metastatic tumors affected cell-surface glypicans, while other molecules were quite similar. Glypicans were also responsible for the main differences between RS- and LS- malignances. Regarding the biosynthesis of heparan sulfate chains, differential alterations in transcription depending on the presence or not of metastasis affected genes involved in the modification of uronic acid (epimerization and 2-O sulfation), and some isoforms responsible for sulfation of glucosamine (NDST1, HS6ST1). Moreover, in RSCRCs differences were preferentially found in the expression of genes involved in C6 and C3 sulfation of glucosamine, but not in NDSTs or SULFs. Finally, synthesis of chondroitin sulfate showed some alterations, which affected various steps, including polimerization and the modification of chains, but the main variations dependent on the presence of metastases were epimerization and 6C sulfation; however, when compared with RSCRCs, the essential divergences affected polymerization of the chains and the 6C sulfation of the galactosamine residue. CONCLUSIONS: We evidenced alterations in the expression of HSPGs, including the expression of cell surface core proteins, many glycosiltransferases and some enzymes that modify the GAG chains in LSCRCs, but this was dependent on the metastatic nature of the tumor. Some of these alterations are shared with RSCRCs, while others, focused on specific gene groups, are dependent on tumor localization.


Assuntos
Neoplasias Colorretais/patologia , Proteoglicanas de Heparan Sulfato/genética , Idoso , Idoso de 80 Anos ou mais , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/mortalidade , Feminino , Glicosiltransferases/genética , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Metástase Neoplásica , Estadiamento de Neoplasias
15.
Sci Rep ; 8(1): 7262, 2018 05 08.
Artigo em Inglês | MEDLINE | ID: mdl-29740048

RESUMO

Interrupting the interplay between cancer cells and extracellular matrix (ECM) is a strategy to halt tumor progression and stromal invasion. Perlecan/heparan sulfate proteoglycan 2 (HSPG2) is an extracellular proteoglycan that orchestrates tumor angiogenesis, proliferation, differentiation and invasion. Metastatic prostate cancer (PCa) cells degrade perlecan-rich tissue borders to reach bone, including the basement membrane, vasculature, reactive stromal matrix and bone marrow. Domain IV-3, perlecan's last 7 immunoglobulin repeats, mimics native proteoglycan by promoting tumoroid formation. This is reversed by matrilysin/matrix metalloproteinase-7 (MMP-7) cleavage to favor cell dispersion and tumoroid dyscohesion. Both perlecan and Domain IV-3 induced a strong focal adhesion kinase (FAK) dephosphorylation/deactivation. MMP-7 cleavage of perlecan reversed this, with FAK in dispersed tumoroids becoming phosphorylated/activated with metastatic phenotype. We demonstrated Domain IV-3 interacts with the axon guidance protein semaphorin 3A (Sema3A) on PCa cells to deactivate pro-metastatic FAK. Sema3A antibody mimicked the Domain IV-3 clustering activity. Direct binding experiments showed Domain IV-3 binds Sema3A. Knockdown of Sema3A prevented Domain IV-3-induced tumoroid formation and Sema3A was sensitive to MMP-7 proteolysis. The perlecan-Sema3A complex abrogates FAK activity and stabilizes PCa cell interactions. MMP-7 expressing cells destroy the complex to initiate metastasis, destroy perlecan-rich borders, and favor invasion and progression to lethal bone disease.


Assuntos
Proteoglicanas de Heparan Sulfato/genética , Metaloproteinase 7 da Matriz/genética , Neoplasias da Próstata/genética , Semaforina-3A/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Matriz Extracelular/genética , Matriz Extracelular/metabolismo , Proteína-Tirosina Quinases de Adesão Focal/genética , Regulação Neoplásica da Expressão Gênica/genética , Técnicas de Silenciamento de Genes , Humanos , Masculino , Invasividade Neoplásica/genética , Invasividade Neoplásica/patologia , Fosforilação , Próstata/metabolismo , Próstata/patologia , Neoplasias da Próstata/patologia
16.
Sci Rep ; 8(1): 7766, 2018 05 17.
Artigo em Inglês | MEDLINE | ID: mdl-29773865

RESUMO

Perlecan (HSPG2), a heparan sulfate proteoglycan, is a component of basement membranes and participates in a variety of biological activities. Here, we show physiological roles of perlecan in both obesity and the onset of metabolic syndrome. The perinatal lethality-rescued perlecan knockout (Hspg2-/--Tg) mice showed a smaller mass and cell size of white adipose tissues than control (WT-Tg) mice. Abnormal lipid deposition, such as fatty liver, was not detected in the Hspg2-/--Tg mice, and those mice also consumed more fat as an energy source, likely due to their activated fatty acid oxidation. In addition, the Hspg2-/--Tg mice demonstrated increased insulin sensitivity. Molecular analysis revealed the significantly relatively increased amount of the muscle fiber type IIA (X) isoform and a larger quantity of mitochondria in the skeletal muscle of Hspg2-/--Tg mice. Furthermore, the perlecan-deficient skeletal muscle also had elevated levels of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α) protein. PGC1α expression is activated by exercise, and induces mitochondrial biosynthesis. Thus, perlecan may act as a mechano-regulator of catabolism of both lipids and glucose by shifting the muscle fiber composition to oxidative fibers. Our data suggest that downregulation of perlecan is a promising strategy to control metabolic syndrome.


Assuntos
Tecido Adiposo/metabolismo , Metabolismo Energético/fisiologia , Proteoglicanas de Heparan Sulfato/fisiologia , Músculo Esquelético/metabolismo , Animais , Glucose/metabolismo , Proteoglicanas de Heparan Sulfato/genética , Proteoglicanas de Heparan Sulfato/metabolismo , Metabolismo dos Lipídeos , Síndrome Metabólica/metabolismo , Camundongos Knockout , Obesidade/genética , Obesidade/metabolismo , Obesidade/patologia , Condicionamento Físico Animal
17.
Mol Genet Genomic Med ; 6(3): 452-456, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29526034

RESUMO

BACKGROUND: Dyssegmental dysplasia Silverman-Handmaker (DDSH; MIM 224410) type is an extremely rare skeletal dysplasia caused by functional null mutations in the perlecan gene. Less than forty cases are reported in the literature, of which only four were prenatally detected. METHODS: We report on a dizygotic twin pregnancy from consanguineous parents for which one of the twins presented prenatally with severe micromelia, limb bowing and scoliosis, and postnatally with clinical and radiological features compatible with a diagnosis of dyssegmental dysplasia. Molecular studies were undertaken to confirm the clinical diagnosis of DDSH. RESULTS: Molecular analysis results revealed a novel homozygous variant in the HSPG2 gene (MIM 142461), NM_005529.6(HSPG2):c.4029 + 1G>A, consistent with a diagnosis of DDSH. CONCLUSION: To the best of our knowledge, the current report is only the seventh molecularly confirmed case of DDSH.


Assuntos
Nanismo/diagnóstico , Nanismo/genética , Diagnóstico Pré-Natal/métodos , Anormalidades Múltiplas/diagnóstico por imagem , Consanguinidade , Feminino , Desenvolvimento Fetal/genética , Feto/diagnóstico por imagem , Proteoglicanas de Heparan Sulfato/genética , Proteoglicanas de Heparan Sulfato/metabolismo , Humanos , Recém-Nascido , Masculino , Osteocondrodisplasias/genética , Gravidez , Gravidez de Gêmeos , Gêmeos Dizigóticos
18.
Nat Commun ; 9(1): 800, 2018 02 23.
Artigo em Inglês | MEDLINE | ID: mdl-29476074

RESUMO

Human aging is associated with a decline in skeletal muscle (SkM) function and a reduction in the number and activity of satellite cells (SCs), the resident stem cells. To study the connection between SC aging and muscle impairment, we analyze the whole genome of single SC clones of the leg muscle vastus lateralis from healthy individuals of different ages (21-78 years). We find an accumulation rate of 13 somatic mutations per genome per year, consistent with proliferation of SCs in the healthy adult muscle. SkM-expressed genes are protected from mutations, but aging results in an increase in mutations in exons and promoters, targeting genes involved in SC activity and muscle function. In agreement with SC mutations affecting the whole tissue, we detect a missense mutation in a SC propagating to the muscle. Our results suggest somatic mutagenesis in SCs as a driving force in the age-related decline of SkM function.


Assuntos
Envelhecimento/genética , Músculo Esquelético/crescimento & desenvolvimento , Mutação , Células Satélites de Músculo Esquelético/citologia , Adulto , Idoso , Envelhecimento/metabolismo , Diferenciação Celular , Proliferação de Células , Conectina/genética , Conectina/metabolismo , Citocinas/genética , Citocinas/metabolismo , Éxons , Feminino , Fibronectinas , Proteoglicanas de Heparan Sulfato/genética , Proteoglicanas de Heparan Sulfato/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Músculo Esquelético/citologia , Músculo Esquelético/metabolismo , Mutagênese , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Regiões Promotoras Genéticas , Células Satélites de Músculo Esquelético/metabolismo , Adulto Jovem
19.
Viruses ; 10(1)2018 01 03.
Artigo em Inglês | MEDLINE | ID: mdl-29301346

RESUMO

It has been proposed that blood coagulation factors, principally factor X (FX), enhance the uptake of human adenovirus type 5 (Ad5) into cultured epithelial cells by bridging the viral hexon capsid protein and cell-surface heparan sulphate proteoglycans (HSPGs). We studied the effects of FX on Ad transduction of lymphoid cell lines (NK92MI, a natural killer cell line; Daudi, a B-cell line and Jurkat, a T-cell line) as well as primary peripheral blood lymphocytes (PBL) and HeLa epithelial cells using either replication-deficient Ad5, or a derivative in which the Ad5 fiber was replaced with that of another Ad type, Ad35, termed Ad5F35. PBL and NK92MI were resistant to Ad5 transduction. Transduction of Jurkat and Daudi cells by Ad5 was reduced by FX but without discernible effects on cell-surface Ad5 binding. FX reduced virus binding and transduction of all lymphoid cell lines by Ad5F35, as well as transduction of the T- and Natural Killer (NK)-cell populations of PBL. Flow cytometry analysis showed that all lymphoid cell lines were negative for HSPG components, in contrast to HeLa cells. FX reduced transduction of an HSPG-negative mutant Chinese hamster ovary cell line (CHOpgsA745) by Ad5 and Ad5F35, with Ad5F35 binding also being reduced by FX. These results point to fiber-dependent differences (Ad5 versus Ad35 fiber) in Ad binding to and transduction of human lymphoid and epithelial cells in the presence of FX.


Assuntos
Infecções por Adenovirus Humanos/metabolismo , Infecções por Adenovirus Humanos/virologia , Adenovírus Humanos/fisiologia , Fator X/metabolismo , Linfócitos/metabolismo , Linfócitos/virologia , Internalização do Vírus , Infecções por Adenovirus Humanos/imunologia , Animais , Células CHO , Cricetulus , Células Epiteliais/metabolismo , Células Epiteliais/virologia , Células HeLa , Proteoglicanas de Heparan Sulfato/genética , Proteoglicanas de Heparan Sulfato/metabolismo , Humanos , Transdução Genética
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA