Avaliação do vetor retroviral bicistrônico Codificador de endostatina e interleucina-2 Para utilização em terapia gênica anti-tumoral. / Evaluation of retroviral vector encoding bicistônico of endostatin and interleukin-2 for use in anti-tumor gene therapy

AUTOR(ES)
FONTE

IBICT - Instituto Brasileiro de Informação em Ciência e Tecnologia

DATA DE PUBLICAÇÃO

28/07/2010

RESUMO

Gene therapy has been used in preclinical studies and clinical trials in order to assuage or cure a disease. Retroviral vectors are tools for gene transfer are widely used. Bicistronic vectors are attractive alternatives for treatment of complex disease. Endostatin (ES), the C-terminal fragment of collagen XVIII, is a potent angiogenesis inhibitor. At present, ES has been widely used in a variety of experimental tumor models and clinical trial. Immunotherapy, with interleukin-2 (IL-2), has been used as adjuvant treatment for tumors in several preclinical studies and clinical trials. The objective of this project was evaluated an IRES-based bicistronic retroviral vector encoding ES and IL-2, in anti-tumor gene therapy. In this study, the potential of Bicistronic therapy on activation of tumorinfiltrating lymphocytes in metastatic renal cell carcinoma (mRCC) was examined, using an orthotopic metastatic mouse model. Renca cells were injected into the renal subcapsule of BALB/C mice. After seven days, the nefrectomy was performed. Then, the animals were randomly divided into two groups (10 mice/group): control and the second group of mice received a subcutaneous inoculation of NIH/3T3-LendSN-IRES-IL-2 cells. Ten days after the nefrectomy, the animals were exsanguinated and killed. In the survival studies, daily mice were monitored daily, until they died. At the end of the in vivo experiment, serum levels of IL-2 and ES were measured, the lung was weighed, and number of nodule mestastatic, nodule area, microvascular area (MVA), proliferation of RenCa cells infiltrating tumor cells, plus tumors-infiltrating lymphocytes were evaluated. There was a significant decrease in the number of metastatic lung nodules, wet weight, lung nodule area, MVA and proliferation of Renca cells in the Bicistronic-treated group compared to the control group. These significant differences revealed an antitumor effect on the bicistronic (ES+IL-2) treatment. Subcutaneous inoculation of NIH/3T3- LendSNIRES-IL-2 cells resulted in an increase in ES and IL-2 levels and in the infiltration of CD4, CD4 producer IFNg, CD8, CD8 producer IFNg and NK (CD49b) cells in the treated group. The Bicistronic therapy Kaplan Meier survival curves showed that the probability of survival was significantly higher for mice treated (log-rank test, P=0.0016). Concluding, retroviral ES and IL-2 gene transfer led to secretion of functional ES and IL-2 that were sufficiently active to inhibit tumor angiogenesis and tumor growth, increasing the infiltration of immune cells.

ASSUNTO(S)

terapia de genes endostatinas interleucina-2 nefrologia

ACESSO AO ARTIGO

Documentos Relacionados