Navegando por Palavras-chave "Terapia de genes"
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- ItemEmbargoAvaliação do vetor retroviral bicistrônico Codificador de endostatina e interleucina-2 Para utilização em terapia gênica anti-tumoral(Universidade Federal de São Paulo (UNIFESP), 2010-07-28) Rocha, Flavia Gomes de Goes [UNIFESP]; Bellini, Maria Helena [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)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.
- ItemAcesso aberto (Open Access)Fator de crescimento do endotélio vascular na viabilidade do retalho musculofasciocutâneo transverso do reto do abdome, em ratos submetidos à nicotina(Universidade Federal de São Paulo (UNIFESP), 2009-07-30) Silveira, Tiago Santos [UNIFESP]; Ferreira, Lydia Masako [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)Introduction: Several factors can reduce the viability of the TRAM flap, among them the nicotine has been made responsible by the loss partial or total of these flaps. Objective: To evaluate the action of the Vascular endothelial Growth Factor in the viability of the Transverse Rectus Abdominis Musculocutaneous, in rats submitted to the nicotine. Methods: Sixty Wistar EPM-1 rats were used, adult males, weighing from 230 to 300g, randomized in 4 groups of 15 animals each: Group Control composed by animals that were submitted to the TRAM flap; Group Nicotine composed by animals that were nicotine exposed and submitted of TRAM flap; Group VEGF composed by animals submitted to the administration of VEGF plasmidial before the TRAM flap; and Group Nicotina+VEGF composed by animals that were exposed to the nicotine, trated with administration of VEGF and submitted to the TRAM flap. For analysis of the results they were done necrosis area and vascular density. Results: There was estatistic significant differentiates in the comparison among all of the groups, regarding the variables necrosis area and vascular density (p <0,05). Conclusion: The Vascular Endothelial Growth Factor increased the viability of the Transverse Rectus Abdominis Musculocutaneous, in rats submitted to the nicotine.
- ItemAcesso aberto (Open Access)Gene transfer to primary corneal epithelial cells with an integrating lentiviral vector(Conselho Brasileiro de Oftalmologia, 2010-10-01) Oliveira, Lauro Augusto De [UNIFESP]; Kim, Charles; Sousa, Luciene Barbosa De [UNIFESP]; Schwab, Ivan R.; Rosenblatt, Mark I.; Universidade Federal de São Paulo (UNIFESP); University of California Department of Ophthalmology and Visual Science; Weill Cornell Medical College Department of OphthalmologyPURPOSE: To evaluate the transfer of heterologous genes carrying a Green Fluorescent Protein (GFP) reporter cassette to primary corneal epithelial cells ex vivo. METHODS: Freshly enucleated rabbit corneoscleral tissue was used to obtain corneal epithelial cell suspension via enzymatic digestion. Cells were plated at a density of 5×10³ cells/cm² and allowed to grow for 5 days (to 70-80% confluency) prior to transduction. Gene transfer was monitored using fluorescence microscopy and fluorescence activated cell sorter (FACS). We evaluated the transduction efficiency (TE) over time and the dose-response effect of different lentiviral particles. One set of cells were dual sorted by fluorescence activated cell sorter for green fluorescent protein expression as well as Hoechst dye exclusion to evaluate the transduction of potentially corneal epithelial stem cells (side-population phenotypic cells). RESULTS: Green fluorescent protein expressing lentiviral vectors were able to effectively transduce rabbit primary epithelial cells cultured ex vivo. Live cell imaging post-transduction demonstrated GFP-positive cells with normal epithelial cell morphology and growth. The transduction efficiency over time was higher at the 5th post-transduction day (14.1%) and tended to stabilize after the 8th day. The number of transduced cells was dose-dependent, and at the highest lentivirus concentrations approached 7%. When double sorted by fluorescence activated cell sorter to isolate both green fluorescent protein positive and side population cells, transduced side population cells were identified. CONCLUSIONS: Lentiviral vectors can effectively transfer heterologous genes to primary corneal epithelial cells expanded ex vivo. Genes were stably expressed over time, transferred in a dose-dependence fashion, and could be transferred to mature corneal cells as well as presumable putative stem cells.
- ItemAcesso aberto (Open Access)Terapia gênica com VEGF para angiogênese na angina refratária: ensaio clínico fase I/II(Sociedade Brasileira de Cirurgia Cardiovascular, 2010-09-01) Kalil, Renato A. K.; Salles, Felipe Borsu De; Giusti, Imarilde Inês; Rodrigues, Clarissa Garcia; Han, Sang Won [UNIFESP]; Sant'anna, Roberto Tofani; Ludwig, Eduardo; Grossman, Gabriel; Prates, Paulo Roberto Lunardi; Sant'anna, João Ricardo Michelin; Teixeira Filho, Guaracy Fernandes; Nardi, Nance Beyer; Nesralla, Ivo Abrahão; Instituto de Cardiologia do Rio Grande do Sul/Fundação Universitária de Cardiologia; UFCSPA; Universidade Federal de São Paulo (UNIFESP); Instituto de Cardiologia do RS/Fundação Universitária de Cardiologia Laboratório de Cardiologia Molecular e CelularOBJECTIVE: Safety, feasibility and early myocardial angiogenic effects evaluation of transthoracic intramyocardial phVEGF165 administration for refractory angina in no option patients. METHODS: Cohort study, in which 13 patients with refractory angina under optimized clinical treatment where included, after cineangiograms had been evaluated and found unfeasible by surgeon and interventional cardiologist. Intramyocardial injections of 5mL solution containing plasmidial VEGF165 where done over the ischemic area of myocardium identified by previous SPECT/Sestamibi scan. Evaluations included a SPECT scan, stress test, Minnesotta QOL questionnaire and NYHA functional class and CCS angina class determinations. RESULTS: There were no deaths or new interventions during the study period. There were no significant variations in SPECT scans, QOL scores and stress tests results during medical treatment in the included patients. After the 3rd post operative month, there was improvement in SPECT segmental scores, SSS (18.38±7.51 vs. 15.31±7.29, P=0.003) and SRS (11.92±7.49 vs. 8.53±6.68, P=0.002). The ischemic area extension, however, had non-significant variation (23.38±13.12% vs. 20.08±13.88%, P=0.1). Stress tests METs varied from 7.66±4.47 pre to 10.29±4.36 METs post-op (P=0.08). QOL score improved from 48.23±18.35 pre to 30.15±20.13 post-op points (P=0.02). NYHA class was 3.15±0.38 pre vs. 1.77±0.83 post-op (P=0.001) and angina CCS class, 3.08±0.64 vs. 1.77±0.83 (P=0.001). CONCLUSIONS: Intramyocardial VEGF165 therapy for refractory angina, in this small trial of no option patients, resulted feasible and safe. Early clinical and scintilographic data showed improvements in symptoms and myocardial perfusion, with regression of ischemia severity in treated areas.
- ItemAcesso aberto (Open Access)Terapia gênica em distrofias hereditárias de retina(Conselho Brasileiro de Oftalmologia, 2009-08-01) Côco, Monique [UNIFESP]; Han, Sang Won [UNIFESP]; Sallum, Juliana Maria Ferraz; Centro Interdisciplinar de Terapia Gênica; Universidade Federal de São Paulo (UNIFESP)The inherited retinal dystrophies comprise a large number of disorders characterized by a slow and progressive retinal degeneration. They are the result of mutations in genes that express in either the photoreceptor cells or the retinal pigment epithelium. The mode of inheritance can be autosomal dominant, autosomal recessive, X linked recessive, digenic or mitochondrial DNA inherited. At the moment, there is no treatment for these conditions and the patients can expect a progressive loss of vision. Accurate genetic counseling and support for rehabilitation are indicated. Research into the molecular and genetic basis of disease is continually expanding and improving the prospects for rational treatments. In this way, gene therapy, defined as the introduction of exogenous genetic material into human cells for therapeutic purposes, may ultimately offer the greatest treatment for the inherited retinal dystrophies. The eye is an attractive target for gene therapy because of its accessibility, immune privilege and translucent media. A number of retinal diseases affecting the eye have known gene defects. Besides, there is a well characterized animal model for many of these conditions. Proposals for clinical trials of gene therapy for inherited retinal degenerations owing to defects in the gene RPE65, have recently received ethical approval and the obtained preliminary results brought large prospects in the improvement on patient's quality of life.
- ItemAcesso aberto (Open Access)Terapia gênica para osteoporose(Sociedade Brasileira de Ortopedia e Traumatologia, 2011-01-01) Costa, Rafael Pacheco da [UNIFESP]; Han, Sang Won [UNIFESP]; Pochini, Alberto de Castro [UNIFESP]; Reginato, Rejane Daniele [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)Osteoporosis is considered one of the most common and serious problems affecting the elderly population worldwide. It is a chronic and progressive disease, characterized by decreased bone mass and degeneration of the microarchitecture of the bone tissue. Gene therapy represents a new approach in osteoporosis treatment, and its main function is to restore the compromised function in the metabolism. This review aims to elucidate the main studies on gene therapy in recent years, in the medical databases, that use gene therapy for the treatment of osteoporosis in animal models, as well as the future prospects of this therapy. The majority of the studies use the BMP, PTH and OPG genes, in an attempt to reestablish bone mass. Despite the lack of new molecules, all genes employed in these studies have proven to be efficient in the treatment of the disease. The benefits that gene therapy will provide for patients in the future should contribute substantially to increasing the quality of life for the elderly. Soon, clinical trials involving humans will benefit individuals with osteoporosis.