RESUMO
BACKGROUND: Hepatocellular carcinoma (HCC) is a malignancy with poor survival outcome. New treatment options for the disease are needed. In this study, we identified and evaluated tumor vascular PLVAP as a therapeutic target for treatment of HCC. METHODS: Genes showing extreme differential expression between paired human HCC and adjacent non-tumorous liver tissue were investigated. PLVAP was identified as one of such genes with potential to serve as a therapeutic target for treatment of HCC. A recombinant monoclonal anti-PLVAP Fab fragment co-expressing extracellular domain of human tissue factor (TF) was developed. The potential therapeutic effect and toxicity to treat HCC were studied using a Hep3B HCC xenograft model in SCID mice. RESULTS: PLVAP was identified as a gene specifically expressed in vascular endothelial cells of HCC but not in non-tumorous liver tissues. This finding was confirmed by RT-PCR analysis of micro-dissected cells and immunohistochemical staining of tissue sections. Infusion of recombinant monoclonal anti-PLVAP Fab-TF into the main tumor feeding artery induced tumor vascular thrombosis and extensive tumor necrosis at doses between 2.5 µg and 12 µg. Tumor growth was suppressed for 40 days after a single treatment. Systemic administration did not induce tumor necrosis. Little systemic toxicity was noted for this therapeutic agent. CONCLUSIONS: The results of this study suggest that anti-PLVAP Fab-TF may be used to treat HCC cases for which transcatheter arterial chemoembolization (TACE) is currently used and potentially avoid the drawback of high viscosity of chemoembolic emulsion for TACE to improve therapeutic outcome. Anti-PLVAP Fab-TF may become a viable therapeutic agent in patients with advanced disease and compromised liver function.
Assuntos
Anticorpos Monoclonais/uso terapêutico , Carcinoma Hepatocelular/química , Carcinoma Hepatocelular/tratamento farmacológico , Proteínas de Transporte/análise , Células Endoteliais/química , Neoplasias Hepáticas/química , Neoplasias Hepáticas/tratamento farmacológico , Proteínas de Membrana/análise , Animais , Anticorpos Monoclonais/efeitos adversos , Antígenos de Superfície/imunologia , Carcinoma Hepatocelular/genética , Proteínas de Transporte/genética , Proteínas de Transporte/imunologia , Células Endoteliais/metabolismo , Feminino , Xenoenxertos , Humanos , Fígado/química , Neoplasias Hepáticas/genética , Masculino , Proteínas de Membrana/genética , Proteínas de Membrana/imunologia , Camundongos , Camundongos SCID , Terapia de Alvo Molecular , RNA Mensageiro/metabolismo , Proteínas Recombinantes/imunologiaRESUMO
BACKGROUND: Amiodarone is a class D drug given to treat arrhythmia, including pregnant women, but its effects on the developing heart have not been studied. Although some studies have suggested that this drug is safe for fetuses, they have been conducted on mothers with fetuses at or beyond six months of gestational age. RESULTS: The occurrence of valve defect was positively proportional to Amiodarone concentrations over 9 µM, but not lower than 6 µM. Ectopic overexpression of versican was observed at the atrioventricular canal of the Amiodarone-treated embryos at 15 µM (EC(50)). VE-cadherin (cdh5), normally downregulated at the endocardial cushion, was also ectopically overexpressed in the Amiodarone-treated embryos. Knockdown of either versican or cdh5 in the Amiodarone-treated embryos could rescue the valve defect caused by Amiodarone. CONCLUSIONS: By inducing versican ectopical overexpression, leading, in turn, to cdh5 ectopical overexpression, Amiodarone treatment causes failure of cardiac valve formation in zebrafish embryos.
Assuntos
Amiodarona/toxicidade , Antiarrítmicos/toxicidade , Valvas Cardíacas/efeitos dos fármacos , Teratogênicos/toxicidade , Animais , Antígenos CD/genética , Caderinas/genética , Embrião não Mamífero/efeitos dos fármacos , Embrião não Mamífero/embriologia , Valvas Cardíacas/embriologia , Versicanas/genética , Peixe-Zebra/embriologia , Proteínas de Peixe-Zebra/genéticaRESUMO
Shannon information in the genomes of all completely sequenced prokaryotes and eukaryotes are measured in word lengths of two to ten letters. It is found that in a scale-dependent way, the Shannon information in complete genomes are much greater than that in matching random sequences--thousands of times greater in the case of short words. Furthermore, with the exception of the 14 chromosomes of Plasmodium falciparum, the Shannon information in all available complete genomes belong to a universality class given by an extremely simple formula. The data are consistent with a model for genome growth composed of two main ingredients: random segmental duplications that increase the Shannon information in a scale-independent way, and random point mutations that preferentially reduces the larger-scale Shannon information. The inference drawn from the present study is that the large-scale and coarse-grained growth of genomes was selectively neutral and this suggests an independent corroboration of Kimura's neutral theory of evolution.
Assuntos
Mapeamento Cromossômico/métodos , Análise Mutacional de DNA/métodos , DNA/genética , Evolução Molecular , Armazenamento e Recuperação da Informação/métodos , Modelos Genéticos , Alinhamento de Sequência/métodos , Análise de Sequência de DNA/métodos , Algoritmos , Biologia Computacional/métodos , DNA/química , Variação Genética/genética , Modelos EstatísticosRESUMO
Shannon information in the genomes of all completely sequenced prokaryotes and eukaryotes are measured in word lengths of two to ten letters. It is found that in a scale-dependent way, the Shannon information in complete genomes are much greater than that in matching random sequences - thousands of times greater in the case of short words. Furthermore, with the exception of the 14 chromosomes of Plasmodium falciparum, the Shannon information in all available complete genomes belong to a universality class given by an extremely simple formula. The data are consistent with a model for genome growth composed of two main ingredients: random segmental duplications that increase the Shannon information in a scale-independent way, and random point mutations that preferentially reduces the larger-scale Shannon information. The inference drawn from the present study is that the large-scale and coarse-grained growth of genomes was selectively neutral and this suggests an independent corroboration of Kimura's neutral theory of evolution.