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1.
PLoS One ; 15(5): e0227527, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32374776

RESUMO

Type 2 diabetes and obesity are associated with back pain in juveniles and adults and are implicated in intervertebral disc (IVD) degeneration. Hypercaloric Western diets are associated with both obesity and type 2 diabetes. The objective of this study was to determine if obesity and type 2 diabetes result in spinal pathology in a sex-specific manner using in vivo diabetic and dietary mouse models. Leptin is an appetite-regulating hormone, and its deficiency leads to polyphagia, resulting in obesity and diabetes. Leptin is also associated with IVD degeneration, and increased expression of its receptor was identified in degenerated IVDs. We used young, leptin receptor deficient (Db/Db) mice to mimic the effect of diet and diabetes on adolescents. Db/Db and Control mice were fed either Western or Control diets, and were sacrificed at 3 months of age. Db/Db mice were obese, while only female mice developed diabetes. Female Db/Db mice displayed altered IVD morphology, with increased intradiscal notochordal band area, suggesting delayed IVD cell proliferation and differentiation, rather than IVD degeneration. Motion segments from Db/Db mice exhibited increased failure risk with decreased torsional failure strength. Db/Db mice also had inferior bone quality, which was most prominent in females. We conclude that obesity and diabetes due to impaired leptin signaling contribute to pathological changes in vertebrae, as well as an immature IVD phenotype, particularly of females, suggesting a sex-dependent role of leptin in the spine.


Assuntos
Diabetes Mellitus Tipo 2/genética , Degeneração do Disco Intervertebral/genética , Leptina/genética , Obesidade/genética , Receptores para Leptina/genética , Animais , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/patologia , Dieta Ocidental/efeitos adversos , Modelos Animais de Doenças , Feminino , Humanos , Disco Intervertebral/metabolismo , Disco Intervertebral/patologia , Degeneração do Disco Intervertebral/metabolismo , Degeneração do Disco Intervertebral/patologia , Leptina/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos NOD , Obesidade/metabolismo , Obesidade/patologia , Receptores para Leptina/deficiência , Caracteres Sexuais , Transdução de Sinais/genética , Coluna Vertebral/metabolismo , Coluna Vertebral/patologia
2.
Gene ; 750: 144634, 2020 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-32240779

RESUMO

Lumbar degenerative disc disease (DDD) is a multifaceted progressive condition and often accompanied by disc herniation (DH) and/or degenerative spondylolisthesis (DS). Given the high prevalence of the disease (up to 20% according to some estimates) and the high costs associated with its care, there is a need to explore novel therapies such as regenerative medicine. Exploring these novel therapies first warrants investigation of molecular pathways underlying these disorders. Here, we show results from next generation RNA sequencing (RNA-seq) on mRNA isolated from 10 human nucleus pulposus (NP) samples of lumbar degenerated discs (DH and DS; n = 5 for each tissue) and other musculoskeletal tissues (Bone, cartilage, growth plate, and muscle; n = 7 for each tissue). Pathway and network analyses based on gene ontology (GO) terms were used to identify the biological functions of differentially expressed mRNAs. A total of 701 genes were found to be significantly upregulated in lumbar NP tissue compared to other musculoskeletal tissues. These differentially expressed mRNAs were primarily involved in DNA damage, immunity and G1/S transition of mitotic cell cycle. Interestingly, DH-specific signaling genes showed major network in chemotactic (e.g., CXCL10, CXCL11, IL1RL2 and IL6) and matrix-degrading pathway (e.g., MMP16, ADAMTSL1, 5, 8, 12, and 15), while DS-specific signaling genes were found to be those involved in cell adhesion (e.g., CDH1, EPHA1 and EFNA2) and inflammatory cytokines (e.g., CD19, CXCL5, CCL24, 25 and XCL2). Our findings provide new leads for therapeutic drug discovery that would permit optimization of medical or pharmacological intervention for cases of lumbar DDD.


Assuntos
Degeneração do Disco Intervertebral/genética , Deslocamento do Disco Intervertebral/genética , Espondilolistese/genética , Adulto , Cartilagem/metabolismo , Citocinas/metabolismo , Feminino , Ontologia Genética , Redes Reguladoras de Genes/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Humanos , Disco Intervertebral/metabolismo , Vértebras Lombares/metabolismo , Masculino , Núcleo Pulposo/metabolismo , RNA Mensageiro/metabolismo
3.
Br J Neurosurg ; 34(1): 66-71, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32141366

RESUMO

Background: Intervertebral disc degeneration (IVDD) is a multifactorial disease that is sensitive to the balance between anti-inflammatory and pro-inflammatory cytokines. This study investigated the single nucleotide polymorphisms (SNPs) of interleukin 4 (IL-4) in IVDD.Methods: Genomic DNA of peripheral mononuclear cells of 76 IVDD patients and 140 healthy controls were investigated for three SNPs of IL-4 (rs2243248 (-1098G/T), rs2243250 (-590 C/T), rs2070874 (-33 C/T)) and 1 SNP of IL-4RA (rs180275, +1902 A/G) through PCR-SSP method.Results: The 'C' allele frequency of IL-4 rs2243250 was 104 in 76 patients, while it was 149 in 140 controls (OR = 2, p = .001); also this SNP was significantly associated with post-operative pain reduction. The 'C' allele of IL-4 rs2070874 (130 in 76 patients, and 200 in 140 controls, OR = 2.66), and the 'CC' genotype were more frequent among patients (OR = 3.98, p < .001) than controls. 'TTT' haplotype was more common in controls (OR = 0.36, p < .001) and 'TCC' was also more common in patients (OR = 1.75, p = .012). A meta-analysis of previous studies found significantly higher IL-4 levels in disc tissues of IVDD patients, which was not similarly found in blood samples.Conclusion: The immune system plays an important role in IVDD. The extent and progress of the disease vary significantly with IL-4 level. Meanwhile, the rs2070874 and rs2243250 SNPs of IL-4 were significantly associated with IVDD in Iranian patients.


Assuntos
Subunidade alfa de Receptor de Interleucina-4/genética , Interleucina-4/genética , Degeneração do Disco Intervertebral/genética , Adulto , Estudos de Casos e Controles , Feminino , Frequência do Gene , Genótipo , Humanos , Interleucina-4/biossíntese , Subunidade alfa de Receptor de Interleucina-4/biossíntese , Degeneração do Disco Intervertebral/epidemiologia , Irã (Geográfico)/epidemiologia , Masculino , Pessoa de Meia-Idade , Dor Pós-Operatória/epidemiologia , Dor Pós-Operatória/genética , Polimorfismo de Nucleotídeo Único , Adulto Jovem
4.
Mol Med Rep ; 21(3): 1163-1171, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31922222

RESUMO

Approximately 50% of the cases of low back pain (LBP) are attributed to discogenic origin. The causes of discogenic pain are complicated and consist of a complex biochemical cascade. Neovascularization of intervertebral discs (IVDs) is believed to be associated with discogenic pain. The anti­angiogenesis ability of tissue inhibitor of metalloproteinase­3 (TIMP3) has been reported in many tumors, yet whether TIMP3 is associated with neovascularization of IVDs remains unknown. In the present study, both in vitro and in vivo models were used to investigate the association between discogenic pain and TIMP3 expression in nucleus pulposus (NP). PCR results demonstrated that inflammation induced downregulation of TIMP3 expression in NP cells. By using an adenovirus system to upregulate TIMP3 expression, the effect of TIMP3 on angiogenesis was measured by endothelial cell migration and tube formation assays. The results demonstrated that overexpression of TIMP3 suppressed angiogenesis in NP without the regulation of vascular endothelial growth factor (VEGF) expression. TNF­α converting enzyme (TACE) expression was downregulated by TIMP3, thus inhibiting the TACE­induced activation of TNF­α in NP cells. Immunohistochemical staining of IVDs also confirmed that TIMP3 inhibited the expression of substance P in NP. Taken together, the present results indicated the expression of TIMP3 in NP may have a key role in the development of discogenic pain.


Assuntos
Dor nas Costas/metabolismo , Degeneração do Disco Intervertebral/metabolismo , Neovascularização Patológica/metabolismo , Núcleo Pulposo , Substância P/biossíntese , Inibidor Tecidual de Metaloproteinase-3/antagonistas & inibidores , Regulação para Cima , Adenoviridae , Animais , Dor nas Costas/genética , Dor nas Costas/patologia , Vetores Genéticos , Degeneração do Disco Intervertebral/genética , Degeneração do Disco Intervertebral/patologia , Degeneração do Disco Intervertebral/terapia , Masculino , Neovascularização Patológica/genética , Neovascularização Patológica/patologia , Neovascularização Patológica/terapia , Núcleo Pulposo/irrigação sanguínea , Núcleo Pulposo/metabolismo , Núcleo Pulposo/fisiologia , Ratos , Ratos Sprague-Dawley , Substância P/genética , Inibidor Tecidual de Metaloproteinase-3/genética , Transdução Genética
5.
Spine (Phila Pa 1976) ; 45(11): E616-E623, 2020 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-31923126

RESUMO

STUDY DESIGN: Meta-analysis to collect relevant studies to assess the association between COL11A1 and GDF5 genetic variants and susceptibility to IDD. OBJECTIVE: The aim of this study was to assess whether or not COL11A1 and GDF5 genetic variants were associated with susceptibility to IDD. SUMMARY OF BACKGROUND DATA: IDD or LDH is a major public health problem. There have been several studies evaluating the relationship between COL11A1 and GDF5 genetic variants with risk of intervertebral disc degeneration (IDD). However, the studies were limited in discrete outcome and sample size, and some of the results were contradictory. METHODS: We systematically searched the relevant publications in electronic databases. Eligible studies were included based on the defined criteria. The pooled odds ratios (ORs) with its 95% confidence intervals (CIs) were received using STATA 15. Subgroup analysis, sensitivity analysis, publication bias, and the "Trim and fill" method were performed in the meta-analysis. RESULTS: A total of 3287 IDD cases and 5115 controls were incorporated into the meta-analysis. Our results demonstrated that COL11A1 rs1676486 was significantly associated with increased IDD susceptibility under all genetic models (allele model T vs. C: OR = 1.40, 95% CI 1.23-1.59, P = 0.000; homozygote model TT vs. CC: OR = 1.89, 95%CI 1.40-2.56, P = 0.000; dominant model TT+TC vs. CC: OR = 1.52, 95% CI 1.29-1.80, P = 0.000; recessive model TT vs. TC + CC: OR = 1.58, 95% CI 1.18-2.12, P = 0.002). However, GDF5 rs143383 was not (allele model T vs. C: OR = 1.15, 95% CI 0.91-1.44, P = 0.244; homozygote model TT vs. CC: OR = 1.22, 95% CI 0.75-2.00, P = 0.429; dominant model TT vs. CC+CT: OR = 1.22, 95% CI 0.95-1.57, P = 0.112; recessive model TC + TT vs. CC: OR = 1.12, 95% CI 0.73-1.73, P = 0.594). Subgroup analysis indicated ethnicity was not the source of heterogeneity. Sensitivity analysis, publication bias, and the "Trim and fill" method demonstrated the meta-analysis was of reliability. CONCLUSION: Our results suggested that COL11A1 rs1676486 was significantly associated with IDD and the T allele was a risky factor. However, GDF5 rs143383 was not. LEVEL OF EVIDENCE: 1.


Assuntos
Colágeno Tipo XI/genética , Predisposição Genética para Doença/genética , Variação Genética/genética , Fator 5 de Diferenciação de Crescimento/genética , Degeneração do Disco Intervertebral/genética , Humanos , Degeneração do Disco Intervertebral/diagnóstico , Polimorfismo de Nucleotídeo Único/genética , Reprodutibilidade dos Testes , Fatores de Risco
6.
FASEB J ; 34(2): 1970-1982, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31909538

RESUMO

Osterix is a critical transcription factor of mesenchymal stem cell fate, where its loss or loss of Wnt signaling diverts differentiation to a chondrocytic lineage. Intervertebral disc (IVD) degeneration activates the differentiation of prehypertrophic chondrocyte-like cells and inactivates Wnt signaling, but its interactive role with osterix is unclear. First, compared to young-adult (5 mo), mechanical compression of old (18 mo) IVD induced greater IVD degeneration. Aging (5 vs 12 mo) and/or compression reduced the transcription of osterix and notochordal marker T by 40-75%. Compression elevated the transcription of hypertrophic chondrocyte marker MMP13 and pre-osterix transcription factor RUNX2, but less so in 12 mo IVD. Next, using an Ai9/td reporter and immunohistochemical staining, annulus fibrosus and nucleus pulposus cells of young-adult IVD expressed osterix, but aging and compression reduced its expression. Lastly, in vivo LRP5-deficiency in osterix-expressing cells inactivated Wnt signaling in the nucleus pulposus by 95%, degenerated the IVD to levels similar to aging and compression, reduced the biomechanical properties by 45-70%, and reduced the transcription of osterix, notochordal markers and chondrocytic markers by 60-80%. Overall, these data indicate that age-related inactivation of Wnt signaling in osterix-expressing cells may limit regeneration by depleting the progenitors and attenuating the expansion of chondrocyte-like cells.


Assuntos
Envelhecimento/metabolismo , Condrócitos/metabolismo , Condrogênese , Subunidade alfa 1 de Fator de Ligação ao Core/biossíntese , Proteínas Fetais/biossíntese , Degeneração do Disco Intervertebral/metabolismo , Fator de Transcrição Sp7/biossíntese , Proteínas com Domínio T/biossíntese , Envelhecimento/genética , Envelhecimento/patologia , Animais , Antígenos de Diferenciação/biossíntese , Antígenos de Diferenciação/genética , Condrócitos/patologia , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Proteínas Fetais/genética , Regulação da Expressão Gênica , Degeneração do Disco Intervertebral/genética , Degeneração do Disco Intervertebral/patologia , Camundongos , Camundongos Transgênicos , Fator de Transcrição Sp7/genética , Proteínas com Domínio T/genética
7.
PLoS One ; 14(11): e0225527, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31751427

RESUMO

Rabbits with naturally high levels of cholesterol ester transfer protein (CETP), unlike rodents, have become an interesting animal model for the study of lipid metabolism and atherosclerosis, as they have similarities to humans in lipid metabolism, cardiovascular physiology and susceptibility to develop atherosclerosis. Rodents, such as mice, are not prone to atherosclerosis as they lack the mass and activity of CETP, as a key player in lipoprotein metabolism. Recently, APOE-knockout in rabbits has been shown to promote atherosclerosis and associated premature IVD degeneration that mimic the symptoms of atherosclerosis and structural changes of IVDs in humans. Here we examined whether APOE-knockout promoted IVD degeneration in rabbits is associated with imbalanced inflammatory catabolic activities, as the underlying problem of biological deterioration that mimic the symptoms of advanced IVD degeneration in humans. We analysed in lumbar nucleus pulposus (NP) of APOE-knockout rabbits the cell viabilities and the intracellular levels of inflammatory, catabolic, anti-catabolic and anabolic proteins derogating IVD matrix. Grades of IVD degeneration were evaluated by magnetic resonance imaging. NP cells were isolated from homozygous APOE-knockout and wild-type New Zealand White rabbits of similar age. Three-dimensional cell culture with low-glucose was completed in alginate hydrogel. Cell proliferation and intracellular levels of target proteins were examined by MTT and ELISA assays. Alike human NP cells of different disc degeneration grades, NP cells of APOE-knockout and wild-type rabbits showed significantly different in vivo cell population densities (p<0.0001) and similar in vitro proliferation rates. Furthermore, they showed differences in overexpression of selective inflammatory and catabolic proteins (p<0.0001) similar to those found in human NP cells of different disc degeneration grades, such as IL-1ß, TNF-α, ADAMTS-4, ADAMTS-5 and MMP-3. This study showed that premature IVD degeneration in APOE-knockout rabbits was promoted by the accumulation of selective inflammatory catabolic factors that enhanced imbalances between catabolic and anabolic factors mimicking the symptoms of advanced IVD degeneration in humans. Thus, APOE-knockout rabbits could be used as a promising model for therapeutic approaches of degenerative disc disorders.


Assuntos
Apolipoproteínas E/genética , Citocinas/metabolismo , Degeneração do Disco Intervertebral/diagnóstico por imagem , Núcleo Pulposo/citologia , Animais , Técnicas de Cultura de Células , Proliferação de Células , Células Cultivadas , Proteínas de Transferência de Ésteres de Colesterol , Modelos Animais de Doenças , Regulação da Expressão Gênica , Técnicas de Inativação de Genes , Degeneração do Disco Intervertebral/genética , Degeneração do Disco Intervertebral/imunologia , Imagem por Ressonância Magnética , Masculino , Núcleo Pulposo/imunologia , Coelhos
8.
Mol Med Rep ; 20(6): 4875-4882, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31638197

RESUMO

Intervertebral disc degeneration (IDD) is characterized by abnormal induction of apoptosis in intervertebral disc nucleus pulposus (NP) cells. Previous studies indicated that miR­222 was upregulated in patients with rheumatoid arthritis. However, the effects of miR­222 in IDD remain unclear. The present study aimed to demonstrate the role of miR­222 in NP cells. The levels of miR­222 in patients with IDD were measured by reverse transcription­quantitative PCR. Cell Counting Kit­8 and western blotting assays were used to detect cell proliferation and apoptosis­associated protein levels, respectively. In addition, luciferase reporter assays were performed to validate the predicted target genes of miR­222. miR­222 was significantly upregulated in patients with IDD. Overexpression of miR­222 inhibited cell proliferation and induced cell apoptosis. Moreover, overexpression of miR­222 resulted in an upregulation in the levels of Bax and cleaved caspase 3, and a downregulation in the levels of Bcl­2 in NP cells. The luciferase reporter assays demonstrated that Bcl­2 is a target of miR­222. Furthermore, overexpression of miR­222 increased the levels of cytochrome c, apoptotic protease activating factor­1 and cleaved caspase 9 in NP cells. Conversely, downregulation of miR­222 could promote the proliferation of NP cells. The present data demonstrated that miR­222 induced apoptosis in NP cells by directly targeting Bcl­2. Therefore, miR­222 may act as a potential therapeutic target for the treatment of IDD.


Assuntos
Apoptose , Degeneração do Disco Intervertebral/genética , MicroRNAs/genética , Núcleo Pulposo/citologia , Proteínas Proto-Oncogênicas c-bcl-2/genética , Adulto , Idoso , Linhagem Celular , Regulação para Baixo , Feminino , Humanos , Disco Intervertebral/citologia , Disco Intervertebral/metabolismo , Masculino , Pessoa de Meia-Idade , Núcleo Pulposo/metabolismo , Regulação para Cima
9.
PLoS Genet ; 15(10): e1008096, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31652254

RESUMO

Degenerative changes of the intervertebral disc (IVD) are a leading cause of disability affecting humans worldwide and has been attributed primarily to trauma and the accumulation of pathology during aging. While genetic defects have also been associated with disc degeneration, the precise mechanisms driving the initiation and progression of disease have remained elusive due to a paucity of genetic animal models. Here, we discuss a novel conditional mouse genetic model of endplate-oriented disc herniations in adult mice. Using conditional mouse genetics, we show increased mechanical stiffness and reveal dysregulation of typical gene expression profiles of the IVD in adhesion G-protein coupled receptor G6 (Adgrg6) mutant mice prior to the onset of endplate-oriented disc herniations in adult mice. We observed increased STAT3 activation prior to IVD defects and go on to demonstrate that treatment of Adgrg6 conditional mutant mice with a small molecule inhibitor of STAT3 activation ameliorates endplate-oriented herniations. These findings establish ADGRG6 and STAT3 as novel regulators of IVD endplate and growth plate integrity in the mouse, and implicate ADGRG6/STAT3 signaling as promising therapeutic targets for endplate-oriented disc degeneration.


Assuntos
Degeneração do Disco Intervertebral/genética , Deslocamento do Disco Intervertebral/genética , Receptores Acoplados a Proteínas-G/genética , Fator de Transcrição STAT3/genética , Animais , Modelos Animais de Doenças , Progressão da Doença , Lâmina de Crescimento , Humanos , Disco Intervertebral/crescimento & desenvolvimento , Disco Intervertebral/patologia , Degeneração do Disco Intervertebral/fisiopatologia , Deslocamento do Disco Intervertebral/fisiopatologia , Camundongos , Mutação , Transdução de Sinais
10.
Oxid Med Cell Longev ; 2019: 7959573, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31583043

RESUMO

Intervertebral disc degeneration (IDD) is one of the major causes of low back pain. Diabetes is a risk factor for IDD and may aggravate IDD in rats; however, the mechanism is poorly understood. Previously, we demonstrated that apoptosis and senescence were increased in diabetic nucleus pulposus (NP) tissues; in the current study, we found that hyperglycaemia may promote the incidence of apoptosis and senescence in NP cells in vitro. Meanwhile, the acetylation of P53, a master transcription factor of apoptosis and senescence, was also found increased in diabetic NP tissues in vivo as well as in hyperglycaemic NP cells in vitro. Sirt1 is an NAD+-dependent deacetylase, and we showed that the expression of Sirt1 was decreased in NP tissues, while hyperglycaemia could suppress the expression and activity of Sirt1 in NP cells. Furthermore, we demonstrated that butein may inhibit acetylation of P53 and protect NP cells against hyperglycaemia-induced apoptosis and senescence through Sirt1 activation, as the Sirt1 inhibitor Ex527 may counteract the protective effect of butein in hyperglycaemic NP cells. An in vivo study showed that butein could ameliorate the IDD process in diabetic rats, while Sirt1 was increased and acetyl-p53 was decreased in NP tissues in butein-treated rats. These results indicate that the Sirt1/P53 axis is involved in the pathogenesis of diabetic IDD and may serve as a therapeutic target for diabetic IDD.


Assuntos
Diabetes Mellitus Experimental/complicações , Degeneração do Disco Intervertebral/genética , Sirtuína 1/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Animais , Modelos Animais de Doenças , Degeneração do Disco Intervertebral/patologia , Masculino , Ratos , Ratos Sprague-Dawley
11.
Vet Pathol ; 56(6): 895-902, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31526126

RESUMO

Chondrodystrophy results in predictable and progressive biochemical and structural changes to the intervertebral disc, resulting in early onset degeneration and dystrophic mineralization of the disc. Accelerated degeneration and mineralization of the intervertebral disc are common in multiple dog breeds and can result in compromised function, herniation, pain, and a variety of neurological sequelae. A mutation responsible for chondrodystrophy in dogs has been identified as an aberrant fibroblast growth factor 4 (FGF4) retrogene insertion on chromosome 12 (CFA12) and is associated with short stature of the Nova Scotia Duck Tolling Retriever. Segregation of the CFA12 FGF4 retrogene in this dog breed provides an opportunity to examine the effect of retrogene presence on radiographic and histologic appearance of chondrodystrophic disc degeneration within a single breed. Here we found that in the intervertebral discs isolated from 2 dogs with the CFA12 FGF4 genotype, the nucleus pulposus was largely replaced by cartilaginous tissue, and physaliferous notochordal cells were rarely if ever identified. These findings are in contrast to the normal histologic findings in 2 breed-matched dogs lacking the mutation. The findings are consistent with premature chondroid degeneration of the intervertebral disc and suggest that the presence of the CFA12 FGF4 retrogene is sufficient to cause the chondrodystrophic phenotype.


Assuntos
Doenças das Cartilagens/veterinária , Doenças do Cão/patologia , Fator 4 de Crescimento de Fibroblastos/genética , Degeneração do Disco Intervertebral/veterinária , Animais , Doenças das Cartilagens/diagnóstico , Doenças das Cartilagens/genética , Doenças das Cartilagens/patologia , Doenças do Cão/diagnóstico , Doenças do Cão/genética , Cães , Genótipo , Disco Intervertebral/patologia , Degeneração do Disco Intervertebral/diagnóstico , Degeneração do Disco Intervertebral/genética , Degeneração do Disco Intervertebral/patologia , Fenótipo
12.
PLoS One ; 14(9): e0222188, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31513634

RESUMO

BACKGROUND: Environmental and endogenous factors under genetic predisposition are considered to initiate the human intervertebral disc (IVD) degeneration. DNA methylation is an essential mechanism to ensure cell-specific gene expression for normal development and tissue stability. Aberrant epigenetic alterations play a pivotal role in several diseases, including osteoarthritis. However, epigenetic alternations, including DNA methylation, in IVD degeneration have not been evaluated. The purpose of this study was to comprehensively compare the genome-wide DNA methylation profiles of human IVD tissues, specifically nucleus pulpous (NP) tissues, with early and advanced stages of disc degeneration. METHODS: Human NP tissues were used in this study. The samples were divided into two groups: early stage degeneration (n = 8, Pfirrmann's MRI grade: I-III) and advanced stage degeneration (n = 8, grade: IV). Genomic DNA was processed for genome-wide DNA methylation profiling using the Infinium MethylationEPIC BeadChip array. Extraction of raw methylation data, clustering and scatter plot of each group values of each sample were performed using a methylation module in GenomeStudio software. The identification of differentially methylated loci (DMLs) and the Gene Ontology (GO) analysis were performed using R software with the ChAMP package. RESULTS: Unsupervised hierarchical clustering revealed that early and advanced stage degenerated IVD samples segregated into two main clusters by their DNA methylome. A total of 220 DMLs were identified between early and advanced disc degeneration stages. Among these, four loci were hypomethylated and 216 loci were hypermethylated in the advanced disc degeneration stage. The GO enrichment analysis of genes containing DMLs identified two significant GO terms for biological processes, hemophilic cell adhesion and cell-cell adhesion. CONCLUSIONS: We conducted a genome-wide DNA methylation profile comparative study and observed significant differences in DNA methylation profiles between early and advanced stages of human IVD degeneration. These results implicate DNA methylation in the process of human IVD degeneration.


Assuntos
Epigênese Genética/genética , Degeneração do Disco Intervertebral/genética , Núcleo Pulposo/fisiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Ilhas de CpG/genética , Metilação de DNA/genética , Epigenômica/métodos , Feminino , Expressão Gênica/genética , Perfilação da Expressão Gênica/métodos , Genoma/genética , Estudo de Associação Genômica Ampla/métodos , Humanos , Disco Intervertebral/metabolismo , Masculino , Pessoa de Meia-Idade , Núcleo Pulposo/metabolismo
13.
Genet Test Mol Biomarkers ; 23(9): 610-617, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31368816

RESUMO

Aims: This study was designed to investigate differentially expressed genes (DEGs) in the annulus fibrosus (AF), nucleus pulposus (NP), and whole blood (WB) of intervertebral disk degeneration (IDD) patients. Materials and Methods: We retrieved microarray data set GSE70362, which contains the gene expression profiles of 24 AF and 24 NP samples from the Gene Expression Omnibus and identified DEGs in degenerative AF (AF-DEGs) and NP (NP-DEGs) samples compared with nondegenerative samples. We also examined gene expression profiles in WB from patients with IDD and healthy volunteers to identify DEGs in WB (WB-DEGs). We performed functional analyses on the DEGs common to AF-DEGs, NP-DEGs, and WB-DEGs. Expression of the common DEGs was partially validated by quantitative real-time-polymerase chain reaction (QRT-PCR). Results: In total, 846 AF-DEGs, 902 NP-DEGs, and 862 WB-DEGs were identified, and 22 DEGs were common among the three groups. Functional analyses showed that the common DEGs were enriched in 33 biological processes, 16 cellular components, 4 molecular functions, and 9 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways; 13 of the common DEGs were included in the protein-protein interaction (PPI) network and superoxide dismutase 2 (SOD2) was identified as a hub gene in the PPI network. The QRT-PCR results for the expression of the genes protein disulfide isomerase family A member 4, FKBP prolyl isomerase 11, ectonucleotide pyrophosphatase/phosphodiesterase 4, SOD2, and actin binding LIM protein 1, were consistent with the gene chip hybridization results. Conclusions: This study identified key genes for future investigations of the underlying molecular mechanisms of IDD. These genes may provide future targets for the clinical treatment and diagnosis of IDD.


Assuntos
Degeneração do Disco Intervertebral/genética , Transcriptoma , Adulto , Anel Fibroso/metabolismo , Biologia Computacional , Feminino , Perfilação da Expressão Gênica , Humanos , Degeneração do Disco Intervertebral/metabolismo , Proteínas com Domínio LIM/sangue , Proteínas com Domínio LIM/genética , Proteínas com Domínio LIM/metabolismo , Masculino , Análise em Microsséries , Proteínas dos Microfilamentos/sangue , Proteínas dos Microfilamentos/genética , Proteínas dos Microfilamentos/metabolismo , Pessoa de Meia-Idade , Núcleo Pulposo/metabolismo , Diester Fosfórico Hidrolases/sangue , Isomerases de Dissulfetos de Proteínas/sangue , Isomerases de Dissulfetos de Proteínas/genética , Isomerases de Dissulfetos de Proteínas/metabolismo , Mapas de Interação de Proteínas , Superóxido Dismutase/sangue , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo
14.
Spine (Phila Pa 1976) ; 44(23): 1623-1629, 2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31365516

RESUMO

STUDY DESIGN: Genetic case-control study of single nucleotide polymorphisms (SNPs). OBJECTIVE: To examine the association of previously reported susceptibility genes for adolescent idiopathic scoliosis (AIS) and intervertebral disc (IVD) degeneration with adult spinal deformity (ASD). SUMMARY OF BACKGROUND DATA: ASD is a spinal deformity that develops and progresses with age. Its etiology is unclear. Several ASD susceptibility genes were recently reported using a candidate gene approach; however, the sample sizes were small and associations with ASD development were not determined. METHODS: ASD was defined as structural scoliosis with a Cobb angle more than 15° on standing radiographs, taken of patients at age 40 to 75 years in this study. Subjects in whom scoliosis was diagnosed before age 20 were excluded. We recruited 356 Japanese ASD subjects and 3341 healthy controls for case-control association studies of previously reported SNPs. We genotyped four known AIS-associated SNPs (rs11190870 in LBX1, rs6570507 in GPR126, rs10738445 in BNC2, and rs6137473 in PAX1) and three IVD degeneration-associated SNPs (rs1245582 in CHST3, rs2073711 in CILP, and rs1676486 in COL11A1) by the Invader assay. RESULTS: Among the AIS-associated SNPs, rs11190870 and rs6137473 showed strong and nominal associations with ASD (P = 1.44 × 10, 1.00 × 10, respectively). Of the IVD degeneration-associated SNPs, rs1245582 and rs2073711 showed no association with ASD, while rs1676486 showed a nominal association (P = 1.10 × 10). In a subgroup analysis, rs11190870 was significantly associated with a Cobb angle more than 20° in the minor thoracic curve (P = 1.44 × 10) and with a left convex lumbar curve (P = 6.70 × 10), and nominally associated with an apical vertebra higher than L1 (P = 1.80 × 10). CONCLUSION: rs11190870 in LBX1, a strong susceptibility SNP for AIS, may also be a susceptibility SNP for ASD. Thus, ASD and AIS may share a common genetic background. LEVEL OF EVIDENCE: 4.


Assuntos
Estudos de Associação Genética/métodos , Predisposição Genética para Doença/genética , Proteínas de Homeodomínio/genética , Degeneração do Disco Intervertebral/genética , Polimorfismo de Nucleotídeo Único/genética , Escoliose/genética , Fatores de Transcrição/genética , Adulto , Idoso , Estudos de Casos e Controles , Feminino , Predisposição Genética para Doença/epidemiologia , Genótipo , Humanos , Degeneração do Disco Intervertebral/diagnóstico por imagem , Degeneração do Disco Intervertebral/epidemiologia , Masculino , Pessoa de Meia-Idade , Escoliose/diagnóstico por imagem , Escoliose/epidemiologia
15.
Gene ; 715: 144029, 2019 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-31376409

RESUMO

Intervertebral disc degeneration (IDD) is a major cause of lower back pain, but the specific molecular mechanisms governing its development are poorly characterized. This study sought to assess to what extent HOTAIR, a long non-coding (Lnc) RNA is expressed in IDD and regulates the apoptotic death of nucleus pulposus (NP) cells. We therefore used real-time qPCR to measure HOTAIR and microRNA(miR)-34a-5p in degenerative NP cells, and then validated their functional relevance via overexpressing them in these NP cells. We further verified the targets of these RNA constructs in 293 T cells through the use of a dual luciferase reporter assay. We further measured NP cell apoptosis via flow cytometry and Notch1 expression via western blotting. Our results indicated that IDD was linked with decreased HOTAIR expression relative to regular NP cells, and overexpressing this lncRNA was linked to reduced apoptotic NP cell death, whereas overexpressing miR-34a-5p had the opposite effect. We found that HOTAIR served as a miR-34a-5p sponge, sequestering this miRNA and thereby down regulating genes linked to apoptosis through the Notch signaling pathway. Even in naturally degenerated NP cells, HOTAIR delayed the onset of apoptosis. Together these results reveal that a HOTAIR/miR-34a-5p/Notch1 signaling pathway may regulate the development of IDD, potentially making HOTAIR a viable target for treatment of this disease.


Assuntos
Apoptose , Regulação para Baixo , Degeneração do Disco Intervertebral/metabolismo , MicroRNAs/biossíntese , RNA Longo não Codificante/biossíntese , Receptor Notch1/biossíntese , Transdução de Sinais , Adulto , Idoso , Feminino , Humanos , Degeneração do Disco Intervertebral/genética , Degeneração do Disco Intervertebral/patologia , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Núcleo Pulposo/patologia , RNA Longo não Codificante/genética , Receptor Notch1/genética
16.
Int J Mol Sci ; 20(15)2019 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-31344903

RESUMO

Chronic back pain is a common disability, which is often accredited to intervertebral disc degeneration. Gold standard interventions such as spinal fusion, which are mainly designed to mechanically seal the defect, frequently fail to restore the native biomechanics. Moreover, artificial implants have limited success as a repair strategy, as they do not alter the underlying disease and fail to promote tissue integration and subsequent native biomechanics. The reported high rates of spinal fusion and artificial disc implant failure have pushed intervertebral disc degeneration research in recent years towards repair strategies. Intervertebral disc repair utilizing principles of tissue engineering should theoretically be successful, overcoming the inadequacies of artificial implants. For instance, advances in the development of scaffolds aided with cells and growth factors have opened up new possibilities for repair strategies. However, none has reached the stage of clinical trials in humans. In this review, we describe the hitches encountered in the musculoskeletal field and summarize recent advances in designing tissue-engineered constructs for promoting nucleus pulposus repair. Additionally, the review focuses on the effect of biomaterial aided with cells and growth factors on achieving effective functional reparative potency, highlighting the ways to enhance the efficacy of these treatments.


Assuntos
Dor nas Costas/genética , Núcleo Celular/genética , Degeneração do Disco Intervertebral/terapia , Núcleo Pulposo/metabolismo , Dor nas Costas/terapia , Materiais Biocompatíveis/uso terapêutico , Humanos , Disco Intervertebral/fisiopatologia , Degeneração do Disco Intervertebral/genética , Núcleo Pulposo/patologia , Engenharia Tecidual
17.
Med Sci Monit ; 25: 4892-4900, 2019 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-31263091

RESUMO

BACKGROUND Intervertebral disc degeneration (IDD) is associated with low back and neck pain, but the mechanisms underlying its pathogenesis are unclear. In this study, we explored the function of microRNA-149 (miR-149) in inflammatory response mediated by lipopolysaccharide (LPS) in nucleus pulposus (NP) cells. MATERIAL AND METHODS Quantitative real-time PCR was used to detect miRNA and mRNA levels, while Western blotting was utilized to determine protein levels. ELISA was used to examine chemokine production. The correlation between miR-149 and MyD88 was assessed by reporter assay. Apoptosis was examined by flow cytometry. RESULTS miR-149 expression was significantly decreased after LPS exposure in NP cells. Overexpression of miR-149 reversed LPS-induced inhibition in aggrecan and collagen II expression and attenuated LPS-mediated promotion in the levels of MMP3, ADAMTS4, and inflammatory cytokines. Moreover, we found that miR-149 exerted its function by targeting MyD88 in NP cells. CONCLUSIONS miR-149 can inhibit the inflammatory response mediated by LPS in NP cells, and might be a potential target for the treatment of IDD.


Assuntos
MicroRNAs/genética , Fator 88 de Diferenciação Mieloide/metabolismo , Núcleo Pulposo/patologia , Animais , Apoptose/fisiologia , Citocinas/metabolismo , Matriz Extracelular/metabolismo , Disco Intervertebral/metabolismo , Disco Intervertebral/patologia , Degeneração do Disco Intervertebral/genética , Degeneração do Disco Intervertebral/metabolismo , Degeneração do Disco Intervertebral/patologia , Lipopolissacarídeos/metabolismo , MicroRNAs/biossíntese , Fator 88 de Diferenciação Mieloide/genética , Núcleo Pulposo/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley
18.
Gynecol Endocrinol ; 35(12): 1037-1039, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31274036

RESUMO

Perrault syndrome is a rare autosomal recessive disorder that affects both males and females. The syndrome causes deafness in males, however females display gonadal dysgenesis along with sensorineural hearing loss. Herein, we present a 27-year-old female patient who is deaf and mute along with primary amenorrhea. Hormonal assays revealed hypergonadotropic hypogonadism and the karyotype was 46 XX. Pelvic ultrasound described a hypoplastic uterus and streak ovaries. MRI of the spine showed degenerative discs and Tarlov cysts. Whole exome sequencing identified a LARS2 mutation and the patient was diagnosed with Perrault syndrome type four (PRLTS4).


Assuntos
Disgenesia Gonadal 46 XX/diagnóstico , Perda Auditiva Neurossensorial/diagnóstico , Adulto , Amenorreia/genética , Aminoacil-tRNA Sintetases/genética , Surdez/genética , Feminino , Disgenesia Gonadal 46 XX/genética , Disgenesia Gonadal 46 XX/fisiopatologia , Perda Auditiva Neurossensorial/genética , Perda Auditiva Neurossensorial/fisiopatologia , Humanos , Hipogonadismo/genética , Infertilidade Feminina/genética , Degeneração do Disco Intervertebral/diagnóstico por imagem , Degeneração do Disco Intervertebral/genética , Imagem por Ressonância Magnética , Cistos de Tarlov/diagnóstico por imagem , Cistos de Tarlov/genética , Ultrassonografia , Útero/diagnóstico por imagem
19.
Life Sci ; 232: 116565, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31251999

RESUMO

HEADINGS AIMS: The present study determined whether nucleus pulposus (NP) cells express hypoxia-inducible factor-2alpha (HIF-2α) and assessed its role in regulating the expression of catabolic factors during intervertebral disc degeneration. MATERIALS AND METHODS: Human degenerated NP tissues were acquired to examine the HIF-2α expression levels using immunohistochemistry, western blotting, and Real-time PCR. Human NP cells were cultivated under normoxic or hypoxic conditions, and the HIF-2α expression was determined. Then, human NP cells were treated with HIF-2α plasmids, HIF-2α siRNA, and tumor necrosis factor-alpha (TNF-α) to evaluate the role of HIF-2α in regulating matrix metalloproteinase (MMP) and aggrecanase expression. An in vivo rabbit disc degeneration model was established to demonstrate that HIF-2α plays a critical role in disc degeneration. KEY FINDINGS: We found that HIF-2α had a markedly elevated expression in human degenerated discs in the Grade III stage. HIF-2α protein and gene transcript levels in vitro were relatively higher under hypoxic conditions. The expression of MMP-13, ADAMTS-4 was decreased significantly in HIF-2α silencing condition, while the over-expression resulted in significantly increased levels of MMP-13 and ADAMTS-4. When cytokine TNF-α was added, HIF-2α was induced by nuclear factor-κB (NF-κB). The in vivo experiments showed that the HIF-2α controlled the catabolic factors MMP-13 and ADAMTS-4 that regulated the collagen II and aggrecan metabolism in disc degeneration. SIGNIFICANCE: HIF-2α is a catabolic regulator in disc degeneration and directly controls the catabolic genes. The suppression of HIF-2α expression leads to decelerates extracellular matrix degradation that might represent a therapeutic target for the degenerative disc.


Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/biossíntese , Degeneração do Disco Intervertebral/metabolismo , Adulto , Idoso , Agrecanas/metabolismo , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Citocinas/metabolismo , Endopeptidases/metabolismo , Feminino , Humanos , Hipóxia/metabolismo , Disco Intervertebral/metabolismo , Disco Intervertebral/patologia , Degeneração do Disco Intervertebral/genética , Degeneração do Disco Intervertebral/patologia , Masculino , Metaloproteinases da Matriz/metabolismo , Pessoa de Meia-Idade , Núcleo Pulposo/metabolismo , Núcleo Pulposo/patologia , Coelhos , Transdução de Sinais/fisiologia , Transcriptoma , Fator de Necrose Tumoral alfa/metabolismo
20.
Genes (Basel) ; 10(6)2019 06 07.
Artigo em Inglês | MEDLINE | ID: mdl-31181696

RESUMO

Two FGF4 retrogenes on chromosomes 12 (12-FGF4RG) and 18 (18-FGF4RG) contribute to short-limbed phenotypes in dogs. 12-FGF4RG has also been associated with intervertebral disc disease (IVDD). Both of these retrogenes were found to be widespread among dog breeds with allele frequencies ranging from 0.02 to 1; however, their additive contribution to disease is unknown. Surgical cases of IVDD (n = 569) were evaluated for age of onset, disc calcification, and genotypes for the FGF4 retrogenes. Multivariable linear regression analysis identified the presence of one or two copies of 12-FGF4RG associated with significantly younger age at first surgery in a dominant manner. 18-FGF4RG had only a minor effect in dogs with one copy. Multivariable logistic regression showed that 12-FGF4RG had an additive effect on radiographic disc calcification, while 18-FGF4RG had no effect. Multivariable logistic regression using mixed breed cases and controls identified only 12-FGF4RG as highly associated with disc herniation in a dominant manner (Odds Ratio, OR, 18.42, 95% Confidence Interval (CI) 7.44 to 50.26; P < 0.001). The relative risk for disc surgery associated with 12-FGF4RG varied from 5.5 to 15.1 within segregating breeds and mixed breeds. The FGF4 retrogene on CFA12 acts in a dominant manner to decrease the age of onset and increase the overall risk of disc disease in dogs. Other modifiers of risk may be present within certain breeds, including the FGF4 retrogene on CFA18.


Assuntos
Doenças do Cão/genética , Fator 4 de Crescimento de Fibroblastos/genética , Predisposição Genética para Doença , Degeneração do Disco Intervertebral/genética , Deslocamento do Disco Intervertebral/genética , Animais , Cruzamento , Doenças do Cão/fisiopatologia , Cães , Frequência do Gene , Genótipo , Disco Intervertebral/fisiopatologia , Degeneração do Disco Intervertebral/fisiopatologia , Deslocamento do Disco Intervertebral/fisiopatologia , Fenótipo
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