Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 5 de 5
Filter
1.
Cell Biol Int ; 46(2): 255-264, 2022 Feb.
Article in English | MEDLINE | ID: mdl-34816536

ABSTRACT

Although the elevated level of the α-N-acetylgalactosaminidase enzyme (encoded by the NAGA gene) is a well-recognized feature of cancer cells; little research works have been undertaken on the cancer malignancy mechanisms. The effects of NAGA gene downregulation on cancer cells' features such as drug resistance, impaired programmed cell death, and migration were analyzed in this study. The cells grew exponentially with a doubling time of 30 h in an optimal condition. Toxicity of daunorubicin chemotherapy drug on NAGA-transfected EPG85.257RDB cells was evaluated in comparison to control cells and no significant change was recorded. Quantitative transcript analyses and protein levels revealed that the MDR1 pump almost remained unchanged during the study. Moreover, the NAGA gene downregulation enhanced the late apoptosis rate in EPG85.257RDB cells at 24 h posttransfection. The investigated expression level of genes and proteins involved in the TNFR2 signaling pathway, related to cancer cell apoptosis, showed considerable alterations after NAGA silencing as well. MAP3K14 and CASP3 genes were downregulated while IL6, RELA, and TRAF2 experienced an upregulation. Also, NAGA silencing generally diminished the migration ability of EPG85.257RDB cells and the MMP1 gene (as a critical gene in metastasis) expression decreased significantly. The expression of the p-FAK protein, which is located in the downstream of the α2 ß1 integrin signaling pathway, was reduced likewise. It could be concluded that despite drug resistance, NAGA silencing resulted in augmentative and regressive effects on cell death and migration.


Subject(s)
Stomach Neoplasms , Apoptosis , Cell Death , Cell Line, Tumor , Drug Resistance, Multiple , Humans , Stomach Neoplasms/metabolism , alpha-N-Acetylgalactosaminidase/genetics , alpha-N-Acetylgalactosaminidase/metabolism , alpha-N-Acetylgalactosaminidase/therapeutic use
2.
Int J Mol Sci ; 22(5)2021 Mar 06.
Article in English | MEDLINE | ID: mdl-33800950

ABSTRACT

Male patients with Fabry disease (FD) are at high risk for the formation of antibodies to recombinant α-galactosidase A (AGAL), used for enzyme replacement therapy. Due to the rapid disease progression, the identification of patients at risk is highly warranted. However, currently suitable references and standardized protocols for anti-drug antibodies (ADA) determination do not exist. Here we generate a comprehensive patient-derived antibody mixture as a reference, allowing ELISA-based quantification of antibody titers from individual blood samples. Serum samples of 22 male patients with FD and ADAs against AGAL were pooled and purified by immune adsorption. ADA-affinities against agalsidase-α, agalsidase-ß and Moss-AGAL were measured by quartz crystal microbalance with dissipation monitoring (QCM-D). AGAL-specific immune adsorption generated a polyclonal ADA mixture showing a concentration-dependent binding and inhibition of AGAL. Titers in raw sera and from purified total IgGs (r2 = 0.9063 and r2 = 0.8952, both p < 0.0001) correlated with the individual inhibitory capacities of ADAs. QCM-D measurements demonstrated comparable affinities of the reference antibody for agalsidase-α, agalsidase-ß and Moss-AGAL (KD: 1.94 ± 0.11 µM, 2.46 ± 0.21 µM, and 1.33 ± 0.09 µM, respectively). The reference antibody allows the ELISA-based ADA titer determination and quantification of absolute concentrations. Furthermore, ADAs from patients with FD have comparable affinities to agalsidase-α, agalsidase-ß and Moss-AGAL.


Subject(s)
Antibodies, Neutralizing/immunology , Enzyme Replacement Therapy , Enzyme-Linked Immunosorbent Assay , Fabry Disease/immunology , alpha-Galactosidase/immunology , alpha-N-Acetylgalactosaminidase/immunology , Antibodies, Neutralizing/biosynthesis , Antibody Affinity , Dose-Response Relationship, Immunologic , Fabry Disease/blood , Fabry Disease/drug therapy , Humans , Immunoglobulin G/biosynthesis , Immunoglobulin G/immunology , Male , Recombinant Proteins/immunology , Recombinant Proteins/therapeutic use , Reference Standards , alpha-Galactosidase/blood , alpha-Galactosidase/therapeutic use , alpha-N-Acetylgalactosaminidase/blood , alpha-N-Acetylgalactosaminidase/therapeutic use
3.
J BUON ; 22(6): 1372-1377, 2017.
Article in English | MEDLINE | ID: mdl-29332325

ABSTRACT

In the constant battle against cancer cells, macrophages are of great importance. Their activation is achieved through various mechanisms such as Vitamin D binding protein (VDBP or Gc). After undergoing modifications via enzymes secreted by stimulated lymphocytes, VDBP is modified into Macrophages Activator Form/Factor (Gc-MAF). Some studies (particularly those focusing on cancer) have reported that an enzyme known as α-N-acetylgalactosaminidase (nagalase) facilitates the deglycosylation of Gc-MAF, which in turn inhibits the activation of macrophages. The aim of this review was to evaluate studies associated with nagalase and its escalation in various diseases and to propose hypothetical solutions in order to neutralize the effects of nagalase in cancer patients.


Subject(s)
Macrophage-Activating Factors/therapeutic use , Macrophages/metabolism , Neoplasms/drug therapy , Vitamin D-Binding Protein/therapeutic use , alpha-N-Acetylgalactosaminidase/therapeutic use , Humans , Macrophage-Activating Factors/pharmacology , Neoplasms/pathology , Vitamin D-Binding Protein/pharmacology , alpha-N-Acetylgalactosaminidase/pharmacology
4.
Rev Invest Clin ; 63(3): 314-21, 2011.
Article in Spanish | MEDLINE | ID: mdl-21888295

ABSTRACT

Fabry-Anderson disease is a lysosomal storage disease caused by deficiency of the enzyme alpha-galactosidase. This enzymatic defect results in the accumulation of glycosphingolipid into different lines cells. Usually the deficiency is complete, resulting in a multisystem disorder, with injury in different organs, predominantly heart, kidney and nervous system. However, in some patients the enzymatic deficit is partial and causes diverse clinical variants of the disease (renal or cardiac variety), this cause a difficult diagnostic and the absence of real epidemiology data. This review is about the epidemiology, the metabolic defect of this disease, it's molecular and genetics bases, the different forms of clinical presentation and the enzyme replacement therapy.


Subject(s)
Fabry Disease , Chromosomes, Human, X/genetics , Cohort Studies , Endothelium, Vascular/enzymology , Enzyme Replacement Therapy , Fabry Disease/diagnosis , Fabry Disease/drug therapy , Fabry Disease/enzymology , Fabry Disease/epidemiology , Fabry Disease/genetics , Humans , Kidney/enzymology , Lysosomes/enzymology , Male , Myocardium/enzymology , Organ Specificity , Phenotype , Randomized Controlled Trials as Topic , alpha-Galactosidase/analysis , alpha-Galactosidase/biosynthesis , alpha-Galactosidase/genetics , alpha-Galactosidase/therapeutic use , alpha-N-Acetylgalactosaminidase/therapeutic use
5.
Am J Hum Genet ; 85(5): 569-80, 2009 Nov.
Article in English | MEDLINE | ID: mdl-19853240

ABSTRACT

A modified alpha-N-acetylgalactosaminidase (NAGA) with alpha-galactosidase A (GLA)-like substrate specificity was designed on the basis of structural studies and was produced in Chinese hamster ovary cells. The enzyme acquired the ability to catalyze the degradation of 4-methylumbelliferyl-alpha-D-galactopyranoside. It retained the original NAGA's stability in plasma and N-glycans containing many mannose 6-phosphate (M6P) residues, which are advantageous for uptake by cells via M6P receptors. There was no immunological cross-reactivity between the modified NAGA and GLA, and the modified NAGA did not react to serum from a patient with Fabry disease recurrently treated with a recombinant GLA. The enzyme cleaved globotriaosylceramide (Gb3) accumulated in cultured fibroblasts from a patient with Fabry disease. Furthermore, like recombinant GLA proteins presently used for enzyme replacement therapy (ERT) for Fabry disease, the enzyme intravenously injected into Fabry model mice prevented Gb3 storage in the liver, kidneys, and heart and improved the pathological changes in these organs. Because this modified NAGA is hardly expected to cause an allergic reaction in Fabry disease patients, it is highly promising as a new and safe enzyme for ERT for Fabry disease.


Subject(s)
Enzyme Replacement Therapy/methods , Fabry Disease/drug therapy , alpha-N-Acetylgalactosaminidase/chemistry , alpha-N-Acetylgalactosaminidase/therapeutic use , Amino Acid Substitution , Animals , Binding Sites , CHO Cells , Catalysis , Cells, Cultured , Cricetinae , Cricetulus , Culture Media, Conditioned/chemistry , DNA, Complementary/metabolism , Disease Models, Animal , Drug Stability , Fabry Disease/enzymology , Fabry Disease/metabolism , Fibroblasts/drug effects , Fluorescent Dyes/metabolism , Galactosides/metabolism , Genetic Vectors , Humans , Hydrogen-Ion Concentration , Hymecromone/analogs & derivatives , Hymecromone/metabolism , Immunohistochemistry , Kidney/drug effects , Kidney/pathology , Kidney/ultrastructure , Liver/drug effects , Liver/pathology , Liver/ultrastructure , Mice , Mice, Knockout , Models, Molecular , Molecular Weight , Myocardium/pathology , Myocardium/ultrastructure , Recombinant Proteins/chemistry , Recombinant Proteins/isolation & purification , Recombinant Proteins/therapeutic use , Retroviridae/genetics , Transfection , Trihexosylceramides/metabolism , alpha-N-Acetylgalactosaminidase/genetics , alpha-N-Acetylgalactosaminidase/isolation & purification
SELECTION OF CITATIONS
SEARCH DETAIL
...