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1.
Zhonghua Er Ke Za Zhi ; 58(2): 135-139, 2020 Feb 02.
Artigo em Chinês | MEDLINE | ID: mdl-32102151

RESUMO

Objective: To explore the genotypes and phenotypes of osteogenesis imperfecta (OI) in Xinjiang Uygur children. Methods: The history of nine Uygur children with OI who were hospitalized in First Affiliated Hospital of Xinjiang Medical University from January 2013 to December 2017 were retrospectively reviewed. They were classified into 4 types according to the classical Sillence classification. The genes associated with OI were detected, and the pathogenic variation was assessed by InterVar and Alamut software according to the American College of Medical Genetics and Genomics (ACMG) recommendations. The phenotypes of children with different genotypes were further analyzed. Results: Nine cases aged 3 years and 6 monthes to 15 years were all clinically diagnosed as OI, the clinical manifes tations were repeated fractures, skeletal deformities,short stature, blue sclera, abnormol hearing, hypoplasia of dentin, and relaxation of Joint ligaments, among whom 6 was type Ⅲ OI, 3 were type Ⅳ OI. Nine mutations in 3 genes (COL1A1, COL1A2, and SERPINF1) were detected, and 5 of them were first reported and were all pathogenic variations. Conclusions: The cinical phenotypes of osteogenesis imperfecta in Xinjiang Uygur are complex and varied, but all of them have fractures and skeletal deformities. Genotype is different from that reported at China and abroad, and the SERPINF1 gene may have a higher incidence in Uyghur population. The genetic heterogeneity and unique gene variation pedigree of Uyghur osteogenesis imperfecta defects further provide a basis for the correlation between genotype and phenotype of osteogenesis defects.


Assuntos
Colágeno Tipo I/genética , Proteínas do Olho/genética , Fatores de Crescimento Neural/genética , Osteogênese Imperfeita/genética , Serpinas/genética , Adolescente , Densidade Óssea/genética , Criança , Pré-Escolar , China , Feminino , Predisposição Genética para Doença , Genótipo , Humanos , Lactente , Masculino , Mutação , Osteogênese Imperfeita/classificação , Osteogênese Imperfeita/diagnóstico , Osteogênese Imperfeita/patologia , Fenótipo , Estudos Retrospectivos
2.
J Stroke Cerebrovasc Dis ; 29(1): 104502, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31744764

RESUMO

BACKGROUND: Cerebral dopamine neurotrophic factor plays a critical role in repairing and maintaining healthy neurons in pathological conditions such as stroke. However, the association between cerebral dopamine neurotrophic factor expression and stroke has only recently been investigated in preclinical models and is rarely described in human studies. OBJECTIVES: The aims of this were to examine neurological alterations mirrored in human blood platelet cerebral dopamine neurotrophic factor gene expression. Cerebral dopamine neurotrophic factor is expressed in both the central nervous system and peripheral blood. Blood platelets are often used to model neuronal behavior because they exhibit biochemical impairments similar to brain tissues of patients with neurological disorders. METHODS: RNA was isolated from platelets and cDNA was synthesized to quantify cerebral dopamine neurotrophic factor gene expression of 36 stroke patients compared to 72 healthy aged-matched controls through real-time PCR. Further grouping analyses of data with regard to age, sex, and medication history were performed. RESULTS: Cerebral dopamine neurotrophic factor gene expression was significantly reduced in stroke patients relative to control subjects (P = .013). Subsequent analysis revealed a significant difference in expression between males and females within the control group (P = .026). Decreased cerebral dopamine neurotrophic factor expression was only observed in male stroke patients compared to their sex-matched controls (P = .008). Grouping stroke patients based on their medication history did not significantly alter cerebral dopamine neurotrophic factor gene expression. CONCLUSIONS: Further studies investigating cerebral dopamine neurotrophic factor expression could be directed towards the interplay of the central nervous system, hematopoietic derivatives, and utilizing cerebral dopamine neurotrophic factor as a therapeutic tool.


Assuntos
Plaquetas/metabolismo , Fatores de Crescimento Neural/sangue , Acidente Vascular Cerebral/sangue , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores/sangue , Estudos de Casos e Controles , Regulação para Baixo , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fatores de Crescimento Neural/genética , RNA Mensageiro/sangue , Fatores Sexuais , Acidente Vascular Cerebral/diagnóstico , Acidente Vascular Cerebral/genética , Adulto Jovem
3.
PLoS Comput Biol ; 15(11): e1007460, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31682594

RESUMO

Radiation therapy is an important and effective treatment option for prostate cancer, but high-risk patients are prone to relapse due to radioresistance of cancer cells. Molecular mechanisms that contribute to radioresistance are not fully understood. Novel computational strategies are needed to identify radioresistance driver genes from hundreds of gene copy number alterations. We developed a network-based approach based on lasso regression in combination with network propagation for the analysis of prostate cancer cell lines with acquired radioresistance to identify clinically relevant marker genes associated with radioresistance in prostate cancer patients. We analyzed established radioresistant cell lines of the prostate cancer cell lines DU145 and LNCaP and compared their gene copy number and expression profiles to their radiosensitive parental cells. We found that radioresistant DU145 showed much more gene copy number alterations than LNCaP and their gene expression profiles were highly cell line specific. We learned a genome-wide prostate cancer-specific gene regulatory network and quantified impacts of differentially expressed genes with directly underlying copy number alterations on known radioresistance marker genes. This revealed several potential driver candidates involved in the regulation of cancer-relevant processes. Importantly, we found that ten driver candidates from DU145 (ADAMTS9, AKR1B10, CXXC5, FST, FOXL1, GRPR, ITGA2, SOX17, STARD4, VGF) and four from LNCaP (FHL5, LYPLAL1, PAK7, TDRD6) were able to distinguish irradiated prostate cancer patients into early and late relapse groups. Moreover, in-depth in vitro validations for VGF (Neurosecretory protein VGF) showed that siRNA-mediated gene silencing increased the radiosensitivity of DU145 and LNCaP cells. Our computational approach enabled to predict novel radioresistance driver gene candidates. Additional preclinical and clinical studies are required to further validate the role of VGF and other candidate genes as potential biomarkers for the prediction of radiotherapy responses and as potential targets for radiosensitization of prostate cancer.


Assuntos
Perfilação da Expressão Gênica/métodos , Recidiva Local de Neoplasia/genética , Tolerância a Radiação/genética , Apoptose , Biomarcadores Tumorais/genética , Linhagem Celular Tumoral , Biologia Computacional/métodos , Dosagem de Genes/genética , Regulação Neoplásica da Expressão Gênica/genética , Redes Reguladoras de Genes/genética , Inativação Gênica , Humanos , Masculino , Fatores de Crescimento Neural/genética , Neoplasias da Próstata/genética , RNA Mensageiro/genética , RNA Interferente Pequeno , Recidiva , Transdução de Sinais/genética , Transcriptoma/genética
4.
Biosci Biotechnol Biochem ; 83(12): 2345-2354, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31524073

RESUMO

The effect of Lactobacillus plantarum SNK12 (CPLP) supplementation on mRNA levels of hippocampal neurotrophic factors and gamma aminobutyric acid receptors (GABAR) was tested. In Experiment 1, stress-free, unsupplemented and CPLP (4 × 108 cells/head)-supplemented male C57BL/6J (B6) mice were the experimental animals. In Experiment 2, intruder (male, B6) mice [negative control; unsupplemented, sub-chronic mild social defeat stress (sCSDS)-induced; and CPLP-supplemented, sCSDS-induced] were exposed to aggressor mice (adult male Slc:ICR). mRNA levels of neurotrophic factors and GABAR in hippocampal samples of these mice were analyzed. In CPLP-supplemented mice of both experiments, mRNA levels of bdnf, nt-3, and GABAR were upregulated. Moreover, a tendency toward the improvement of habituation ability (Experiment 1) and behavior (Experiment 2) was observed in mice, which may be associated with upregulated neurotrophic factors and GABAR. We demonstrated that oral supplementation of CPLP to stress-free and stress-induced mice upregulated mRNA levels of hippocampal neurotrophic factors and GABAR.


Assuntos
Fator Neurotrófico Derivado do Encéfalo/genética , Hipocampo/metabolismo , Lactobacillus plantarum , Fatores de Crescimento Neural/genética , Probióticos , RNA Mensageiro/genética , Receptores de GABA/genética , Estresse Psicológico , Animais , Comportamento Animal , Peso Corporal , Comportamento de Ingestão de Líquido , Comportamento Alimentar , Masculino , Camundongos , Camundongos Endogâmicos C57BL
5.
Int J Mol Sci ; 20(17)2019 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-31484425

RESUMO

Glaucoma and other optic neuropathies are characterized by axonal transport deficits. Axonal cargo travels back and forth between the soma and the axon terminus, a mechanism ensuring homeostasis and the viability of a neuron. An example of vital molecules in the axonal cargo are neurotrophic factors (NTFs). Hindered retrograde transport can cause a scarcity of those factors in the retina, which in turn can tilt the fate of retinal ganglion cells (RGCs) towards apoptosis. This postulation is one of the most widely recognized theories to explain RGC death in the disease progression of glaucoma and is known as the NTF deprivation theory. For several decades, research has been focused on the use of NTFs as a novel neuroprotective glaucoma treatment. Until now, results in animal models have been promising, but translation to the clinic has been highly disappointing. Are we lacking important knowledge to lever NTF therapies towards the therapeutic armamentarium? Or did we get the wrong end of the stick regarding the NTF deprivation theory? In this review, we will tackle the existing evidence and caveats advocating for and against the target-derived NTF deprivation theory in glaucoma, whilst digging into associated therapy efforts.


Assuntos
Fatores de Crescimento Neural/metabolismo , Células Ganglionares da Retina/metabolismo , Animais , Glaucoma/metabolismo , Glaucoma/patologia , Humanos , Fatores de Crescimento Neural/genética , Nervo Óptico/metabolismo , Doenças do Nervo Óptico/metabolismo , Retina/metabolismo , Células Ganglionares da Retina/citologia
6.
Biomed Pharmacother ; 118: 109263, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31369988

RESUMO

It is well known that chamomile is one of the oldest known medicinal herbs and has been used to treat various disorders, but it is mainly German chamomile. The effects of Roman chamomile on depression still unclear. In this study, we used chronically stressed mice to investigate whether inhalation of Roman chamomile essential oil affects depression-like behavior. We previously reported that restraint and water immersion stress produce depression-like behavior and a blunted response to the tricyclic antidepressant clomipramine. Each mouse was exposed to restraint and water immersion stress for 15 days, and resistance to the effect of clomipramine was induced in a behavioral despair paradigm. In the present study, we found that cotreatment with clomipramine and inhalation of Roman chamomile attenuated depression-like behavior in a forced swim test. Next, we examined the hippocampal mRNA levels of two cytokines, tumor necrosis factor (TNF) alpha and interleukin-6 (IL-6); a neurotrophic factor, brain derived-neurotrophic factor (BDNF); and nerve growth factor (NGF). TNF alpha, IL-6 and BDNF mRNA levels did not change in the hippocampus of stressed mice. However, the NGF mRNA level was significantly decreased, and this decrease was not attenuated by treatment with clomipramine or inhalation of Roman chamomile alone. We also examined whether Roman chamomile combined with clomipramine treatment affects hippocampal neurogenesis and serum corticosterone levels. Stressed mice had fewer doublecortin (DCX)-positive cells in the subgranular zone of the dentate gyrus, but this was significantly attenuated by Roman chamomile and clomipramine treatment. In addition, the serum corticosterone level was also significantly decreased by treatment with Roman chamomile and clomipramine. These results suggest that Roman chamomile inhalation may enhance the antidepressant effect of clomipramine by increasing hippocampal neurogenesis and modulating corticosterone levels in patients with treatment-resistant depression.


Assuntos
Comportamento Animal , Chamaemelum/química , Clomipramina/uso terapêutico , Depressão/tratamento farmacológico , Exposição por Inalação , Extratos Vegetais/uso terapêutico , Animais , Proliferação de Células/efeitos dos fármacos , Clomipramina/farmacologia , Corticosterona/sangue , Citocinas/genética , Citocinas/metabolismo , Depressão/sangue , Quimioterapia Combinada , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Fatores de Crescimento Neural/genética , Fatores de Crescimento Neural/metabolismo , Neurogênese/efeitos dos fármacos , Extratos Vegetais/farmacologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Estresse Psicológico/sangue , Estresse Psicológico/tratamento farmacológico
7.
Invest Ophthalmol Vis Sci ; 60(8): 2888-2894, 2019 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-31266060

RESUMO

Purpose: Extremely preterm infants are at increased risk for retinopathy of prematurity (ROP). We previously identified several inflammatory proteins that were expressed early in life and are associated with an increased risk of ROP and several angiogenic and neurotrophic growth factors in the neonatal systemic circulation that are associated with a lower risk of ROP. In this paper, we report the results of a set of analyses designed to test the hypothesis that placental CpG methylation levels of 12 inflammation-, angiogenic-, and neurotrophic-associated genes predict the occurrence of prethreshold ROP in extremely preterm newborns. Methods: We used placental CpG methylation data from 395 newborns from the Extremely Low Gestational Age Newborns study. Results: Multivariable regression models revealed that placental DNA methylation of 16 CpG sites representing 8 genes were associated with prethreshold ROP. Specifically, CpG methylation in the serum amyloid A SAA1 and SAA2, brain-derived neurotrophic factor (BDNF), myeloperoxidase (MPO), C-reactive protein (CRP), angiopoietin 1 (ANGPT1), and tumor necrosis factor receptor superfamily member 1B (TNFRSF1B) genes was associated with a lower risk of prethreshold ROP. Conversely, CpG methylation at three probes within tumor necrosis factor receptor superfamily member 1A (TNFRSF1A) and in two alternative probes within the BDNF and ANGPT1 genes was associated with an increased risk of ROP. Conclusions: CpG methylation may be a useful marker for improving ROP prediction, opening the opportunity for early intervention to lessen disease severity.


Assuntos
Proteínas Angiogênicas/genética , Ilhas de CpG/fisiologia , Metilação de DNA , Inflamação/genética , Fatores de Crescimento Neural/genética , Placenta/metabolismo , Retinopatia da Prematuridade/genética , Adulto , Feminino , Idade Gestacional , Humanos , Recém-Nascido de Peso Extremamente Baixo ao Nascer , Lactente Extremamente Prematuro , Recém-Nascido , Peptídeos e Proteínas de Sinalização Intercelular/genética , Masculino , Razão de Chances , Gravidez , Fatores de Risco
8.
J Neuroinflammation ; 16(1): 133, 2019 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-31266507

RESUMO

BACKGROUND: Treatments for autoimmune diseases aim to dampen autoreactivity while preserving normal immune function. In CD4+ T cells, the transcription factor Oct1/Pou2f1 is a dispensable transcription factor for T cell development and response to primary infection, but promotes expression of target genes, including Il2 and Ifng, under conditions of antigen reencounter. As a result, they are more strongly expressed upon secondary stimulation. Such repeated antigen encounters occur in memory recall responses, in autoimmunity where self-antigen can be recognized multiple times, and in chronic infection where foreign antigen is persistent. Based on these previous findings, we hypothesized that Oct1 loss would protect animals from autoimmunity but maintain normal responses to pathogens in the CNS. OBJECTIVE: We used a conditional mouse Oct1 (Pou2f1) allele and a CD4-Cre driver to determine the effect of T cell-specific Oct1 loss on autoimmune- and viral-induced neuroinflammation using an autoantigen-driven EAE model of autoimmunity and a JHMV model of viral infection. RESULTS: Oct1 conditional deletion mitigated clinical scores and reduced infiltrating T cells and cytokine production in the EAE model. Consistently, Oct1-deficient CD4+ T cells stimulated in vitro showed increased expression of markers associated with T cell anergy, particularly in the absence of co-stimulatory signals. In contrast, anti-viral T cell effector functions are intact in the absence of Oct1, with no changes in neuroinflammation, infiltrating T cells or cytokine production. CONCLUSION: Our findings uncover a significant difference between the effect of Oct1 loss on autoimmune and anti-pathogen responses, which potentially could be exploited for therapeutic benefit.


Assuntos
Autoimunidade/fisiologia , Linfócitos T CD4-Positivos/metabolismo , Encefalomielite Autoimune Experimental/metabolismo , Mediadores da Inflamação/metabolismo , Fatores de Crescimento Neural/metabolismo , Fator 1 de Transcrição de Octâmero/deficiência , Sequência de Aminoácidos , Animais , Linfócitos T CD4-Positivos/imunologia , Encefalomielite Autoimune Experimental/genética , Encefalomielite Autoimune Experimental/imunologia , Inflamação/genética , Inflamação/imunologia , Inflamação/metabolismo , Mediadores da Inflamação/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Fatores de Crescimento Neural/genética , Fatores de Crescimento Neural/imunologia , Fator 1 de Transcrição de Octâmero/genética , Fator 1 de Transcrição de Octâmero/imunologia
9.
PLoS Genet ; 15(6): e1007960, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31233487

RESUMO

UNC-6/Netrin is a conserved axon guidance cue that directs growth cone migrations in the dorsal-ventral axis of C. elegans and in the vertebrate spinal cord. UNC-6/Netrin is expressed in ventral cells, and growth cones migrate ventrally toward or dorsally away from UNC-6/Netrin. Recent studies of growth cone behavior during outgrowth in vivo in C. elegans have led to a polarity/protrusion model in directed growth cone migration away from UNC-6/Netrin. In this model, UNC-6/Netrin first polarizes the growth cone via the UNC-5 receptor, leading to dorsally biased protrusion and F-actin accumulation. UNC-6/Netrin then regulates protrusion based on this polarity. The receptor UNC-40/DCC drives protrusion dorsally, away from the UNC-6/Netrin source, and the UNC-5 receptor inhibits protrusion ventrally, near the UNC-6/Netrin source, resulting in dorsal migration. UNC-5 inhibits protrusion in part by excluding microtubules from the growth cone, which are pro-protrusive. Here we report that the RHO-1/RhoA GTPase and its activator GEF RHGF-1 inhibit growth cone protrusion and MT accumulation in growth cones, similar to UNC-5. However, growth cone polarity of protrusion and F-actin were unaffected by RHO-1 and RHGF-1. Thus, RHO-1 signaling acts specifically as a negative regulator of protrusion and MT accumulation, and not polarity. Genetic interactions are consistent with RHO-1 and RHGF-1 acting with UNC-5, as well as with a parallel pathway, to regulate protrusion. The cytoskeletal interacting molecule UNC-33/CRMP was required for RHO-1 activity to inhibit MT accumulation, suggesting that UNC-33/CRMP might act downstream of RHO-1. In sum, these studies describe a new role of RHO-1 and RHGF-1 in regulation of growth cone protrusion by UNC-6/Netrin.


Assuntos
Proteínas de Caenorhabditis elegans/genética , Fatores de Troca do Nucleotídeo Guanina/genética , Netrinas/genética , Neurônios/metabolismo , Proteínas rho de Ligação ao GTP/genética , Animais , Orientação de Axônios/genética , Axônios/metabolismo , Caenorhabditis elegans/genética , Caenorhabditis elegans/crescimento & desenvolvimento , Movimento Celular/genética , Polaridade Celular/genética , Regulação da Expressão Gênica no Desenvolvimento/genética , Cones de Crescimento/metabolismo , Microtúbulos/genética , Fatores de Crescimento Neural/genética , Fenótipo , Pseudópodes/genética , Receptores de Superfície Celular/genética , Transdução de Sinais/genética
10.
J Genet ; 98(2)2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31204718

RESUMO

Mutations in several genes, including SERPRINF1 and COL1A1, have been associated with the development of osteogenesis imperfecta (OI). Here, we reported the co-occurrence of a rare heterozygous variant (c.167C>G p.Ala56Gly) in SERPRINF1 and a novel heterozygous mutation (c.1634G>A p.Gly545Asp) in COL1A1 in a foetus with a severe form of OI. Bioinformatics modelling revealed that the effect of the mutation on SPERINF1 is neutral. In contrast, the mutation in COL1A1 is deleterious. It is predicted to cause distortion of the α (1) chain of the type I collagen and results in structural instability of the protein. Therefore, a novel dominant variant of COL1A1 likely underlies the severe foetal pathology observed, although we do not exclude the possibility that the heterozygous mutations in SERPINF and COL1A1 may interact and co-ordinately cause pathogenesis. This novel COL1A1 mutation is recommended to be included in the diagnostic panels for OI.


Assuntos
Colágeno Tipo I/genética , Proteínas do Olho/genética , Heterozigoto , Mutação , Fatores de Crescimento Neural/genética , Osteogênese Imperfeita/genética , Serpinas/genética , Alelos , Sequência de Aminoácidos , Instabilidade Cromossômica , Colágeno Tipo I/química , Biologia Computacional/métodos , Análise Mutacional de DNA , Evolução Molecular , Proteínas do Olho/química , Feminino , Instabilidade Genômica , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Modelos Moleculares , Fatores de Crescimento Neural/química , Osteogênese Imperfeita/diagnóstico , Fenótipo , Gravidez , Conformação Proteica , Serpinas/química , Relação Estrutura-Atividade , Ultrassonografia
11.
Mol Med Rep ; 20(2): 1443-1450, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31173218

RESUMO

Autophagy is a conserved catabolic process by which cytoplasmic components are delivered into lysosomes for degradation. Pigment epithelium­derived factor (PEDF) has been reported to be associated with autophagy and can induce p53 expression; however, the mechanism relating PEDF with autophagy in endothelial cells remains poorly understood. The present study aimed to investigate the association between the PEDF­p53­sestrin pathway and autophagy in human umbilical vein endothelial cells (HUVECs). PEDF­induced autophagy was examined by fluorescence microscopy and western blot analysis. p53 small interfering (si)RNA and sestrin2 siRNA were constructed and transfected into HUVECs prior to PEDF treatment. The protein expression levels of microtubule­associated protein light chain 3 (LC3) I, LC3 II and p62 were evaluated by western blot analysis, and the mRNA expression levels of p53 and sestrin2 were determined using reverse transcription­quantitative polymerase chain reaction analysis. The regulation of mechanistic target of rapamycin (mTOR) was reflected by p70S6 kinase (p70S6K) and eukaryotic translation initiation factor 4E­binding protein 1 (4E­BP1) protein expression levels, as determined by western blot analysis. PEDF could induce HUVEC autophagy by sequentially inducing p53 and sestrin2 expression, as observed by fluorescence microscopy and western blot analysis. Conversely, the induction of sestrin2 by PEDF was eliminated by p53 siRNA. In addition, p53 siRNA and sestrin2 siRNA could attenuate PEDF­induced HUVEC autophagy. Inhibition of mTOR may be the mechanism responsible for PEDF­induced autophagy; as p70S6K and 4E­BP1 phosphorylation levels were significantly upregulated in p53 siRNA­treated and sestrin2 siRNA­treated groups. The findings of the present study indicated that PEDF may trigger autophagy in HUVECs by inducing p53 and sestrin2 expression, and inhibiting mTOR expression; these findings may contribute to the improved understanding of diseases, including cancer and atherosclerosis.


Assuntos
Autofagia/genética , Proteínas do Olho/genética , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Fatores de Crescimento Neural/genética , Proteínas Nucleares/genética , Serpinas/genética , Transdução de Sinais/genética , Proteína Supressora de Tumor p53/genética , Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Autofagia/efeitos dos fármacos , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Proteínas do Olho/metabolismo , Proteínas do Olho/farmacologia , Regulação da Expressão Gênica , Células Endoteliais da Veia Umbilical Humana/citologia , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Proteínas Associadas aos Microtúbulos/genética , Proteínas Associadas aos Microtúbulos/metabolismo , Fatores de Crescimento Neural/metabolismo , Fatores de Crescimento Neural/farmacologia , Proteínas Nucleares/antagonistas & inibidores , Proteínas Nucleares/metabolismo , Fosforilação/efeitos dos fármacos , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Proteínas Quinases S6 Ribossômicas 70-kDa/genética , Proteínas Quinases S6 Ribossômicas 70-kDa/metabolismo , Proteína Sequestossoma-1/genética , Proteína Sequestossoma-1/metabolismo , Serpinas/metabolismo , Serpinas/farmacologia , Serina-Treonina Quinases TOR/genética , Serina-Treonina Quinases TOR/metabolismo , Proteína Supressora de Tumor p53/antagonistas & inibidores , Proteína Supressora de Tumor p53/metabolismo
12.
Int J Mol Sci ; 20(11)2019 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-31159418

RESUMO

Human tonsil-derived mesenchymal stem cells (T-MSCs) are newly identified MSCs and present typical features of MSCs, including having the differentiation capacity into the three germ layers and excellent proliferation capacity. They are easily sourced and are useful for stem cell therapy in various disease states. We previously reported that T-MSCs could be differentiated into skeletal myocytes and Schwann-like cells; therefore, they are a promising candidate for cell therapies for neuromuscular disease. Motor neurons (MNs), which regulate spontaneous behavior, are affected by a wide range of MN diseases (MNDs) for which there are no effective remedies. We investigated the differentiation potential of MN-like cells derived from T-MSCs (T-MSC-MNCs) for application to therapy of MNDs. After the process of MN differentiation, the expression of MN-related markers, including Islet 1, HB9/HLXB9 (HB9), and choline acetyltransferase (ChAT), was increased when compared with undifferentiated T-MSCs. The secretion of acetylcholine to the conditioned medium was significantly increased after MN differentiation. We cocultured T-MSC-MNCs and human skeletal muscle cells, and confirmed the presence of the acetylcholine receptor clusters, which demonstrated the formation of neuromuscular junctions. The potential functional improvements afforded by these T-MSC-MNCs could be useful in the treatment of MNDs caused by genetic mutation, viral infection, or environmental problems.


Assuntos
Diferenciação Celular , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Neurônios Motores/citologia , Neurônios Motores/fisiologia , Junção Neuromuscular/fisiologia , Tonsila Palatina/citologia , Acetilcolina/metabolismo , Biomarcadores , Células Cultivadas , Expressão Gênica , Humanos , Imuno-Histoquímica , Fibras Musculares Esqueléticas/metabolismo , Fatores de Crescimento Neural/genética , Fatores de Crescimento Neural/metabolismo , Células-Tronco Neurais/citologia , Células-Tronco Neurais/metabolismo
13.
Leg Med (Tokyo) ; 38: 83-91, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31108272

RESUMO

Genes differentially expressed after death were selected to construct a mathematical model for early postmortem interval estimation. Sprague Dawley rats were sacrificed and placed at temperatures of 4 °C, 15 °C, 25 °C, and 35 °C. Brain tissues were collected at 0, 6, 12, 18, and 24 h after death and total RNA was extracted. Changes in gene transcript levels after death were detected using microarray expression profiling and differentially expressed genes was screened. Expanded experiments were performed to validate gene transcript levels at different temperatures using the reverse transcription real-time quantitative polymerase chain reaction. Six genes with high coefficients of determination were chosen for construction of mathematical models. Optimal ternary cubic equations were built using R software with temperature, postmortem interval and ΔCq defined as the independent variable x, y and z, respectively. Equations were converted into a three-dimensional visual statistical model using MATLAB. Animal samples were used to validate the mathematical models. Results showed that the 5srRNA showed best stability at four temperatures. The genes Ninj2, Grifin, Arpp19, and Hopx showed high coefficients of determination (>80%) and low error (<3h) in verification experiments which indicate that they are potential markers for early postmortem interval estimation.


Assuntos
Medicina Legal/métodos , Expressão Gênica , Mudanças Depois da Morte , RNA/genética , RNA/isolamento & purificação , Transcrição Genética , Animais , Biomarcadores , Moléculas de Adesão Celular Neuronais/genética , Moléculas de Adesão Celular Neuronais/metabolismo , Proteínas do Olho/genética , Proteínas do Olho/metabolismo , Galectinas/genética , Galectinas/metabolismo , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Análise em Microsséries , Modelos Teóricos , Fatores de Crescimento Neural/genética , Fatores de Crescimento Neural/metabolismo , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Temperatura Ambiente , Fatores de Tempo , Proteínas Supressoras de Tumor/genética , Proteínas Supressoras de Tumor/metabolismo
14.
BMC Cancer ; 19(1): 484, 2019 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-31117965

RESUMO

BACKGROUND: Metastases account for 90% of all cancer-related deaths, becoming a therapeutic problem. Approximately 50% of all uveal melanoma (UM) patients will develop metastases, mainly in the liver. Post-mortem analyses of livers from metastatic UM patients showed two different metastatic growth patterns: infiltrative and nodular. The infiltrative pattern exhibits tumor infiltration directly to the hepatic lobule and minimal angiogenesis. The nodular pattern shows clusters of tumor cells around the portal venules that efface the liver parenchyma. We recently demonstrated Natural Killer (NK) cells play a pivotal role in the control of hepatic metastases and the pigment epithelial-derived factor (PEDF) controls angiogenesis in the liver using our established ocular melanoma animal model. In this study we investigated the role of NK cells and PEDF in the development of metastatic growth patterns, as this can contribute to the development of novel therapeutics specific towards each growth pattern. METHODS: We utilize our established ocular melanoma animal model by inoculation of B16-LS9 melanoma cells into C57BL/6 J mice (WT), anti-asialo GM1-treated C57BL/6 J mice (NK-depleted), and PEDF-/- C57BL/6 J mice. Three weeks after inoculation we evaluated the metastatic growth patterns and stratified them based of the numbers of tumor cells. To evaluate angiogenesis the mean vascular density (MVD) was calculated. The immune compartment of the liver was analyzed by flow cytometry. RESULTS: Our in vivo work showed two distinct metastatic growth patterns, the infiltrative and nodular, recapitulating the post-mortem analyses on human liver tissue. We discovered NK cells control the infiltrative growth. In contrast, PEDF controlled anti-angiogenic responses, showing higher MVD values compared to NK-depleted and WT animals. The myeloid lineage, comprised of monocytes, macrophages, and myeloid-derived suppressor cells, was reduced in the absence of NK cells or PEDF. CONCLUSIONS: Our animal model recapitulates the metastatic growth patterns observed in the human disease. We demonstrated a role for NK cells in the development of the infiltrative growth pattern, and a role for PEDF in the development of the nodular pattern. The understanding of the complexity associated with the metastatic progression has profound clinical implications in the diagnostic and disease-management as we can develop and direct more effective therapies.


Assuntos
Anticorpos/farmacologia , Proteínas do Olho/genética , Células Matadoras Naturais/efeitos dos fármacos , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/secundário , Melanoma/imunologia , Fatores de Crescimento Neural/genética , Serpinas/genética , Neoplasias Uveais/imunologia , Animais , Linhagem Celular Tumoral , Proteínas do Olho/metabolismo , Feminino , Gangliosídeo G(M1)/antagonistas & inibidores , Técnicas de Inativação de Genes , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/imunologia , Macrófagos/metabolismo , Melanoma/genética , Melanoma/patologia , Camundongos , Camundongos Endogâmicos C57BL , Monócitos/metabolismo , Células Supressoras Mieloides/metabolismo , Transplante de Neoplasias , Fatores de Crescimento Neural/metabolismo , Serpinas/metabolismo , Neoplasias Uveais/genética , Neoplasias Uveais/patologia
15.
J Clin Pathol ; 72(7): 460-467, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31072837

RESUMO

The neurotrophic tyrosine receptor kinase (NTRK) gene family encodes three tropomyosin receptor kinases (TRKA, TRKB, TRKC) that contribute to central and peripheral nervous system development and function. NTRK gene fusions are oncogenic drivers of various adult and paediatric tumours. Several methods have been used to detect NTRK gene fusions including immunohistochemistry, fluorescence in situ hybridisation, reverse transcriptase polymerase chain reaction, and DNA- or RNA-based next-generation sequencing. For patients with TRK fusion cancer, TRK inhibition is an important therapeutic target. Following the FDA approval of the selective TRK inhibitor, larotrectinib, as well as the ongoing development of multi-kinase inhibitors with activity in TRK fusion cancer, testing for NTRK gene fusions should become part of the standard diagnostic process. In this review we discuss the biology of NTRK gene fusions, and we present a testing algorithm to aid detection of these gene fusions in clinical practice and guide treatment decisions.


Assuntos
Neoplasias/genética , Fatores de Crescimento Neural/genética , Proteínas de Fusão Oncogênica/genética , Inibidores de Proteínas Quinases/uso terapêutico , Pirazóis/uso terapêutico , Pirimidinas/uso terapêutico , Receptores Proteína Tirosina Quinases/genética , Algoritmos , Humanos , Glicoproteínas de Membrana/antagonistas & inibidores , Glicoproteínas de Membrana/genética , Neoplasias/tratamento farmacológico , Neoplasias/enzimologia , Fatores de Crescimento Neural/antagonistas & inibidores , Receptores Proteína Tirosina Quinases/antagonistas & inibidores , Receptor trkA/antagonistas & inibidores , Receptor trkA/genética , Receptor trkB/antagonistas & inibidores , Receptor trkB/genética , Receptor trkC/antagonistas & inibidores , Receptor trkC/genética
16.
Int J Oncol ; 55(1): 116-130, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31059004

RESUMO

Triple­negative breast cancer (TNBC) cells form angiogenesis­independent vessel­like structures to survive, known as vasculogenic mimicry (VM), contributing to a poor prognosis for cancer patients. Nuclear localized class I histone deacetylases (HDACs) enzymes, particularly HDACs 1, 2, 3 deacetylate chromatin histones, are overexpressed in cancers and epigenetically regulate the expression of genes involved in cancer initiation and progression. The specific HDAC inhibitor, entinostat, has been shown to attenuate tumor progression and metastasis in TNBC. In this study, we hypothesized that entinostat would enhance the expression of anti­angiogenic and tumor suppressor genes and would thus suppress VM structures in TNBC cells in a 3D Matrigel cell culture preclinical model. Our data indicated that invasive triple­negative MDA­MB­231, LM2­4 and BT­549 breast cancer cells, but not poorly invasive luminal MCF­7 cells, efficiently underwent matrix­associated VM formation. Approximately 80% of TNBC cells with the stem cell phenotype potential formed vessel­like structures when mixed with Matrigel and cultured in the low attachment tissue culture plate. The molecular mechanisms of VM formation are rather complex, while angiogenesis inhibitor genes are downregulated and pro­angiogenesis genes are upregulated in VM­forming cells. Our data revealed that treatment of the TNBC VM phenotype cells with entinostat epigenetically led to the re­expression of the anti­angiogenic genes, serpin family F member 1 (SERPINF1) and thrombospondin 2 (THBS2), and to that of the tumor suppressor genes, phosphatase and tensin homolog (PTEN) and p21, and reduced VM structures. We also found that treatment of the TNBC VM phenotype cells with entinostat downregulated the expression of vascular endothelial growth factor A (VEGF­A), and that of the epithelial­mesenchymal transition (EMT)­related genes, Vimentin and ß­catenin. METABIRC and TCGA breast cancer cohort mRNA expression data analysis revealed that a high expression of the anti­angiogenesis­associated genes, THBS2, SERPINF1 and serpin family B member 5 (SERPINB5), and of the tumor suppressor gene, PTEN, was associated with a better overall survival (OS) of breast cancer patients. Taken together, the findings of this study demonstrate that HDACs 1, 2, 3 partly contribute to VM formation in TNBC cells; thus, HDACs may be an important therapeutic target for TNBC.


Assuntos
Inibidores da Angiogênese/genética , Benzamidas/farmacologia , Genes Supressores de Tumor/efeitos dos fármacos , Histona Desacetilase 1/antagonistas & inibidores , Inibidores de Histona Desacetilases/farmacologia , Piridinas/farmacologia , Neoplasias de Mama Triplo Negativas/genética , Linhagem Celular Tumoral , Epigênese Genética , Proteínas do Olho/genética , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Células MCF-7 , Fatores de Crescimento Neural/genética , Serpinas/genética , Análise de Sobrevida , Trombospondinas/genética , Neoplasias de Mama Triplo Negativas/tratamento farmacológico
17.
Artigo em Inglês | MEDLINE | ID: mdl-31128281

RESUMO

Reserpine is widely used for treatment of hypertension and schizophrenia. As a specific inhibitor of monoamine transporters, reserpine is known to deplete monoamine neurotransmitters and cause decreased movement symptoms. However, how zebrafish larvae respond to reserpine treatment is not well studied. Here we show that swimming distance and average velocity are significantly reduced after reserpine exposure under various stimulatory conditions. Using liquid chromatograph-mass spectrometer analysis, decreased levels of monoamines (e.g. dopamine, noradrenaline, and serotonin) were detected in reserpine-treated larvae. Moreover, reserpine treatment significantly reduced the number of dopaminergic neurons, which was identified with th (Tyrosine Hydroxylase) in situ hybridization in the preoptic area. Interestingly, dopaminergic neuron development-associated genes, such as otpa, otpb, wnt1, wnt3, wnt5 and manf, were downregulated in reserpine treated larvae. Our data indicates that 2 mg/L reserpine exposure induces dopaminergic neuron damage in the brain, demonstrating a chemical induced depression-like model in zebrafish larvae for future drug development.


Assuntos
Neurônios Dopaminérgicos/efeitos dos fármacos , Larva/efeitos dos fármacos , Síndromes Neurotóxicas/etiologia , Reserpina/toxicidade , Peixe-Zebra , Animais , Monoaminas Biogênicas/metabolismo , Neurônios Dopaminérgicos/patologia , Embrião não Mamífero/efeitos dos fármacos , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Larva/metabolismo , Luz , Locomoção/efeitos dos fármacos , Fatores de Crescimento Neural/genética , Som , Proteína Wnt-5a/genética , Proteína Wnt1/genética , Proteína Wnt3A/genética , Peixe-Zebra/embriologia , Peixe-Zebra/genética , Proteínas de Peixe-Zebra/genética
18.
PLoS One ; 14(5): e0216987, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31091274

RESUMO

Chronic pressure overload due to aortic valve stenosis leads to pathological cardiac hypertrophy and heart failure. Hypertrophy is accompanied by an increase in myocyte surface area, which requires a proportional increase in the number of cell-cell and cell-matrix contacts to withstand enhanced workload. In a proteomic analysis we identified nerve injury-induced protein 1 (Ninjurin1), a 16kDa transmembrane cell-surface protein involved in cell adhesion and nerve repair, to be increased in hypertrophic hearts from patients with aortic stenosis. We hypothesised that Ninjurin1 is involved in myocyte hypertrophy. We analyzed cardiac biopsies from aortic-stenosis patients and control patients undergoing elective heart surgery. We studied cardiac hypertrophy in mice after transverse aortic constriction and angiotensin II infusions, and performed mechanistic analyses in cultured myocytes. We assessed the physiological role of ninjurin1 in zebrafish during heart and skeletal muscle development. Ninjurin1 was increased in hearts of aortic stenosis patients, compared to controls, as well as in hearts from mice with cardiac hypertrophy. Besides the 16kDa Ninjurin1 (Ninjurin1-16) we detected a 24kDa variant of Ninjurin1 (Ninjurin1-24), which was predominantly expressed during myocyte hypertrophy. We disclosed that the higher molecular weight of Ninjurin1-24 was caused by N-glycosylation. Ninjurin1-16 was contained in the cytoplasm of myocytes where it colocalized with stress-fibers. In contrast, Ninjurin1-24 was localized at myocyte membranes. Gain and loss-of-function experiments showed that Ninjurin1-24 plays a role in myocyte hypertrophy and myogenic differentiation in vitro. Reduced levels of ninjurin1 impaired cardiac and skeletal muscle development in zebrafish. We conclude that Ninjurin1 contributes to myocyte growth and differentiation, and that these effects are mainly mediated by N-glycosylated Ninjurin1-24.


Assuntos
Estenose da Valva Aórtica/genética , Cardiomegalia/genética , Moléculas de Adesão Celular Neuronais/genética , Músculo Estriado/crescimento & desenvolvimento , Fatores de Crescimento Neural/genética , Animais , Estenose da Valva Aórtica/patologia , Cardiomegalia/patologia , Diferenciação Celular/genética , Modelos Animais de Doenças , Feminino , Humanos , Mutação com Perda de Função/genética , Masculino , Camundongos , Desenvolvimento Muscular/genética , Músculo Estriado/metabolismo , Músculo Estriado/patologia , Miócitos Cardíacos/metabolismo , Miócitos Cardíacos/patologia , Transdução de Sinais/genética , Peixe-Zebra
19.
Oncology ; 97(1): 26-37, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31071716

RESUMO

OBJECTIVE: Normally, activation of tropomyosin-related kinase (TRK) receptors by neurotrophins (NTs) stimulates intracellular pathways involved in cell survival and proliferation. Dysregulation of NT/TRK signaling may affect neoplasm prognosis. Data on NT and TRK expression in melanomas are limited, and it is unclear whether NT/TRK signaling pathways are involved in the origin and progression of this neoplasm. METHODS: We examined whether NT/TRK expression differs across different cutaneous melanoma grades and subtypes, and whether it is associated with melanoma prognosis and survival. A cross-sectional study was performed in which the expression of TrkA, TrkB, nerve growth factor (NGF), and brain-derived neurotrophic factor (BDNF) was analyzed by immunohistochemistry of 154 melanoma samples. We investigated NT/TRK expression associations with prognostic factors for melanoma, relapse-free survival (RFS), and overall survival (OS). RESULTS: Of the 154 melanoma samples, 77 (55.4%) were TrkA immunopositive, 81 (58.3%) were TrkB immunopositive, 113 (81.3%) were BDNF immunopositive, and 104 (75.4%) were NGF immunopositive. We found NT/TRK expression associated strongly with several clinical prognostic factors, including the tumor-node-metastasis stage (p < 0.001), histological subtype (p < 0.001), and Clark level (p < 0.05), as well as with a worse OS (p < 0.05 for all, except TrkB) and RFS (p < 0.05 for all). CONCLUSIONS: Our results show strong associations of NT/TRK expression with melanoma stage progression and a poor prognosis.


Assuntos
Fator Neurotrófico Derivado do Encéfalo/genética , Melanoma/genética , Glicoproteínas de Membrana/genética , Fatores de Crescimento Neural/genética , Receptor trkA/genética , Receptor trkB/genética , Neoplasias Cutâneas/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Intervalo Livre de Doença , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Melanoma/imunologia , Melanoma/patologia , Glicoproteínas de Membrana/imunologia , Pessoa de Meia-Idade , Fator de Crescimento Neural/genética , Fatores de Crescimento Neural/imunologia , Prognóstico , Receptor trkA/imunologia , Receptor trkB/imunologia , Transdução de Sinais , Neoplasias Cutâneas/imunologia , Neoplasias Cutâneas/patologia , Adulto Jovem
20.
Mol Med Rep ; 20(1): 813-829, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31115526

RESUMO

Infrapatellar fat pad­derived stem cells (IFPSCs) are emerging as an alternative to adipose tissue­derived stem cells (ADSCs) from other sources. They are a reliable source of autologous stem cells obtained from medical waste that are suitable for use in cell­based therapy, tissue engineering and regenerative medicine. Such clinical applications require a vast number of high­quality IFPSCs. Unlike embryonic stem cells (ESCs), ADSCs and IFPSCs have limited population doubling capacity; however, in vitro expansion of primary IFPSCs through multiple passages (referred to as P) is a crucial step to acquire the desired population of cells. The present study investigated the effect of multiple passages on the stemness of IFPSCs during expansion and the possibility of predicting the loss of stemness using certain markers. IFPSCs were isolated from infrapatellar fat pad tissue resected during knee arthroplasty performed on aged patients (>65 years old). These cells from the stromal vascular fraction were serially passaged to at least to P7, and their stemness characteristics were examined at each passage. It was observed that IFPSCs maintained their spindle­shaped morphology, self­renewability and homogeneity at P2­4. Furthermore, immunostaining revealed that these cells expressed mesenchymal stem cell (CD166, CD90 and CD105) and ESC markers [Sox2, Nanog, Oct4 and nucleostemin (NS)], whereas the hematopoietic stem cell marker CD45 was absent. These cells were also able to differentiate into the three germ layer cell types, thus confirming their ability to generate clinical grade cells. The findings indicated that prolonged culture of IFPSCs (P>6) led to the loss of the stem cell proliferative marker NS, with an increased population doubling time and progression toward neuronal differentiation, acquiring a neurogenic phenotype. Additionally, IFPSCs demonstrated an inherent ability to secrete neurotrophic factors and express receptors for these factors, which is the cause of neuronal differentiation at later passages. Therefore, these findings validated NS as a prognostic indicator for impaired stemness and identified IFPSCs as a promising source for cell­based therapy, particularly for neurodegenerative diseases.


Assuntos
Biomarcadores , Autorrenovação Celular/genética , Proteínas de Ligação ao GTP/genética , Células-Tronco Mesenquimais/citologia , Proteínas Nucleares/genética , Tecido Adiposo/citologia , Tecido Adiposo/metabolismo , Idoso , Diferenciação Celular/genética , Proliferação de Células/genética , Regulação da Expressão Gênica no Desenvolvimento/genética , Humanos , Células-Tronco Mesenquimais/metabolismo , Fatores de Crescimento Neural/genética , Doenças Neurodegenerativas/genética , Doenças Neurodegenerativas/patologia , Patela/citologia , Patela/metabolismo , Prognóstico
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