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1.
Am J Physiol Gastrointest Liver Physiol ; 318(4): G694-G704, 2020 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-32116022

RESUMO

Alcoholic pancreatitis is a multifactorial, progressive, inflammatory disorder of the pancreas. Alcohol initiates pancreatitis and promotes its progression in the context of genetic susceptibility and/or other environmental risk factors such as smoking. Genetic mutations can cause digestive enzyme misfolding, which induces endoplasmic reticulum (ER) stress and elicits pancreatitis. Here, we tested the hypothesis that alcohol synergizes with misfolding in promoting ER stress and thereby accelerates chronic pancreatitis progression. To this end, we fed an ethanol-containing diet to CPA1 N256K mice, which carry the human p.N256K CPA1 mutation and develop spontaneous chronic pancreatitis. Inexplicably, CPA1 N256K mice suffered generalized seizures after 2-3 wk of ethanol feeding, which resulted in high mortality and the early termination of the study. Analysis of CPA1 N256K mice euthanized after 3-3.5 wk of ethanol feeding revealed more severe chronic pancreatitis associated with significantly increased Hspa5 [ER chaperone immunoglobulin heavy chain-binding protein (BiP)] mRNA levels when compared with CPA1 N256K mice on a control liquid diet. In contrast, ethanol feeding of C57BL/6N mice for 4 wk increased Hspa5 levels to a lesser degree and caused no pancreatitis. We conclude that ethanol feeding synergizes with the misfolding CPA1 mutant in promoting ER stress and thereby accelerates progression of chronic pancreatitis in CPA1 N256K mice.NEW & NOTEWORTHY Alcoholic pancreatitis is a multifactorial, progressive, inflammatory disorder of the pancreas. This study demonstrates that alcohol synergizes with digestive enzyme misfolding in promoting endoplasmic reticulum stress and thereby accelerates progression of chronic pancreatitis.


Assuntos
Carboxipeptidases A/metabolismo , Etanol/toxicidade , Pâncreas/efeitos dos fármacos , Pancreatite Alcoólica/genética , Animais , Peso Corporal , Carboxipeptidases A/genética , Ingestão de Alimentos , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Etanol/administração & dosagem , Predisposição Genética para Doença , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Pancreatite Alcoólica/patologia
2.
Curr Med Sci ; 39(5): 727-733, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31612389

RESUMO

Hepatocellular carcinoma (HCC) has a poor treatment prognosis and high mortality worldwide. Understanding the molecular mechanism underlying HCC development would benefit the identification of diagnostic biomarkers and the improvement of the treatment strategies. The expression of carboxypeptidase A6 (CPA6) has been reported in epilepsy and febrile seizures rather than in any type of cancers. However, the function of CPA6 expression in HCC is not yet understood. In this study, we aimed to investigate the clinicopathological significance of the expression of CPA6 in HCC and the underlying mechanisms. We observed that the expression of the CPA6 protein was increased significantly in HCC tissues than in paracancerous tissues. To explore its function in HCC, both gain- and loss-of-function studies demonstrated that CPA6 played a vital role in promoting HCC growth and metastasis. When knocking down CPA6 with shRNA, HCC cell proliferation and migration could be suppressed. Meanwhile, CPA6 overexpression could promote proliferation and migration of HLF cells. Moreover, CPA6 could activate AKT serine/threonine kinase (AKT) signaling pathway as confirmed by Western blotting. In conclusion, our study revealed that CPA6 could promote HCC cell proliferation and migration via AKT-mediated signaling pathway. These findings suggest that CPA6 is a promising diagnostic biomarker and therapeutic target to improve the prognosis of HCC.


Assuntos
Carboxipeptidases A/genética , Carcinoma Hepatocelular/genética , Regulação Neoplásica da Expressão Gênica , Neoplasias Hepáticas/genética , Proteínas Proto-Oncogênicas c-akt/genética , Animais , Carboxipeptidases A/antagonistas & inibidores , Carboxipeptidases A/metabolismo , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Humanos , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia , Metástase Linfática , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Prognóstico , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Transdução de Sinais , Carga Tumoral , Ensaios Antitumorais Modelo de Xenoenxerto
3.
Mol Carcinog ; 58(11): 2026-2039, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31397502

RESUMO

Carboxypeptidase A4 (CPA4) is a member of the metallocarboxypeptidase family. A previous study indicated that CPA4 may participate in the modulation of peptide hormone activity and hormone-regulated tissue growth and differentiation. However, the role of CPA4 in lung tumorigenesis remains unclear. Our study revealed that CPA4 expression was higher in both lung cancer cells and tumor tissues. We performed 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assays, colony-formation assays, and Cellomics ArrayScan Infinity analysis to demonstrate that CPA4 knockdown inhibited non small-cell lung cancer (NSCLC) cell proliferation. Conversely, ectopic expression of CPA4 enhanced lung cancer cell proliferation. Consistent with these observations, we generated xenograft tumor models to confirm that CPA4 downregulation suppressed NSCLC cell growth. Mechanistically, we revealed that CPA4 downregulation may induce apoptosis and G1-S arrest by suppressing the protein kinase B/c-MYC pathway. These results suggest that CPA4 has an oncogenic effect on lung cancer growth. Taken together, we identified a novel gene in lung cancer that might provide a basis for new therapeutic targets.


Assuntos
Carboxipeptidases A/genética , Carcinoma Pulmonar de Células não Pequenas/genética , Proteína Oncogênica v-akt/genética , Proteínas Proto-Oncogênicas c-myc/genética , Animais , Apoptose/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Linhagem Celular Tumoral , Proliferação de Células/genética , Regulação Neoplásica da Expressão Gênica , Xenoenxertos , Humanos , Camundongos , Transdução de Sinais/genética
4.
Int J Biol Macromol ; 138: 125-134, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31279884

RESUMO

Carboxypeptidase A4 (CPA4) is a novel cancer-related gene that is aberrantly expressed in various malignant tumors. However, the roles and mechanisms of CPA4 have not been explored in colorectal cancer (CRC). In this study, we investigated the functions and mechanisms by which CPA4 promotes CRC progression. Quantitative real-time PCR (qRT-PCR) and western blot showed that CPA4 mRNA and CPA4 protein levels were up-regulated in CRC compared to levels in adjacent normal tissue. Immunohistochemistry (IHC) results indicating high CPA4 levels were positively associated with poor prognoses. In addition, Cell Counting Kit-8 (CCK-8), colony formation, flow cytometry, and transwell assays demonstrated that CPA4 overexpression facilitated the growth of CRC cells, whereas CPA4 knockdown resulted in decreased proliferation, G1/S phase transition arrest, and apoptosis. Subcutaneous tumorigenesis was performed in nude mice to confirm the tumor-promoting effects of CPA4 in vivo. Western blot revealed that activation of the STAT3 and ERK pathways is one of the oncogenic functions of CPA4 in CRC. Accordingly, CPA4 promotes CRC cell growth via activating the STAT3 and ERK pathways and may be a prognostic factor or therapeutic target for CRC.


Assuntos
Carboxipeptidases A/metabolismo , Neoplasias Colorretais/diagnóstico , Neoplasias Colorretais/patologia , Sistema de Sinalização das MAP Quinases , Fator de Transcrição STAT3/metabolismo , Idoso , Animais , Apoptose , Carboxipeptidases A/deficiência , Carboxipeptidases A/genética , Carcinogênese , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Transformação Celular Neoplásica , Neoplasias Colorretais/enzimologia , Feminino , Pontos de Checagem da Fase G1 do Ciclo Celular , Técnicas de Silenciamento de Genes , Humanos , Masculino , Camundongos , Pessoa de Meia-Idade
5.
Int J Exp Pathol ; 100(2): 133-138, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-31058377

RESUMO

Carboxypeptidase A4 (CPA4), a member of the metallo-carboxypeptidase family, is overexpressed in liver cancer and is associated with cancer progression. The role of CPA4 in hepatocellular carcinoma (HCC) remains unclear. In this study, we aimed to evaluate the relevance of CPA4 to the proliferation and expression of stem cell characteristics of hepatocellular carcinoma cells. Western blot analysis showed high CPA4 expression in the liver cancer cell line Bel7402 and low expression in HepG2 cells. Knock-down of CPA4 decreased cancer cell proliferation as detected by MTT and clone formation assays. The serum-free culture system revealed that downregulated CPA4 suppressed the sphere formation capacities of tumour cells. However, upregulated CPA4 increased the proliferation and sphere formation capacity. In addition, the protein expression of CD133, ALDH1 and CD44 also increased in cells with upregulated CPA4. In vivo, the overexpression of CPA4 in tumour cells that were subcutaneously injected into nude mice markedly increased the growth of the tumours. These data suggest that CPA4 expression leads to poor prognoses by regulating tumour proliferation and the expression of stem cell characteristics and may therefore serve as a potential therapeutic target of HCC.


Assuntos
Carboxipeptidases A/fisiologia , Carcinoma Hepatocelular/patologia , Neoplasias Hepáticas/patologia , Células-Tronco Neoplásicas/fisiologia , Animais , Carboxipeptidases A/deficiência , Carboxipeptidases A/genética , Carcinoma Hepatocelular/enzimologia , Linhagem Celular Tumoral , Proliferação de Células/fisiologia , Regulação para Baixo , Técnicas de Silenciamento de Genes/métodos , Células Hep G2 , Xenoenxertos , Humanos , Neoplasias Hepáticas/enzimologia , Masculino , Camundongos Nus , Transplante de Neoplasias , Células-Tronco Neoplásicas/patologia , Regulação para Cima
6.
Biomolecules ; 9(3)2019 03 14.
Artigo em Inglês | MEDLINE | ID: mdl-30875843

RESUMO

Recent research focused on prolonged survival has suggested that carboxypeptidase A4 (CPA4) plays a role in both tumor microenvironment formation and distant metastasis in cancer. In some patients, serum and expression (mRNA) levels of CPA4 have been found to be correlated with the aggressiveness and progression of the disease. Accordingly, we conducted a first study to investigate the diagnostic and prognostic significance of CPA4 in the case of breast cancer (BC), the most common form of malignancy in women. The study included a total of 50 patients with BC and 20 healthy women as the control group. The participants' serum CPA4 levels were determined by the ELISA test, and, for assessment of CPA4 mRNA, we used the PCR method. The serum CPA4 (p = 0.001) and CPA4 mRNA (p = 0.015) levels were found to be statistically significantly higher in the controls, compared to the patient group. When the results of patient group were statistically analyzed based on subgrouping by tumor characteristics, the measured CPA4 mRNA levels showed significant difference with respect to the molecular subtype (p = 0.006), pN status (p = 0.023), and pathological stage (p = 0.039), while the serum CPA4 measurements differed significantly in terms of pathological type only (p = 0.024). We conclude that CPA4 is diagnostically and prognostically not futile when used in combination with the other considerations and measurements in breast cancer.


Assuntos
Biomarcadores Tumorais/sangue , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/enzimologia , Carboxipeptidases A/sangue , Neoplasias da Mama/sangue , Neoplasias da Mama/metabolismo , Carboxipeptidases A/genética , Carboxipeptidases A/metabolismo , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Pessoa de Meia-Idade , RNA Neoplásico/sangue , RNA Neoplásico/genética , RNA Neoplásico/isolamento & purificação
7.
Int J Oncol ; 54(3): 833-844, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30628666

RESUMO

Using whole transcriptome analysis and a lentiviral short hairpin RNA screening library, carboxypeptidase A4 (CPA4) was identified as a novel marker in breast cancer and a therapeutic target in triple­negative breast cancer (TNBC) in the present study. Immunohistochemistry was used to evaluate the presence of CPA4, estrogen receptor, progesterone receptor, human epidermal growth factor receptor 2, Ki67, epidermal growth factor receptor, cytokeratin 5/6, aldehyde dehydrogenase 1, cluster of differentiation (CD)44, CD24, claudins, E­cadherin, vimentin and androgen receptor in 221 cases of breast cancer, including 68 TNBC cases. The effects of CPA4 on the viability and migration ability of TNBC cells were analyzed using RNA interference methods. Increased CPA4 expression, specifically in the cytoplasm of cancer tissue cells, was detected. Furthermore, high CPA4 expression in TNBC cases was associated with low expression of E­cadherin and with the expression of cancer stem cell markers (high CD44/low CD24). Patients with TNBC and high levels of CPA4 expression had a significantly poorer prognosis compared with those with low CPA4 expression. Notably, viability and migration were reduced, but E­cadherin expression was upregulated in CPA4­suppressed TNBC cells. The present data suggested that CPA4 may be a novel inducer for epithelial­mesenchymal transition, which is characterized by the downregulation of E­cadherin and mesenchymal phenotypes. To conclude, CPA4 may be a marker for poor prognosis and a promising therapeutic target in TNBC with aggressive phenotypes.


Assuntos
Carboxipeptidases A/metabolismo , Neoplasias de Mama Triplo Negativas/diagnóstico , Neoplasias de Mama Triplo Negativas/patologia , Adulto , Idoso , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Carboxipeptidases A/genética , Linhagem Celular Tumoral , Movimento Celular , Sobrevivência Celular , Citoplasma/metabolismo , Feminino , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Humanos , Pessoa de Meia-Idade , Fenótipo , Prognóstico , RNA Interferente Pequeno/metabolismo , Análise de Sobrevida , Neoplasias de Mama Triplo Negativas/genética
8.
J Med Chem ; 62(4): 1917-1931, 2019 02 28.
Artigo em Inglês | MEDLINE | ID: mdl-30688452

RESUMO

Metallocarboxypeptidases (MCPs) of the M14 family are Zn2+-dependent exoproteases present in almost every tissue or fluid in mammals. These enzymes perform a large variety of physiological functions and are involved in several pathologies, such as pancreatic diseases, inflammation, fibrinolysis, and cancer. Here, we describe the synthesis and functional/structural characterization of a series of reversible tight-binding phosphinic pseudopeptide inhibitors that show high specificity and potency toward these proteases. Characterization of their inhibitory potential against a large variety of MCPs, combined with high-resolution crystal structures of three selected candidates in complex with human carboxypeptidase A (CPA)1, allowed to decipher the structural determinants governing selectivity for type-A of the M14A MCP family. Further, the phosphinic pseudopeptide framework was exploited to generate an optical probe selectively targeting human CPAs. The phosphinic pseudopeptides presented here constitute the first example of chemical probes useful to selectively report on type-A MCPs activity in complex media.


Assuntos
Carboxipeptidases A/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Oligopeptídeos/farmacologia , Ácidos Fosfínicos/farmacologia , Carboxipeptidases A/química , Carboxipeptidases A/metabolismo , Domínio Catalítico , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/metabolismo , Corantes Fluorescentes/síntese química , Corantes Fluorescentes/farmacologia , Células HEK293 , Células HeLa , Humanos , Indóis/síntese química , Indóis/farmacologia , Cinética , Microscopia Confocal/métodos , Microscopia de Fluorescência/métodos , Oligopeptídeos/síntese química , Oligopeptídeos/metabolismo , Ácidos Fosfínicos/síntese química , Ácidos Fosfínicos/metabolismo , Ligação Proteica
9.
Gut ; 68(2): 301-312, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30045879

RESUMO

OBJECTIVE: Chronic pancreatitis is a progressive, relapsing inflammatory disorder of the pancreas, which often develops in the background of genetic susceptibility. Recently, loss-of-function mutations in CPA1, which encodes the digestive enzyme carboxypeptidase A1, were described in sporadic early onset cases and in hereditary pancreatitis. Mutation-induced misfolding of CPA1 and associated endoplasmic reticulum (ER) stress was suggested as potential disease mechanism; however, in vivo evidence has been lacking. The objective of the present study was to create a mouse model that recapitulates features of CPA1-associated chronic pancreatitis. DESIGN: We knocked-in the most frequently occurring p.N256K human CPA1 mutation to the mouse Cpa1 locus. Mutant mice were characterised with respect to pancreas pathology and ER stress and compared with C57BL/6N and CPA1 null control mice. RESULTS: In the CPA1 N256K mutant mice, we observed hallmarks of chronic pancreatitis that included progressive acinar cell atrophy, inflammatory cell infiltration, fibrosis and acinar-ductal metaplasia. In contrast, similarly to the C57BL/6N mice, the CPA1 null control strain exhibited no signs of pancreatic disease. Mutation p.N256K induced misfolding of mouse CPA1 and resulted in elevated expression of ER stress markers Hspa5 (BiP) and Ddit3 (CHOP) both in cell culture and mutant mice. CONCLUSION: The results offer categorical evidence that CPA1 mutations elicit enzyme misfolding and cause chronic pancreatitis via an ER stress-related mechanism.


Assuntos
Carboxipeptidases A/genética , Pancreatite Crônica/enzimologia , Pancreatite Crônica/genética , Dobramento de Proteína , Animais , Modelos Animais de Doenças , Progressão da Doença , Estresse do Retículo Endoplasmático/genética , Predisposição Genética para Doença , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Mutação
10.
Zh Nevrol Psikhiatr Im S S Korsakova ; 119(11. Vyp. 2): 65-69, 2019.
Artigo em Russo | MEDLINE | ID: mdl-32207733

RESUMO

A clinical case of familial temporal epilepsy type 5 (OMIM 614417) with onset at the age of 14 years is described for the first time in the domestic literature. The leading manifestations of the disease were focal seizures of systemic vertigo, accompanied by vestibular ataxia and, sometimes, vomiting. Cognitive and emotional disturbances were observed as well. On the EEG of wakefulness and sleep, a non-expressed epileptiform activity was detected in the left parietotemporal and frontotemporal zone. Neuroimaging did not show any significant changes. Only a molecular genetic study that identified CPA6 gene mutation made it possible to establish the accurate diagnosis.


Assuntos
Epilepsia do Lobo Temporal , Adolescente , Carboxipeptidases A/genética , Eletroencefalografia , Epilepsia do Lobo Temporal/diagnóstico , Epilepsia do Lobo Temporal/genética , Epilepsia do Lobo Temporal/fisiopatologia , Epilepsia do Lobo Temporal/psicologia , Humanos , Mutação , Convulsões/complicações , Vigília
11.
Artigo em Inglês | MEDLINE | ID: mdl-30275084

RESUMO

The Gram-negative human pathogen Neisseria gonorrhoeae has progressively developed resistance to antibiotic monotherapies, and recent failures of dual-drug therapy have heightened concerns that strains resistant to all available antibiotics will begin circulating globally. Targeting bacterial cell wall assembly has historically been effective at treating infections with N. gonorrhoeae, but as the effectiveness of ß-lactams (including cephalosporins) is challenged by increasing resistance, research has expanded into compounds that target the numerous other enzymes with roles in peptidoglycan metabolism. One example is the dithiazoline compound JNJ-853346 (DTZ), which inhibits the activity of an Escherichia coli serine protease l,d-carboxypeptidase (LdcA). Recently, the characterization of an LdcA homolog in N. gonorrhoeae revealed localization and activity differences from the characterized E. coli LdcA, prompting us to explore the effectiveness of DTZ against N. gonorrhoeae We found that DTZ is effective at inhibiting N. gonorrhoeae in all growth phases, unlike the specific stationary-phase inhibition seen in E. coli Surprisingly, DTZ does not inhibit gonococcal LdcA enzyme activity, and DTZ sensitivity is not significantly decreased in ldcA mutants. While effective against numerous N. gonorrhoeae strains, including recent multidrug-resistant isolates, DTZ is much less effective at inhibiting growth of the commensal species Lactobacillus gasseri DTZ treatment during coinfections of epithelial cells resulted in significant lowering of gonococcal burden and interleukin-8 secretion without significantly impacting recovery of viable L. gasseri This selective toxicity presents a possible pathway for the use of DTZ as an effective antigonococcal agent at concentrations that do not impact vaginal commensals.


Assuntos
Antibacterianos/farmacologia , Parede Celular/efeitos dos fármacos , Lactobacillus gasseri/efeitos dos fármacos , Neisseria gonorrhoeae/efeitos dos fármacos , Tiazóis/farmacologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Carboxipeptidases A/genética , Carboxipeptidases A/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Parede Celular/metabolismo , Expressão Gênica , Células HCT116 , Humanos , Interleucina-8/genética , Interleucina-8/imunologia , Lactobacillus gasseri/crescimento & desenvolvimento , Lactobacillus gasseri/metabolismo , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacos , Mutação , Neisseria gonorrhoeae/genética , Neisseria gonorrhoeae/crescimento & desenvolvimento , Neisseria gonorrhoeae/metabolismo , Peptidoglicano/biossíntese , Peptidoglicano/efeitos dos fármacos , Probióticos/química , Especificidade da Espécie
12.
Gastrointest Endosc Clin N Am ; 28(4): 587-603, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30241646

RESUMO

Individuals with acute recurrent and chronic pancreatitis may have an inherited predisposition to the development of the disease. Pancreatitis in the setting of a significant family history of the disease can be classified as hereditary or familial pancreatitis. In this article, the authors closely examine the specific genes implicated in pancreatitis, investigate the role of genetic testing for diagnosis, and describe the impact of genetic testing results on clinical management.


Assuntos
Regulador de Condutância Transmembrana em Fibrose Cística/genética , Predisposição Genética para Doença , Neoplasias Pancreáticas/genética , Pancreatite/genética , Inibidor da Tripsina Pancreática de Kazal/genética , Tripsina/genética , Carboxipeptidases A/genética , Quimotripsina/genética , Claudina-2/genética , Testes Genéticos , Mutação em Linhagem Germinativa , Humanos , Pancreatite/terapia , Receptores de Detecção de Cálcio/genética , Medição de Risco
13.
J Neuroinflammation ; 15(1): 265, 2018 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-30217166

RESUMO

BACKGROUND: It remains unclear whether disease course in multiple sclerosis (MS) is influenced by genetic polymorphisms. Here, we aimed to identify genetic variants associated with benign and aggressive disease courses in MS patients. METHODS: MS patients were classified into benign and aggressive phenotypes according to clinical criteria. We performed exome sequencing in a discovery cohort, which included 20 MS patients, 10 with benign and 10 with aggressive disease course, and genotyping in 2 independent validation cohorts. The first validation cohort encompassed 194 MS patients, 107 with benign and 87 with aggressive phenotypes. The second validation cohort comprised 257 patients, of whom 224 patients had benign phenotypes and 33 aggressive disease courses. Brain immunohistochemistries were performed using disease course associated genes antibodies. RESULTS: By means of single-nucleotide polymorphism (SNP) detection and comparison of allele frequencies between patients with benign and aggressive phenotypes, a total of 16 SNPs were selected for validation from the exome sequencing data in the discovery cohort. Meta-analysis of genotyping results in two validation cohorts revealed two polymorphisms, rs28469012 and rs10894768, significantly associated with disease course. SNP rs28469012 is located in CPXM2 (carboxypeptidase X, M14 family, member 2) and was associated with aggressive disease course (uncorrected p value < 0.05). SNP rs10894768, which is positioned in IGSF9B (immunoglobulin superfamily member 9B) was associated with benign phenotype (uncorrected p value < 0.05). In addition, a trend for association with benign phenotype was observed for a third SNP, rs10423927, in NLRP9 (NLR family pyrin domain containing 9). Brain immunohistochemistries in chronic active lesions from MS patients revealed expression of IGSF9B in astrocytes and macrophages/microglial cells, and expression of CPXM2 and NLRP9 restricted to brain macrophages/microglia. CONCLUSIONS: Genetic variants located in CPXM2, IGSF9B, and NLRP9 have the potential to modulate disease course in MS patients and may be used as disease activity biomarkers to identify patients with divergent disease courses. Altogether, the reported results from this study support the influence of genetic factors in MS disease course and may help to better understand the complex molecular mechanisms underlying disease pathogenesis.


Assuntos
Predisposição Genética para Doença/genética , Esclerose Múltipla/genética , Esclerose Múltipla/fisiopatologia , Polimorfismo de Nucleotídeo Único/genética , Sequenciamento Completo do Exoma/métodos , Encéfalo/metabolismo , Carboxipeptidases A/genética , Carboxipeptidases A/metabolismo , Estudos de Coortes , Progressão da Doença , Feminino , Frequência do Gene , Genótipo , Humanos , Imunoglobulinas/genética , Imunoglobulinas/metabolismo , Masculino , Esclerose Múltipla/patologia , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , RNA Mensageiro
14.
J Biomed Inform ; 84: 159-163, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-30004020

RESUMO

In the past, algorithms exploiting varying semantics in interactions between biological objects such as genes and diseases have been used in bioinformatics to uncover latent relationships within biological datasets. In this paper, we consider the algorithm Medusa in parallel with binary classification in order to find potential compounds to inhibit oral cancer. Oral cancer affects the mouth and pharynx and has a high mortality rate due to its late discovery. Current methods of oral cancer treatment, such as chemoradiation and surgery, fail to provide better chances for survival, warranting an alternative approach. By running Medusa on a data fusion graph consisting of biological objects, we incorporate binary classification to model the algorithm's association detection to discover compounds with the potential to mitigate the effects of oral cancer.


Assuntos
Antineoplásicos/farmacologia , Biologia Computacional , Neoplasias Bucais/tratamento farmacológico , Preparações Farmacêuticas/química , Algoritmos , Carboxipeptidases A/antagonistas & inibidores , Química Farmacêutica , Simulação por Computador , Quinase 2 Dependente de Ciclina/antagonistas & inibidores , Bases de Dados Genéticas , Desenho de Fármacos , Humanos , Mutação , Poli(ADP-Ribose) Polimerase-1/antagonistas & inibidores , Reprodutibilidade dos Testes , Semântica
15.
Curr Opin Allergy Clin Immunol ; 18(5): 370-376, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-30048251

RESUMO

PURPOSE OF REVIEW: The aim of the review is to describe the different clinical pictures of anaphylaxis (phenotypes), in relation to the underlying mechanisms and potential biomarkers, to describe anaphylaxis endotypes. This may aid in achieving a better understanding, management and outcomes of such severe reactions. RECENT FINDINGS: Different anaphylaxis phenotypes have been outlined, ranging from the classical type-I-like to those suggestive of cytokine-storm-like or complement-mediated reactions. Underlying mechanisms differ and biomarkers of cells and systems involved are being identified (tryptase, IL-6, bradykinin etc.) SUMMARY: Identifying specific phenotypes/endotypes will allow the application of precision medicine in patients with anaphylaxis, providing insights to the most appropriate approach in each case.


Assuntos
Anafilaxia/imunologia , Biomarcadores/metabolismo , Ativação do Complemento/imunologia , Citocinas/imunologia , Fenótipo , Anafilaxia/metabolismo , Basófilos/imunologia , Bradicinina/imunologia , Bradicinina/metabolismo , Carboxipeptidases A/imunologia , Carboxipeptidases A/metabolismo , Quimases/imunologia , Quimases/metabolismo , Citocinas/metabolismo , Histamina/imunologia , Histamina/metabolismo , Humanos , Imunoglobulina E/imunologia , Imunoglobulina G/imunologia , Interleucina-6/imunologia , Interleucina-6/metabolismo , Mastócitos/imunologia , Fator de Ativação de Plaquetas/imunologia , Fator de Ativação de Plaquetas/metabolismo , Medicina de Precisão , Triptases/imunologia , Triptases/metabolismo
16.
Proc Natl Acad Sci U S A ; 115(18): 4767-4772, 2018 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-29669919

RESUMO

To evaluate whether germline variants in genes encoding pancreatic secretory enzymes contribute to pancreatic cancer susceptibility, we sequenced the coding regions of CPB1 and other genes encoding pancreatic secretory enzymes and known pancreatitis susceptibility genes (PRSS1, CPA1, CTRC, and SPINK1) in a hospital series of pancreatic cancer cases and controls. Variants in CPB1, CPA1 (encoding carboxypeptidase B1 and A1), and CTRC were evaluated in a second set of cases with familial pancreatic cancer and controls. More deleterious CPB1 variants, defined as having impaired protein secretion and induction of endoplasmic reticulum (ER) stress in transfected HEK 293T cells, were found in the hospital series of pancreatic cancer cases (5/986, 0.5%) than in controls (0/1,045, P = 0.027). Among familial pancreatic cancer cases, ER stress-inducing CPB1 variants were found in 4 of 593 (0.67%) vs. 0 of 967 additional controls (P = 0.020), with a combined prevalence in pancreatic cancer cases of 9/1,579 vs. 0/2,012 controls (P < 0.01). More ER stress-inducing CPA1 variants were also found in the combined set of hospital and familial cases with pancreatic cancer than in controls [7/1,546 vs. 1/2,012; P = 0.025; odds ratio, 9.36 (95% CI, 1.15-76.02)]. Overall, 16 (1%) of 1,579 pancreatic cancer cases had an ER stress-inducing CPA1 or CPB1 variant, compared with 1 of 2,068 controls (P < 0.00001). No other candidate genes had statistically significant differences in variant prevalence between cases and controls. Our study indicates ER stress-inducing variants in CPB1 and CPA1 are associated with pancreatic cancer susceptibility and implicate ER stress in pancreatic acinar cells in pancreatic cancer development.


Assuntos
Carboxipeptidase B , Carboxipeptidases A , Estresse do Retículo Endoplasmático/genética , Predisposição Genética para Doença , Mutação , Proteínas de Neoplasias , Neoplasias Pancreáticas , Idoso , Idoso de 80 Anos ou mais , Carboxipeptidase B/genética , Carboxipeptidase B/metabolismo , Carboxipeptidases A/genética , Carboxipeptidases A/metabolismo , Linhagem Celular Tumoral , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Neoplasias Pancreáticas/enzimologia , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/patologia
17.
Diabetes ; 67(7): 1428-1440, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29650774

RESUMO

Metformin is the first-line treatment for type 2 diabetes (T2D). Although widely prescribed, the glucose-lowering mechanism for metformin is incompletely understood. Here, we used a genome-wide association approach in a diverse group of individuals with T2D from the Action to Control Cardiovascular Risk in Diabetes (ACCORD) clinical trial to identify common and rare variants associated with HbA1c response to metformin treatment and followed up these findings in four replication cohorts. Common variants in PRPF31 and CPA6 were associated with worse and better metformin response, respectively (P < 5 × 10-6), and meta-analysis in independent cohorts displayed similar associations with metformin response (P = 1.2 × 10-8 and P = 0.005, respectively). Previous studies have shown that PRPF31(+/-) knockout mice have increased total body fat (P = 1.78 × 10-6) and increased fasted circulating glucose (P = 5.73 × 10-6). Furthermore, rare variants in STAT3 associated with worse metformin response (q <0.1). STAT3 is a ubiquitously expressed pleiotropic transcriptional activator that participates in the regulation of metabolism and feeding behavior. Here, we provide novel evidence for associations of common and rare variants in PRPF31, CPA6, and STAT3 with metformin response that may provide insight into mechanisms important for metformin efficacy in T2D.


Assuntos
Carboxipeptidases A/genética , Diabetes Mellitus Tipo 2/tratamento farmacológico , Diabetes Mellitus Tipo 2/genética , Proteínas do Olho/genética , Metformina/uso terapêutico , Variantes Farmacogenômicos , Estudos de Coortes , Método Duplo-Cego , Feminino , Estudo de Associação Genômica Ampla , Humanos , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , Fator de Transcrição STAT3/genética , Resultado do Tratamento
18.
J Allergy Clin Immunol ; 142(2): 381-393, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29454835

RESUMO

FcεRI is the primary receptor in mast cells that mediates allergic reactions by inducing rapid release of mediators, an adaptive immune response that might have evolved as a host defense against parasites and venoms. Yet it is apparent that mast cells are also activated through non-IgE receptors, the significance of which is just beginning to be understood. This includes the Mas-related G protein-coupled receptor X2, which might contribute to reactions to diverse antimicrobials and polybasic compounds, and the adhesion G protein-coupled receptor E2, variants of which are associated with familial vibratory urticaria and are activated by mechanical vibration. Similarly, mast cells have long been recognized as the main repository for histamine, heparin, and proteases. Recent evidence also points to new functions, modes of delivery, and mechanisms of action of mast cell proteases that add new dimensions to the roles of mast cells in human biology. In addition, exposure of mast cells to environmental cues can quantitatively and qualitatively modulate their responses and thus their effect on allergic inflammation. Illustrating this paradigm, we summarize a number of recent studies implicating the injury/tissue damage cytokine IL-33 as a modulator of allergen-induced mast cell responses. We also discuss the discovery of markers associated with transformed mast cells and new potential directions in suppressing mast cell activity.


Assuntos
Hipersensibilidade/imunologia , Inflamação/imunologia , Interleucina-33/metabolismo , Mastócitos/imunologia , Mastocitose/imunologia , Receptores de IgE/metabolismo , Urticária/imunologia , Animais , Carboxipeptidases A/metabolismo , Degranulação Celular , Histamina/metabolismo , Humanos , Proteínas do Tecido Nervoso/metabolismo , Receptores Acoplados a Proteínas-G/metabolismo , Receptores de Neuropeptídeos/metabolismo , Transdução de Sinais , Vibração
19.
Appl Biochem Biotechnol ; 185(4): 1029-1043, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29404908

RESUMO

Carboxypeptidase A (CPA) is a metalloexopeptidase that catalyzes the hydrolysis of the peptide bonds that are adjacent to the C-terminal end of a polypeptide chain. The enzyme preferentially cleaves over C-terminal L-amino acids with aromatic or branched side chains. This is of main importance for food industry because it can be employed for manufacturing functional foods from different protein sources with reduced hydrophobic amino acid content for patients with deficiencies in the absorption or digestion of the corresponding amino acids. In that way, strategies for effective multipoint covalent immobilization of CPA metalloenzyme on chitosan beads have been developed. The study of the ability to produce several chemical modifications on chitosan molecules before, during and after its coagulation to form carrier beads lead in a protective effect of the polymer matrix. The chemical modification of chitosan through the use of an N-alkylation strategy produced the best derivatives. N-alkyl chitosan derivative beads with D-fructose presented values of 0.86 for immobilization yield, 314.6 IU g-1 bead for initial activity of biocatalyst and were 5675.64-fold more stable than the free enzyme at 55 °C. Results have shown that these derivatives would present a potential technological application in hydrolytic processes due to both their physical properties, such as low swelling capacity, reduced metal chelation ability and bulk mesoporosity, and increased operational stability when compared with soluble enzyme.


Assuntos
Carboxipeptidases A/química , Quitosana/química , Enzimas Imobilizadas/química , Biocatálise , Estabilidade Enzimática , Frutose/química , Temperatura Alta
20.
Front Immunol ; 9: 207, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29479356

RESUMO

Infection with the helminth parasite Strongyloides stercoralis (Ss) is commonly clinically asymptomatic that is often accompanied by peripheral eosinophilia. Granulocytes are activated during helminth infection and can act as immune effector cells. Plasma levels of eosinophil and neutrophil granular proteins convey an indirect measure of granulocyte degranulation and are prominently augmented in numerous helminth-infected patients. In this study, we sought to examine the levels of eosinophil, neutrophil, and mast cell activation-associated granule proteins in asymptomatic Ss infection and to understand their kinetics following anthelmintic therapy. To this end, we measured the plasma levels of eosinophil cationic protein, eosinophil-derived neurotoxin, eosinophil peroxidase, eosinophil major basic protein, neutrophil elastase, myeloperoxidase, neutrophil proteinase-3, mast cell tryptase, leukotriene C4, and mast cell carboxypeptidase-A3 in individuals with asymptomatic Ss infection or without Ss infection [uninfected (UN)]. We also estimated the levels of all of these analytes in infected individuals following definitive treatment of Ss infection. We demonstrated that those infected individuals have significantly enhanced plasma levels of eosinophil cationic protein, eosinophil-derived neurotoxin, eosinophil peroxidase, eosinophil major basic protein, elastase, myeloperoxidase, mast cell tryptase, leukotriene C4, and carboxypeptidase-A3 compared to UN individuals. Following the treatment of Ss infection, each of these granulocyte-associated proteins drops significantly. Our data suggest that eosinophil, neutrophil, and mast cell activation may play a role in the response to Ss infection.


Assuntos
Proteínas Granulares de Eosinófilos/sangue , Eosinófilos/imunologia , Mastócitos/imunologia , Neutrófilos/imunologia , Strongyloides stercoralis/imunologia , Estrongiloidíase/sangue , Adulto , Animais , Antiprotozoários/uso terapêutico , Infecções Assintomáticas/terapia , Carboxipeptidases A/sangue , Carboxipeptidases A/imunologia , Carboxipeptidases A/metabolismo , Proteínas Granulares de Eosinófilos/imunologia , Proteínas Granulares de Eosinófilos/metabolismo , Eosinófilos/metabolismo , Feminino , Interações Hospedeiro-Parasita/imunologia , Humanos , Elastase de Leucócito/sangue , Elastase de Leucócito/imunologia , Elastase de Leucócito/metabolismo , Leucotrieno C4/sangue , Leucotrieno C4/imunologia , Leucotrieno C4/metabolismo , Masculino , Mastócitos/metabolismo , Pessoa de Meia-Idade , Neutrófilos/metabolismo , Peroxidase/sangue , Peroxidase/imunologia , Peroxidase/metabolismo , Vesículas Secretórias/imunologia , Vesículas Secretórias/metabolismo , Strongyloides stercoralis/isolamento & purificação , Estrongiloidíase/tratamento farmacológico , Estrongiloidíase/imunologia , Estrongiloidíase/parasitologia , Resultado do Tratamento , Triptases/sangue , Triptases/imunologia , Triptases/metabolismo , Adulto Jovem
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