RESUMO
Cytochrome P450 (CYP450) is the leading enzyme in the biotransformation of most psychotropic drugs. CYP450 gene polymorphisms determine a patient's endophenotype with respect to the activity of enzymes of the family and affect the metabolism of prescribed antipsychotics and antidepressants. Categorizing patients by endophenotype during genotyping is likely to help simplify the selection of therapy in clinical practice. Co-prescribing drugs that may be inhibitors or inducers of CYP450 isoforms, in turn, may lead to adverse reactions or no effect of therapy. The article presents a compilation of known pharmacogenetic recommendations regarding the four major endophenotypes of metabolizers.
Assuntos
Antipsicóticos , Humanos , Antipsicóticos/efeitos adversos , Sistema Enzimático do Citocromo P-450/genética , Psicotrópicos , Antidepressivos/efeitos adversos , Farmacogenética/métodos , Isoformas de Proteínas/genética , GenótipoRESUMO
Glioblastoma is a primary brain tumor and one of the most aggressive malignant neoplasms. The prognosis remains poor with a short survival period after diagnosis even in the case of timely detection and early treatment with the use of advanced chemotherapy, radiation therapy and surgical treatment. In this regard, the research of the main pathogenetic links in the glioblastoma development continues. The current focus is on studying the molecular characteristics of tumours, including the analysis of extracellular vesicles, which play an essential role in intercellular communication processes. In this review, in order to provide up-to-date information on the role of extracellular vesicles in the diagnosis and therapy of gliomas, the analysis of the achieved results of Russian and foreign research related to this area has been carried out. The main goal of this review is to describe the features of extracellular vesicles as the containers and glioma marker transporters, as well as nucleic acids used in diagnosis and therapy.
Assuntos
Neoplasias Encefálicas , Vesículas Extracelulares , Glioblastoma , Glioma , Humanos , Glioblastoma/diagnóstico , Neoplasias Encefálicas/diagnóstico , Neoplasias Encefálicas/terapia , Biomarcadores Tumorais/análise , Glioma/diagnóstico , Glioma/terapia , Glioma/patologia , Vesículas Extracelulares/química , Vesículas Extracelulares/patologiaRESUMO
OBJECTIVE: To investigate the effects of diet on the gut microbiota and to assess the relationship of these factors with depression. MATERIAL AND METHODS: Microorganisms that predominate in depressed patients were identified and associations of the identified organisms with the patients' diet were performed. Fourteen depressed patients and 14 healthy volunteers with the same socio-demographic parameters were included in the study. The Hamilton Depression Scale, Generalized Anxiety Disorder Questionnaire, and the Center for Epidemiologic Studies Questionnaire were used. RESULTS: Erysipelatoclostridium and Clostridium innocuum species were 11.3 and 14.4 times higher in depressed patients compared with healthy controls. Fusicatenibacter saccharivorans, Faecalibacterium prausnitzii and Roseburia faecis species, as well as members of the genus Roseburia were statistically significantly more abundant in the healthy volunteers group (6.5, 2.14, 8.75 and 5.2 times more frequently compared to patients). The presence of these microorganisms was correlated with dietary components. CONCLUSION: Our study revealed groups of microorganisms that differ in healthy volunteers and depressed patients. The association of these microorganisms with the diet was shown, which partially confirmed the influence of a «healthy diet¼ on the development of depressive disorders.
Assuntos
Microbioma Gastrointestinal , Depressão , Dieta , Fezes/microbiologia , Humanos , RNA Ribossômico 16SRESUMO
Dendritic cell-based vaccines are an intensively studied active immunotherapy technology. Aim of this article is to review the results of the key clinical studies of such vaccines in the treatment of neuro-oncological diseases. Their effectiveness was studied most widely in the treatment of malignant glial tumors, the study went from experimental work to phase III clinical studies, preliminary results of which indicate some positive results of this immunotherapy method in adults. Currently, emphasis is also being placed on the identification of clinical and immunological correlates of the patient's response to therapy and on the search for new antigens for sensitization of dendritic cells Studies of dendritic cell vaccines also include a number of other neuro-oncological diseases. A separate part of this article is devoted to the treatment of intracerebral tumors in children, for example, medulloblastomas and gliomas of the pons. In addition, the potential use of dendritic cell vaccines for intracerebral metastases is considered.
Assuntos
Neoplasias Encefálicas/terapia , Vacinas Anticâncer/uso terapêutico , Neoplasias Cerebelares , Glioma , Adulto , Criança , Células Dendríticas/imunologia , HumanosRESUMO
AIM: To investigate serotonin and catecholamine levels in people with paraphilic disorders and identify correlations between transmitter dysfunction and clinical signs of paraphilic disorders. MATERIAL AND METHODS: Fifteen men with paraphilic disorders were studied using clinical-psychopathological, sexological, biochemical and statistical methods. RESULTS: There were an increase in the levels of serotonin and norepinephrine and a decrease in the concentration of DOPAC (3,4-dihydroxyphenylacetic acid) in the urine of patients with paraphilic disorders. The concentrations of serotonin and norepinephrine are correlated with obsessive disturbances. The level of DOPAC was associated with affective and dissociative disorders. CONCLUSION: The relationships between biochemical and psychopathological signs suggest a role of biological mechanisms in the organization of abnormal sexual behavior. Correlations between psychopathological phenomena and DOPAC indicate a key role of central dopamine in the pathogenesis of paraphilic disorders and disturbances of conscious regulation of behavior.
Assuntos
Norepinefrina , Transtornos Parafílicos , Serotonina , Dopamina/metabolismo , Humanos , Masculino , Neurotransmissores , Norepinefrina/metabolismo , Transtornos Parafílicos/metabolismo , Transtornos Parafílicos/fisiopatologia , Serotonina/metabolismoRESUMO
We obtained the morphologically, cytofluorometrically, and functionally mature dendritic cells from rats that were pulsed with antigens of the C6 glioma tissue extract. The concentrations of angiogenesis antigens (VEGF, VEGFR-1, and VEGFR-2) and periglioma zone proteins (GFAP, connexin 43, and BSAT1) in the pulsing extract were measured by ELISA. Our results drove us to a conclusion that despite mature phenotype of pulsed dendritic cell, the antigenic composition of glioma tissue extracts should be modified.
Assuntos
Antígenos de Neoplasias/genética , Neoplasias Encefálicas/química , Misturas Complexas/farmacologia , Células Dendríticas/efeitos dos fármacos , Glioma/química , Animais , Antígenos CD/genética , Antígenos CD/imunologia , Antígenos de Neoplasias/imunologia , Química Encefálica , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patologia , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Misturas Complexas/química , Conexina 43/genética , Conexina 43/imunologia , Células Dendríticas/citologia , Células Dendríticas/imunologia , Regulação da Expressão Gênica/imunologia , Proteína Glial Fibrilar Ácida/genética , Proteína Glial Fibrilar Ácida/imunologia , Glioma/genética , Glioma/metabolismo , Glioma/patologia , Humanos , Imunofenotipagem , Transplante de Neoplasias , Proteínas de Transporte de Cátions Orgânicos/genética , Proteínas de Transporte de Cátions Orgânicos/imunologia , Cultura Primária de Células , Ratos , Técnicas Estereotáxicas , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/imunologia , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/genética , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/imunologia , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/genética , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/imunologiaRESUMO
The efficiency of conventional chemotherapy for aggressive tumors in the CNS remains low and new strategies for the targeted delivery of anti-tumor substances are now actively developed. Pegylated liposomes covalently conjugated with monoclonal antibodies to VEGF synthesized by us are nanoparticle characterized by narrow size distribution and high dispersion stability. Immunochemical activity of antibodies after conjugation was 70% of initial level. The anti-VEGF liposomes developed by us were highly specific for VEGF(+) tumor cells (in vitro and in vivo). Intravenous injection of VEGF-liposomes to rats with intracranial C6 glioma was followed by their specific accumulation in the malignant tissues and engulfment by glioma cells, which attested to target delivery and selective accumulation of anti-VEGF-liposomes in the brain tumor. Thus, the use of targeting molecules can significantly increase the distribution and efficiency of delivery of nanocontainers to a tumor characterized by hyperexpression of the target proteins.
Assuntos
Anticorpos Monoclonais/uso terapêutico , Neoplasias Encefálicas/tratamento farmacológico , Glioma/tratamento farmacológico , Lipossomos/administração & dosagem , Lipossomos/química , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores , Animais , Anticorpos Monoclonais/administração & dosagem , Linhagem Celular Tumoral , Feminino , Ratos , Ratos Wistar , Fator A de Crescimento do Endotélio Vascular/imunologiaRESUMO
Antitumor efficiencies of monoclonal antibodies to connexin-43 second extracellular loop (MAbE2Cx43), temozolomide, and fractionated γ-irradiation in the monotherapy mode and in several optimized combinations were studied in Wistar rats with induced C6 glioma. The survival of animals with glioma and the dynamics of intracerebral tumor development were evaluated by MRT. Temozolomide monotherapy (200 mg/m(2)) and isolated radiotherapy in a total dose of 36 Gy shifted the survival median from 28 days (no therapy) to 34 and 38 days, respectively; 100% animals died under conditions of temozolomide monotherapy and radiotherapy. Monotherapy with MAbE2Cx43 in a dose of 5 mg/kg led to significant regression of the tumor (according to MRT data), cure of 19.23% animals with glioma, and prolongation of the survival median to 39.5 days after tumor implantation. Combined therapy with MAbE2Cx43 and temozolomide completely abolished the antitumor effect (survival median 29 days). Treatment with MAbE2Cx43 in combination with radiotherapy was associated with mutual boosting of the therapeutic efficiencies, leading to a significant inhibition of tumor development and prolongation of the survival median to 60 days. The mechanism of tumorsuppressive activity of the antibodies could be due to connexon blockade in Cx43-positive glioma cells in the peritumor invasion zone. Higher efficiency of combined therapy was presumably due to the increase in blood-brain barrier permeability for antibodies after irradiation of the brain and to additional inhibitory effect of antibodies towards radioresistant migrating glioma cells. The results suggested that MAbE2Cx43 could be effective as the first-line drug in combined therapy for poorly differentiated gliomas.
Assuntos
Anticorpos Monoclonais/farmacologia , Neoplasias Encefálicas/terapia , Conexina 43/imunologia , Dacarbazina/análogos & derivados , Raios gama/uso terapêutico , Glioblastoma/terapia , Animais , Antineoplásicos Alquilantes/metabolismo , Antineoplásicos Alquilantes/farmacologia , Barreira Hematoencefálica/efeitos da radiação , Neoplasias Encefálicas/imunologia , Neoplasias Encefálicas/mortalidade , Neoplasias Encefálicas/patologia , Permeabilidade Capilar/efeitos da radiação , Terapia Combinada/métodos , Conexina 43/química , Dacarbazina/metabolismo , Dacarbazina/farmacologia , Esquema de Medicação , Feminino , Glioblastoma/imunologia , Glioblastoma/mortalidade , Glioblastoma/patologia , Camundongos , Camundongos Endogâmicos BALB C , Transplante de Neoplasias , Estrutura Terciária de Proteína , Ratos , Ratos Wistar , Técnicas Estereotáxicas , Análise de Sobrevida , Temozolomida , Carga Tumoral/efeitos dos fármacos , Carga Tumoral/efeitos da radiaçãoRESUMO
Fluorescent diagnosis was first proposed in the early XX century and has been used in neurosurgery for about 15 years. The method relies on selective accumulation of strongly fluorescent protoporphyrin IX in tumor cells. Over the past years, the method of intraoperative fluorescence diagnosis has occupied its niche in many neurosurgical clinics around the world and is now used for fast intraoperative diagnosis in brain tumor surgery. However, the efficiency of fluorescent intraoperative diagnosis using 5-aminolevulinic acid is 80-90% and 58.8% for surgery of Grade III-IV and I-II gliomas, respectively. One of the methods to improve the efficiency of fluorescent diagnosis is to use vector systems for delivering fluorescent drugs into the tumor. This paper reports the results of an experimental study of systems for delivering fluorescent agents (protoporphyrin IX, Alexa 488, Alexa 660) using connexin-43 antibodies in rats with transplanted C6 glioma.
Assuntos
Anticorpos Monoclonais , Neoplasias Encefálicas/cirurgia , Conexina 43/imunologia , Fluorescência , Glioma/cirurgia , Cuidados Intraoperatórios/métodos , Procedimentos Neurocirúrgicos/métodos , Animais , Neoplasias Encefálicas/imunologia , Neoplasias Encefálicas/patologia , Linhagem Celular Tumoral , Corantes Fluorescentes , Glioma/imunologia , Glioma/patologia , Gradação de Tumores , Transplante de Neoplasias , Neuronavegação , Fármacos Fotossensibilizantes , Ratos , Espectrometria de FluorescênciaRESUMO
cDNA extracellular Ig-like domains I-III of vascular endothelial growth factor receptor 2 (VEGFR2) was cloned in an expressing vector pET_32a. Western blotting showed immunochemical identity of recombinant VEGFR2I-III produced by prokaryotic expression system to the native receptor. BALB/c mice were immunized with VEGFR2I-III for obtaining specific antibodies to VEGFR2. Hybridomas producing monoclonal antibodies were selected by ELISA, Western blotting, and immunocytochemical assay. Thus, we obtained hybridoma producing monoclonal antibodies to VEGFR2 that selectively interact with both recombinant and native extracellular fragment of the receptor.
Assuntos
Anticorpos Monoclonais/imunologia , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/imunologia , Animais , Feminino , Células Endoteliais da Veia Umbilical Humana , Humanos , Camundongos , Camundongos Endogâmicos BALB CRESUMO
The review is devoted to a relatively young direction in therapy of malignant gliomas, which is based on applying monoclonal antibodies against tumour-associated antigens. The current data on efficacy of main therapeutic agents in clinical practice or clinical trials concerning high-grade gliomas, especially glioblastoma multiforme, is summarized. Of particular interest is bevacizumab, a humanized monoclonal antibody against vascular endothelial growth factor (anti-VEGF), which is widely used in glioblastoma. Major clinical trials devoted to bevacizumab monotherapy and combinations of bevacizumab with other therapeutic modalities in primary and recurrent glioblastoma conducted since 2006 till now are reviewed. The results ofexperimental and clinical application ofmonoclonal antibodies against epidermal growth factor receptor (EGFR) and its mutant variant EGFRvIII are analyzed, showing the most significant clinical effectiveness of nimotuzumab--a humanized monoclonal antibody. Significant part of the review is devoted to discussion of experimental and clinical data concerning efficacy of antibodies against VEGF receptor 2, platelet-derived growth factor receptor α, hepatocyte growth factor and its receptor c-Met. Unbiassed analysis of clinical trials on monoclonal antibodies does not allow us to conclude that passive immunotherapy directed against antigens listed above can significantly improve the overall survival of patients suffering from glioblastoma multiforme. This finding has encouraged us to mention several alternative approaches to passive immunotherapy and to list several prospective antigens for developing monoclonal antibodies.
Assuntos
Anticorpos Monoclonais/uso terapêutico , Neoplasias do Sistema Nervoso Central/tratamento farmacológico , Glioma/tratamento farmacológico , Animais , Anticorpos Monoclonais/farmacologia , Anticorpos Monoclonais Humanizados/farmacologia , Anticorpos Monoclonais Humanizados/uso terapêutico , Bevacizumab , Neoplasias Encefálicas/tratamento farmacológico , Neoplasias Encefálicas/patologia , Neoplasias do Sistema Nervoso Central/patologia , Receptores ErbB/antagonistas & inibidores , Receptores ErbB/genética , Glioma/patologia , Humanos , Camundongos , Mutação , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/farmacologia , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores , Fator A de Crescimento do Endotélio Vascular/imunologia , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/antagonistas & inibidores , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/antagonistas & inibidoresRESUMO
We studied the effects of single intravenous injection of antibodies to brain-specific transmembrane anion transporter (BSAT1; 5 mg/kg) to pregnant rats (gestation day 10) on cognitive functions and behavior of their progeny. One of major functions of BSAT1 (or Oatp1c1) is specific transport of thyroxin across the blood-brain barrier. Female rats of two control groups were injected with non-specific Ig and 0.9% NaCl. The progeny of rats receiving monoclonal antibodies to BSAT1 demonstrated memory impairment in the Y-maze, novel object recognition test, passive avoidance test, and Morris water maze test in comparison with the control group. Our findings suggest that single injection of monoclonal antibodies to BSAT1 during the prenatal period was followed by cognitive impairments, which were probably related to thyroxin deficiency in the nervous tissue.
Assuntos
Proteínas de Transporte de Cátions Orgânicos/metabolismo , Efeitos Tardios da Exposição Pré-Natal/psicologia , Animais , Anticorpos Monoclonais/farmacologia , Aprendizagem da Esquiva , Cognição , Feminino , Aprendizagem em Labirinto , Memória de Curto Prazo , Proteínas de Transporte de Cátions Orgânicos/antagonistas & inibidores , Gravidez , Efeitos Tardios da Exposição Pré-Natal/metabolismo , RatosRESUMO
Organ, cellular, and subcellular localization of brain-specific anion transporter BSAT1 was studied in rats using antibodies to the extracellular fragment (451-557 a.a). The antibodies were shown to recognize the antigen predominantly localized in the nervous tissue, tumors of glial origin, and primordial ovarian follicles. The absence of BSAT1 immunofluorescence signal in kidney and liver sections and accumulation of (125)I labeled antibodies to BSAT1 in these organs indicate that these antibodies do not cross-react with the most common isoforms of OATP expressed in these organs. Analysis of the cellular localization suggests that in the brain, BSAT1 is localized predominantly in astrocytes, but not in endothelial cells, as was previously reported. Laser scanning confocal microscopy with a set of relevant trackers revealed membrane localization of BSAT1. Taking into account the data on the of localization, we can conclude that antibodies to BSAT1 451-557 can be used for basic research of the transport of thyroxin and prostaglandins across the blood brain barrier and for testing the systems for targeted transport of diagnostic preparations and drugs across the blood brain barrier, e.g. to astroglial tumors.
Assuntos
Encéfalo/metabolismo , Proteínas de Transporte de Cátions Orgânicos/metabolismo , Sequência de Aminoácidos , Animais , Animais não Endogâmicos , Anticorpos Monoclonais Murinos , Astrócitos/metabolismo , Encéfalo/citologia , Linhagem Celular Tumoral , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Células HEK293 , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Hibridomas , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Especificidade de Órgãos , Proteínas de Transporte de Cátions Orgânicos/química , Proteínas de Transporte de Cátions Orgânicos/imunologia , Fragmentos de Peptídeos/imunologia , Transporte Proteico , RatosRESUMO
Glioblastoma (GBM) is the most common type of primary brain cancer that characterized by poor prognosis due to the rapid progression, active angiogenesis, enhanced tumor cell invasion and the emergence of resistance toward conventional therapy. In this connection, nowadays, new approaches for selective inhibition of crucial steps in tumor progression are actively developing. The key feature of tumor growth and development is angiogenesis. VEGF and its receptor VEGFR2 play the pivotal role in regulation of tumor vessel formation. Therefore, VEGFR2, as the main receptor of VEGF's pro-angiogenic signal transducer, is a promising molecular target for anti-angiogenic therapy. There is evidence that inhibitors of VEGF and VEGFR2 reduce endothelial cell proliferation, migration and survival that lead to regression of vessel density and decrease vascular permeability, thereby slowing tumor growth. Currently, a number of VEGFR2 inhibitors are under clinical trials (ramucirumab, cediranib) and several were approved (sunitinib, sorafenib). Despite the promising results of preclinical studies, the efficacy of antiangiogenic drugs in the clinical practice is significantly lower, mainly, due to rapid adaptation of malignant cells that consists of alternative pro-angiogenic pathways activation, recruitment of endothelial progenitor cells from bone marrow and increasing of the invasive growth. Given the diversity of pro-angiogenic mechanisms, enhancement of the efficacy of tumor therapy could be achieved by specific inhibition of VEGFR2 functions that will be supplemented by other antiangiogenic drugs (anti-VEGF,-PIGF,-HIF1alpha). In addition, multitargeting therapy should focus on the combined inhibition of angiogenesis, invasion, metastasis, proliferation and survival of tumor cells.
Assuntos
Inibidores da Angiogênese/uso terapêutico , Neoplasias Encefálicas , Neovascularização Patológica/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Neoplasias Encefálicas/irrigação sanguínea , Neoplasias Encefálicas/tratamento farmacológico , Neoplasias Encefálicas/metabolismo , Proliferação de Células/efeitos dos fármacos , Humanos , Fator A de Crescimento do Endotélio Vascular/efeitos dos fármacosRESUMO
The results of fundamental and applied studies of blood-brain barrier had been conducted by authors during the last 10 years are summarized in the publication. The molecular anatomy of barrier microvessels, as well as promising markers of BBB and other proteins involved in barrier functions are discussed. Via in vitro experiments with endothelial cells of cerebral microvessels we characterized the basic conditions required for adequate BBB modeling. The in vivo data of BBB permeability for macromolecules in normal and different pathological process is including radiation injury, hyperosmotic shock, and nervous tissue ischemia are properly described. A particular attention was focused upon the experimental studies of the permeability and functional reorganization of barrier endothelium during tumor neoangiogenesis. We detected a dramatically increased permeability of neoplastic microvessels both for horseradish peroxidase/serum albumin and labeled monoclonal antibodies. The increased tumor permeability for IgG and the overexpression of target antigens in tumor tissue and peritumoral zone make possible the targeted delivery of diagnostics and therapeutic agents into the tumor by means of monoclonal antibodies.
Assuntos
Pesquisa Biomédica , Barreira Hematoencefálica/fisiologia , Endotélio Vascular/metabolismo , Neuroglia/metabolismo , Transporte Biológico , HumanosRESUMO
Female BALB/c mice were subcutaneously immunized with recombinant VEGF-164. After 3 immunization cycles, splenic B cells from immunized mouse were fused with immortalized myeloma culture SP2/0-Ag14 cells. Screening of hybrid cells producing anti-VEGF antibodies was performed by ELISA and immunocytochemical analysis on cultured C6 glioma cells. Subsequent cloning yielded hybridoma stably expressing monoclonal anti-VEGF antibodies recognizing recombinant and native VEGF.
Assuntos
Anticorpos Monoclonais/biossíntese , Fator A de Crescimento do Endotélio Vascular/imunologia , Animais , Anticorpos Monoclonais/imunologia , Linhagem Celular Tumoral , Ensaio de Imunoadsorção Enzimática , Feminino , Imuno-Histoquímica , Camundongos , Camundongos Endogâmicos BALB CRESUMO
We studied the effect of intravenous administration of monoclonal antibodies to the second extracellular loop of connexin 43 (MAbE2Cx43) on the dynamics of glioma growth and survival of experimental animals. Morphometric analysis of magnetic resonance imaging data showed that weekly intravenous administration of MAbE2Cx43 in a dose of 5 mg/kg significantly reduced glioma volume starting from day 21 after tumor implantation. By day 29, the mean volume of glioma in the experimental group (therapy with specific antibodies) was 2-fold lower than in controls. Deceleration of glioma growth in rats receiving MAbE2Cx43 was accompanied by a significant prolongation of rat lifespan (according to Kaplan-Meier test) and even led to complete recovery without delayed relapses in 19.23% animals. The mechanism of tumor-suppressing effects of antibodies can be related to inhibition of specific functions of connexin 43 in glioma cells in the peritumoral zone.
Assuntos
Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/uso terapêutico , Conexina 43/imunologia , Glioma/tratamento farmacológico , Animais , Feminino , Camundongos , Ratos , Ratos WistarRESUMO
Brain-specific anion transporter BSAT1 extracellular fragment (451-557) cDNA was cloned in a vector for prokaryotic expression, a producer E. coli strain was obtained, and recombinant extracellular fragment BSAT1(451-557)was purified and used for immunization of BALB/c mice. Splenic cells from mice with verified immune response were used for hybridoma generation. Several hybridoma clones producing monoclonal antibodies to BSAT1 extracellular fragment were selected. Antibody specificity was confirmed by ELISA, immunoblotting with recombinant BSAT1(451-557), and immunofluorescent BSAT1 assay on rat brain sections and cultured HEK293 cells. It was demonstrated that the obtained antibodies specifically bind native rat and human BSAT1 and can be used in both fundamental studies of structures forming the blood-brain barrier and development of targeted transport of diagnostic preparations and drugs across the blood-brain barrier.
Assuntos
Anticorpos Monoclonais/biossíntese , Encéfalo/metabolismo , Expressão Gênica , Transportadores de Ânions Orgânicos/genética , Sequência de Aminoácidos , Animais , Anticorpos Monoclonais/imunologia , Clonagem Molecular , Escherichia coli/genética , Vetores Genéticos , Humanos , Hibridomas/imunologia , Hibridomas/metabolismo , Imunização , Imunoensaio , Camundongos , Dados de Sequência Molecular , Transportadores de Ânions Orgânicos/química , Transportadores de Ânions Orgânicos/metabolismo , Proteínas de Transporte de Cátions Orgânicos , Ligação Proteica , Estrutura Terciária de Proteína , Ratos , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Sensibilidade e EspecificidadeRESUMO
Solid tumor progression largely depends on vascularization and angiogenesis in the malignant tissue. The most prominent among all proangiogenic factors is vascular endothelium growth factor (VEGF). VEGF suppression leads to retrogression of neoplastic vessels and tumor growth restriction. Clinical trials of complex antiangiogenic and chemical therapy of different neoplastic tumors have shown promising results. Nowadays bevacizumab is widely used in breast cancer, colorectal cancer and II-IV stage of malignancy gliomas treatment. Unfortunately, in the majority of cases antiangiogenic treatment led not to full recovery, but only to tumor development restriction. Resistance mechanisms include potentiating of alternative proangiogenic signaling pathways and activation of malignant cell invasive population.