RESUMEN
OBJECTIVE: Tofogliflozin is an oral hypoglycemic agent with a novel mechanism of action that reduces blood glucose levels by promoting glucose excretion in urine, achieved by selectively inhibiting sodium-glucose co-transporter 2 (SGLT2). We evaluated the effects of several selected anti-type 2 diabetes mellitus (T2DM) drugs-glimepiride, metformin, sitagliptin, pioglitazone, miglitol, nateglinide, and voglibose-on the pharmacokinetics and pharmacodynamics of tofogliflozin, and the effects of tofogliflozin on the pharmacokinetics of these anti-T2DM drugs in healthy male volunteers. METHODS: A single dose of either tofogliflozin alone, one of the anti-T2DM drugs alone, or co-administration of tofogliflozin and the anti-T2DM drug was administered to 108 healthy men. Cmax, AUCinf, and cumulative urine glucose excretion after co-administration of tofogliflozin and each of the anti-T2DM drugs was evaluated relative to the values of those parameters after administration of each drug alone. RESULTS: None of the anti-T2DM drugs had any effect on tofogliflozin exposure. Tofogliflozin had no or little effect on the exposure of any anti-T2DM drug. No anti-T2DM drug had any major effect on the cumulative urine glucose excretion induced by tofogliflozin. There were no safety concerns evident after administration of any drug alone or in co-administration. CONCLUSIONS: Neither the pharmacokinetics nor the pharmacodynamics of tofogliflozin was affected by any of the anti-T2DM drugs evaluated in this study, nor was the pharmacokinetics of any of the anti-T2DM drugs affected by tofogliflozin in healthy male volunteers.
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Compuestos de Bencidrilo/farmacología , Compuestos de Bencidrilo/farmacocinética , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Glucósidos/farmacología , Glucósidos/farmacocinética , Hipoglucemiantes/farmacología , Hipoglucemiantes/farmacocinética , Inhibidores del Cotransportador de Sodio-Glucosa 2 , 1-Desoxinojirimicina/análogos & derivados , 1-Desoxinojirimicina/farmacocinética , 1-Desoxinojirimicina/farmacología , Adulto , Ciclohexanos/farmacocinética , Ciclohexanos/farmacología , Diabetes Mellitus Tipo 2/sangre , Interacciones Farmacológicas , Glucosa/análisis , Voluntarios Sanos , Humanos , Inositol/análogos & derivados , Inositol/farmacocinética , Inositol/farmacología , Masculino , Metformina/farmacocinética , Metformina/farmacología , Persona de Mediana Edad , Nateglinida , Fenilalanina/análogos & derivados , Fenilalanina/farmacocinética , Fenilalanina/farmacología , Pioglitazona , Fosfato de Sitagliptina/farmacocinética , Fosfato de Sitagliptina/farmacología , Compuestos de Sulfonilurea/farmacocinética , Compuestos de Sulfonilurea/farmacología , Tiazolidinedionas/farmacocinética , Tiazolidinedionas/farmacología , Orina/química , Adulto JovenRESUMEN
The purpose of the study was to compare the peripheral retinal sensitivity of the visual field between professional soccer players and age-gender matched non-athlete subjects. All participants underwent a complete eye evaluation. The visual field was evaluated with the achromatic program 60-4 from the Humphrey automated perimetry. The binocular visual field was created with the best location model. It was divided into 4 quadrants (left superior, right superior, left inferior, and right inferior) and compared between groups. The study group comprised 29 professional male football players and the control group comprised 26 age-matched male non-athletes. Mean age was 25.8±4.7 years in the study group and 26.3±5.1 for controls. The average of retina sensitivity in the left inferior and right inferior quadrants was higher in the study group (27.2±1.2 dB and 27.0±1.4 dB) as compared to controls (26.1±1.9 dB and 25.5±2.1 dB). (Student's t test, P=0.011 and P=0.004, respectively). In this small cohort, professional soccer players presented higher retina sensitivity in the inferior quadrants when compared to non-athletes.
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Atletas , Retina/fisiología , Fútbol , Campos Visuales/fisiología , Adolescente , Adulto , Estudios Transversales , Humanos , Masculino , Pruebas del Campo Visual , Adulto JovenRESUMEN
Measurement of serum glycopeptidolipid core IgA antibody (GPL antibody) was recently reported to show a high sensitivity and specificity for diagnosing Mycobacterium avium-intracellulare complex (MAC) pulmonary disease (MAC-PD), but its clinical value has not been confirmed. This study aims to evaluate the seropositive rate in patients with suspected MAC-PD based on chest computed tomography (CT), and to examine whether GPL antibody reflects the extent of lung involvement on CT or the number of bacteria in sputum, retrospectively. Among 66 patients with suspected MAC-PD on CT, 36 patients were negative for MAC by culture and 30 were positive. Sputum grades of MAC were evaluated by fluorochrome microscopy of sputum smears. The lungs were divided into six regions to assess the extent of disease. Serum levels of GPL antibody were measured with an enzyme immunoassay (cut-off value >0.7 U/ml). The GPL antibody positive rate was 19.4% among patients who were negative for MAC by culture versus 73.3% among culturepositive patients. The serum level of GPL antibody was significantly correlated with the sputum smear grade (r=0.43, p less than 0.05) and was also correlated with the number of lung regions showing MAC-PD features on CT (r=0.43, less than 0.05). Some MAC-PD patients may have CT features of MAC with positive level of GPL antibody, although the diagnosis cannot be confirmed by culture. GPL antibody levels reflect the pulmonary burden of MAC, as assessed from the sputum smear grade and number of involved regions on chest CT.
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Anticuerpos Antibacterianos/sangre , Inmunoglobulina A/sangre , Complejo Mycobacterium avium , Infección por Mycobacterium avium-intracellulare , Esputo/microbiología , Tomografía Computarizada por Rayos X , Anciano , Anciano de 80 o más Años , Femenino , Glucolípidos/sangre , Humanos , Pulmón/diagnóstico por imagen , Pulmón/microbiología , Masculino , Persona de Mediana Edad , Infección por Mycobacterium avium-intracellulare/sangre , Infección por Mycobacterium avium-intracellulare/diagnóstico , Infección por Mycobacterium avium-intracellulare/diagnóstico por imagen , Infección por Mycobacterium avium-intracellulare/microbiologíaRESUMEN
BACKGROUND: Mesenchymal stem cells (MSCs) have been applied to the treatment of various diseases, and MSC administration in marginal donor grafts may help avoid the ischemia-reperfusion injury associated with solid organ transplants. Given the reports of side effects after intravenous MSC administration, local MSC administration to the target organ might be a better approach. We administered adipose tissue-derived MSCs (AT-MSCs) ex vivo to donor rat kidneys obtained after cardiac death (CD). METHODS: Using male Lewis rats (8-10 weeks), and a marginal transplant model of 1hr CD plus 1hr sub-normothermic ET-Kyoto solution preservation were conducted. AT-MSCs obtained from double-reporter (luciferase-LacZ) transgenic Lewis rats were injected either systemically (1.0 × 10(6) cells/0.5 mL) to bilaterally nephrectomized recipient rats that had received a marginal kidney graft (n = 6), or locally via the renal artery (500 µL ET-Kyoto solution containing the same number of AT-MSCs) to marginal kidney grafts, which were then preserved (1 hour; 22°C) before being transplanted into bilaterally nephrectomized recipient rats (n = 8). Serum was collected to assess the therapeutic effects of AT-MSC administration, and the recipients of rats surviving to Day 14 were separately evaluated histopathologically. Follow-up was by in vivo imaging and histological LacZ staining, and tumor formation was evaluated in MSC-injected rats at 3 months. RESULTS: Systemic injection of MSC did not improve recipient survival. In vivo imaging showed MSCs trapped in the lung that later became undetectable. Ex vivo injection of MSCs did show a benefit without adverse effects. At Day 14 after RTx, 75% of the rats in the AT-MSC-injected group (MSC[+]) had survived, whereas 50% of the rats in the AT-MSC-non-injected group (MSC[-]) had died. Renal function in the MSC(+) group was improved compared with that in the MSC(-) group at Day 4. LacZ staining revealed AT-MSCs attached to the renal tubules at 24 hours after RTx that later became undetectable. Histopathologic examination showed little difference in fibrosis between the groups at Day 14. No teratomas or other abnormalities were seen at 3 months.
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Muerte , Riñón/fisiopatología , Trasplante de Células Madre Mesenquimatosas , Tejido Adiposo/citología , Animales , Masculino , Ratas , Ratas Endogámicas Lew , Donantes de TejidosRESUMEN
BACKGROUND: On the basis of our recent findings of oncogenic KRAS-induced interleukin-8 (IL-8) overexpression in non-small cell lung cancer, we assessed the clinicopathological and prognostic significances of IL-8 expression and its relationship to KRAS mutations in lung adenocarcinomas. METHODS: IL-8 expression was examined by quantitative RT-PCR using 136 of surgical specimens from lung adenocarcinoma patients. The association between IL-8 expression, clinicopathological features, KRAS or EGFR mutation status and survival was analysed. RESULTS: IL-8 was highly expressed in tumours from elderly patients or smokers and in tumours with pleural involvement or vascular invasion. In a non-smokers' subgroup, IL-8 level positively correlated with age. IL-8 was highly expressed in tumours with KRAS mutations compared with those with EGFR mutations or wild-type EGFR/KRAS. Lung adenocarcinoma patients with high IL-8 showed significantly shorter disease-free survival (DFS) and overall survival (OS) than those with low IL8. DFS and OS were significantly shorter in the patients with mutant KRAS/high IL-8 than in those with wild-type KRAS/low IL-8. Cox regression analyses demonstrated that elevated IL-8 expression correlated with unfavourable prognosis. CONCLUSIONS: Our findings suggest that IL-8 expression is associated with certain clinicopathological features including age and is a potent prognostic marker in lung adenocarcinoma, especially in oncogenic KRAS-driven adenocarcinoma.
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Adenocarcinoma/genética , Interleucina-8/biosíntesis , Neoplasias Pulmonares/genética , Pronóstico , Proteínas Proto-Oncogénicas/genética , Proteínas ras/genética , Adenocarcinoma/patología , Adenocarcinoma/cirugía , Adenocarcinoma del Pulmón , Adulto , Anciano , Anciano de 80 o más Años , Supervivencia sin Enfermedad , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Interleucina-8/genética , Estimación de Kaplan-Meier , Neoplasias Pulmonares/patología , Neoplasias Pulmonares/cirugía , Masculino , Persona de Mediana Edad , Mutación , Estadificación de Neoplasias , Modelos de Riesgos Proporcionales , Proteínas Proto-Oncogénicas p21(ras)RESUMEN
INTRODUCTION: The solution in which graft tissue is stored (that is, preservation solution) is an important component of liver transplantation technology. Its protective effect is induced by substances in the solution, including radical scavengers, buffers, and energy-giving substances. New preservation solutions have proven to be effective in preventing organ damage during cold ischemia and in extending the time limits for storage. AIM: This study determined the relationship between luminescence intensity and content of adenosine triphosphate (ATP) in liver tissue and proposes a new ex vivo screening system that uses Lewis rats transgenic for luciferase for evaluating the effectiveness of preservation solutions. METHODS: Samples (diameter, 2 mm) of liver were obtained from transgenic rats. The viability of these tissues after storage for as long as 6 hours in University of Wisconsin (UW) solution, extracellular trehalose solution of Kyoto, Euro-Collins (EC) solution, histidine-tryptophan-ketoflutarate solution, low potassium dextran solution, or normal saline was assessed by determining ATP content and luminescence intensity. RESULTS: Luminescence had a linear relationship (R = 0.88) with ATP levels. Regardless of the preservation solution used, the luminescence intensities of the liver tissue chips decreased linearly with time especially through a short span of time (0 to 2 hours; R(2) = 0.58-1.0). The luminescence of liver chip tissues maintained long term (2 to 6 hours) in UW solution tended to be higher than those of tissues stored in other solutions (P < .05; 6 hours). On the basis of luminescence intensity, EC might be preferable to the other solutions tested for ultra-short-term storage (0.5 to 2 hours). CONCLUSION: Our model, which combines the use of the bioimaging system and Lewis rats transgenic for luciferase, effectively assessed the viability of liver tissue samples. We believe that this ex vivo screening system will be an effective tool for evaluating preservation solutions for liver grafts.
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Trasplante de Hígado , Hígado/efectos de los fármacos , Soluciones Preservantes de Órganos/química , Preservación de Órganos/métodos , Adenosina/química , Adenosina Trifosfato/química , Alopurinol/química , Animales , Dextranos/química , Glutatión/química , Histidina/química , Soluciones Hipertónicas/química , Insulina/química , Luciferasas/genética , Luminiscencia , Masculino , Potasio/química , Rafinosa/química , Ratas , Ratas Endogámicas Lew , Ratas Transgénicas , Trehalosa/química , Triptófano/químicaRESUMEN
Retroviral replicating vectors (RRVs) have been shown to achieve efficient tumor transduction and enhanced therapeutic benefit in a wide variety of cancer models. Here we evaluated two different RRVs derived from amphotropic murine leukemia virus (AMLV) and gibbon ape leukemia virus (GALV), in human malignant mesothelioma cells. In vitro, both RRVs expressing the green fluorescent protein gene efficiently replicated in most mesothelioma cell lines tested, but not in normal mesothelial cells. Notably, in ACC-MESO-1 mesothelioma cells that were not permissive for AMLV-RRV, the GALV-RRV could spread efficiently in culture and in mice with subcutaneous xenografts by in vivo fluorescence imaging. Next, GALV-RRV expressing the cytosine deaminase prodrug activator gene showed efficient killing of ACC-MESO-1 cells in a prodrug 5-fluorocytosine dose-dependent manner, compared with AMLV-RRV. GALV-RRV-mediated prodrug activator gene therapy achieved significant inhibition of subcutaneous ACC-MESO-1 tumor growth in nude mice. Quantitative reverse transcription PCR demonstrated that ACC-MESO-1 cells express higher PiT-1 (GALV receptor) and lower PiT-2 (AMLV receptor) compared with normal mesothelial cells and other mesothelioma cells, presumably accounting for the distinctive finding that GALV-RRV replicates much more robustly than AMLV-RRV in these cells. These data indicate the potential utility of GALV-RRV-mediated prodrug activator gene therapy in the treatment of mesothelioma.
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Vectores Genéticos/genética , Virus de la Leucemia del Gibón/genética , Neoplasias Pulmonares/genética , Mesotelioma/genética , Transducción Genética , Animales , Línea Celular Tumoral , Citosina Desaminasa/genética , Citosina Desaminasa/metabolismo , Modelos Animales de Enfermedad , Femenino , Expresión Génica , Orden Génico , Terapia Genética , Vectores Genéticos/administración & dosificación , Humanos , Neoplasias Pulmonares/metabolismo , Mesotelioma/metabolismo , Mesotelioma Maligno , Ratones , Profármacos , Replicación Viral , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Toca 511 (vocimagene amiretrorepvec), an amphotropic retroviral replicating vector (RRV), can successfully and safely deliver a functional, optimized cytosine deaminase (CD) gene to tumors in orthotopic glioma models. This agent, in conjunction with subsequent oral extended-release 5-fluorocytosine (5-FC) (Toca FC), is currently under investigation in patients with recurrent high-grade glioma . Temozolomide (TMZ) with radiation is the most frequently used first-line treatment for patients with glioblastoma, the most common and aggressive form of primary brain cancer in adults. However, subsets of patients with certain genetic alterations do not respond well to TMZ treatment and the overall median survival for patients who respond remains modest, suggesting that combinatorial approaches may be necessary to significantly improve outcomes. We show that in vitro TMZ delays but does not prevent RRV spread, nor interfere with Toca 511+5-FC-mediated cell killing in glioma tumor cells, and in vivo there is no significant hematologic effect from the combination of 5-FC and the clinically relevant dose of TMZ. A synergistic long-term survival advantage is observed in mice bearing an orthotopic TMZ-sensitive glioma after Toca 511 administration followed by coadministration of TMZ and 5-FC. These results provide support for the investigation of this novel combination treatment strategy in patients with newly diagnosed malignant glioma.
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Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Neoplasias Encefálicas/terapia , Citosina Desaminasa/genética , Dacarbazina/análogos & derivados , Flucitosina/farmacología , Glioblastoma/terapia , Animales , Neoplasias Encefálicas/tratamiento farmacológico , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/metabolismo , Citosina Desaminasa/biosíntesis , Citosina Desaminasa/metabolismo , Dacarbazina/administración & dosificación , Dacarbazina/farmacología , Sinergismo Farmacológico , Femenino , Flucitosina/administración & dosificación , Flucitosina/farmacocinética , Expresión Génica , Técnicas de Transferencia de Gen , Vectores Genéticos/genética , Glioblastoma/tratamiento farmacológico , Glioblastoma/genética , Glioblastoma/metabolismo , Ratones , Ratones Desnudos , Retroviridae/genética , Temozolomida , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
BACKGROUND: Segmental intestinal transplantations from living, genetically related donors provide advantages compared with those from cadaveric subjects. However, successful preservation during ischemic cold storage is critical for living donor grafts. Thus, the development of preservation solutions that maintain graft viability is essential for success. Herein we have reported application of a cell-based viability assay in multiwell plates to assess the effectiveness of various solutions to preserve intestinal grafts. METHODS: Freshly isolated intestinal chips from luciferase transgenic rats were placed in 96-well tissue culture plates for incubation at 4°C for 24 hours in various preservation solutions: ET-Kyoto (ET-K), University of Wisconsin (UW) solution, Euro-Collins (EC) solution, histidine-tryptophan-ketoglutarate (HTK) solution, lactated Ringer's (LR) solution, or saline. RESULTS: As indicated by a higher level of luminescence, intestinal chips preserved in UW, HTK, or ET-K solution contained more viable cells, than those preserved in EC, LR, or saline solution. After exposure to the preservation solutions for 1 hour, the mucosal layer chips showed lower cell viability than the muscle layer chips. CONCLUSION: Our data demonstrated that ET-K and UW solutions used together with intestinal chips of Luciferase transgenic rat and in vivo imaging provided optimal viability during ischemic cold storage prior to transplantation. Further development of preservation conditions to minimize the loss of viability of intestinal grafts before clinical transplantation is essential to improve outcomes.
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Ensayos Analíticos de Alto Rendimiento/métodos , Intestino Delgado/efectos de los fármacos , Intestino Delgado/trasplante , Soluciones Preservantes de Órganos/farmacología , Preservación de Órganos/métodos , Adenosina/farmacología , Adenosina Trifosfato/metabolismo , Alopurinol/farmacología , Animales , Supervivencia Celular/efectos de los fármacos , Isquemia Fría/efectos adversos , Gluconatos/farmacología , Glucosa/farmacología , Glutatión/farmacología , Derivados de Hidroxietil Almidón/farmacología , Soluciones Hipertónicas/farmacología , Insulina/farmacología , Intestino Delgado/metabolismo , Intestino Delgado/patología , Soluciones Isotónicas/farmacología , Luciferasas/biosíntesis , Luciferasas/genética , Mediciones Luminiscentes , Manitol/farmacología , Fosfatos/farmacología , Cloruro de Potasio/farmacología , Procaína/farmacología , Rafinosa/farmacología , Ratas , Ratas Transgénicas , Lactato de Ringer , Cloruro de Sodio/farmacología , Espectrometría de Fluorescencia , Factores de Tiempo , Técnicas de Cultivo de Tejidos , Trehalosa/farmacologíaRESUMEN
We have recently developed a novel and highly efficient strategy that exclusively uses the purine analog 6-thioguanine (6TG) for both pretransplantation conditioning and post-transplantation chemoselection of hypoxanthine-guanine phosphoribosyltransferase (HPRT)-deficient bone marrow (BM). In a mouse BM transplantation model, combined 6TG preconditioning and in vivo chemoselection consistently achieved >95% engraftment of HPRT-deficient donor BM and long-term reconstitution of histologically and immunophenotypically normal hematopoiesis in both primary and secondary recipients, without significant toxicity and in the absence of any other cytotoxic conditioning regimen. To translate this strategy for combined 6TG conditioning and chemoselection into a clinically feasible approach, it is necessary to develop methods for genetic modification of normal hematopoietic stem cells (HSC) to render them HPRT-deficient and thus 6TG-resistant. Here we investigated a strategy to reduce HPRT expression and thereby confer protection against 6TG myelotoxicity to primary murine BM cells by RNA interference (RNAi). Accordingly, we constructed and validated a lentiviral gene transfer vector expressing short-hairpin RNA (shRNA) that targets the murine HPRT gene. Our results showed that lentiviral vector-mediated delivery of HPRT-targeted shRNA could achieve effective and long-term reduction of HPRT expression. Furthermore, in both an established murine cell line as well as in primary murine BM cells, lentiviral transduction with HPRT-targeted shRNA was associated with enhanced resistance to 6TG cytotoxicity in vitro. Hence this represents a translationally feasible method to genetically engineer HSC for implementation of 6TG-mediated preconditioning and in vivo chemoselection.
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Médula Ósea/enzimología , Vectores Genéticos , Hipoxantina Fosforribosiltransferasa/genética , Lentivirus/genética , ARN Interferente Pequeño/genética , Tioguanina/farmacología , Animales , Trasplante de Médula Ósea , Línea Celular Tumoral , Ratones , Interferencia de ARNRESUMEN
In the present study, we compared the therapeutic effect of tumor-selective retroviral replicating vectors (RRV) expressing the yeast cytosine deaminase (CD) delivered by convection-enhanced delivery (CED) or simple injection, followed by systemic administration of the pro-drug, 5-fluorocytosine (5-FC). Treatment with RRV-CD and systemic 5-FC significantly increased survival in rodent U87MG glioma model in comparison with controls (P<0.01). Interestingly, CED of RRV-CD followed by 5-FC further enhanced survival in this animal model in comparison with intra-tumoral injection of RRV-CD, followed by systemic 5-FC (P<0.05). High expression levels of Ki-67 were found in untreated tumors compared with treated. Untreated tumors were also much larger than treated. CED resulted in excellent distribution of RRV while only partial distribution of RRV was obtained after injection. Furthermore, RRV-CD and CD were also found in tumors from treated rats at study end points. These results demonstrated that RRV vectors may efficiently transduce and stably propagate in malignant human glioma, thereby achieving a significant in situ amplification effect after initial administration. We conclude that delivery of RRV into the glioma by CED provides much wider vector distribution than simple injection, and this correlated with better therapeutic outcomes.
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Neoplasias Encefálicas/tratamiento farmacológico , Citosina Desaminasa/administración & dosificación , Flucitosina/administración & dosificación , Glioma/tratamiento farmacológico , Animales , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/patología , Convección , Citosina Desaminasa/genética , Sistemas de Liberación de Medicamentos , Terapia Genética , Vectores Genéticos/administración & dosificación , Glioma/genética , Glioma/patología , Humanos , Antígeno Ki-67/biosíntesis , Ratas , RetroviridaeRESUMEN
We provide an overview of the latest developments in cancer gene therapy--from the bench to early-stage clinical trials. We describe the most recent work of worldwide teams including experienced scientists and clinicians, reflecting the recent emergence of gene therapy from the 'Valley of Death'. The treatment efficacy of clinical gene therapy has now been shown in a number of diseases including cancer and we are observing a renewed interest by big pharmaceutical and biotechnology companies most obviously demonstrated by Amgen's acquisition of Biovex for up to USD$1 billion. There is an opportunity to be cautiously hopeful regarding the future of gene therapy in the clinic and we review here some of the most recent progress in the field.
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Tratamiento Basado en Trasplante de Células y Tejidos , Terapia Genética , Neoplasias/genética , Neoplasias/terapia , Animales , Biomarcadores de Tumor/genética , Ensayos Clínicos Fase I como Asunto , Ensayos Clínicos Fase II como Asunto , Técnicas de Transferencia de Gen , Vectores Genéticos , Humanos , Inmunoterapia , Terapia Molecular Dirigida , Medicina de Precisión , Interferencia de ARNRESUMEN
PURPOSE: To assess the prognostic value of the Ocular Trauma Score (OTS) in patients with eye injury treated in a University Hospital in South America. METHODS: One hundred and ninety subjects who were victims of eye trauma with different degrees of severity were evaluated and treated accordingly. Initial OTS categories were calculated for each patient and compared to the final visual result by transforming the achieved best-corrected visual acuity (BCVA) after treatment into OTS values. RESULTS: OTS visual acuity showed good correlation with the final BCVA after treatment [Spearman's ρ = 0.857, 95 % confidence interval (CI) 0.813-0.891, P < 0.0001). No difference between the predicted BCVA and achieved BCVA was noticed (P < 0.0001, Chi-square test). CONCLUSION: The OTS provided useful information on outcomes treatment after eye trauma and can be used as a prognostic model to predict vision survival in injured adult patients from a developing country.
RESUMEN
Replication-competent retrovirus (RCR) vectors have been shown to achieve significantly enhanced tumor transduction efficiency and therapeutic efficacy in various cancer models. In the present study, we investigated RCR vector-mediated prodrug activator gene therapy for the treatment of malignant mesothelioma, a highly aggressive tumor with poor prognosis. RCR-GFP vector expressing the green fluorescent protein marker gene successfully infected and efficiently replicated in human malignant mesothelioma cell lines, as compared with non-malignant mesothelial cells in vitro. In mice with pre-established subcutaneous tumor xenografts, RCR-GFP vector showed robust spread throughout entire tumor masses after intratumoral administration. Next, RCR-cytosine deaminase (RCR-CD), expressing the yeast CD prodrug activator gene, showed efficient transmission of the prodrug activator gene associated with replicative spread of the virus, resulting in efficient killing of malignant mesothelioma cells in a prodrug 5-fluorocytosine (5FC)-dose dependent manner in vitro. After a single intratumoral injection of RCR-CD followed by intraperitoneal administration of 5FC, RCR vector-mediated prodrug activator gene therapy achieved significant inhibition of subcutaneous tumor growth, and significantly prolonged survival in the disseminated peritoneal model of malignant mesothelioma. These data indicate the potential utility of RCR vector-mediated prodrug activator gene therapy in the treatment of malignant mesothelioma.
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Terapia Genética/métodos , Mesotelioma/terapia , Mesotelioma/virología , Viroterapia Oncolítica/métodos , Retroviridae/genética , Adulto , Animales , Línea Celular Tumoral , Citosina Desaminasa/genética , Citosina Desaminasa/metabolismo , Modelos Animales de Enfermedad , Femenino , Flucitosina/administración & dosificación , Flucitosina/farmacocinética , Humanos , Masculino , Mesotelioma/genética , Mesotelioma/metabolismo , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Profármacos/administración & dosificación , Profármacos/farmacocinética , Retroviridae/fisiología , Análisis de Supervivencia , Transducción Genética , Replicación ViralRESUMEN
Development of lentiviral vectors (LVs) in the field of immunotherapy and immune regeneration will strongly rely on biosafety of the gene transfer. We demonstrated previously the feasibility of ex vivo genetic programming of mouse bone marrow precursors with LVs encoding granulocyte macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4), which induced autonomous differentiation of long-lived dendritic cells (DCs), referred to as self-differentiated myeloid-derived antigen-presenting-cells reactive against tumors (SMART-DCs). Here, LV biosafety was enhanced by using a DC-restricted and physiological promoter, the major histocompatibility complex (MHC) II promoter, and including co-expression of the herpes simplex virus-thymidine kinase (sr39HSV-TK) conditional suicide gene. Tricistronic vectors co-expressing sr39HSV-TK, GM-CSF and IL-4 transcriptionally regulated by the MHCII promoter or the ubiquitous cytomegalovirus (CMV) promoter were compared. Despite the different gene transfer effects, such as the kinetics, levels of transgene expression and persistency of integrated vector copies, both vectors induced highly viable SMART-DCs, which persisted for at least 70 days in vivo and could be ablated with the pro-drug Ganciclovir (GCV). SMART-DCs co-expressing the tyrosine-related protein 2 melanoma antigen administered subcutaneously generated antigen-specific, anti-melanoma protective and therapeutic responses in the mouse B16 melanoma model. GCV administration after immunotherapy did not abrogate DC vaccination efficacy. This demonstrates proof-of-principle of genetically programmed DCs that can be ablated pharmacologically.
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Diferenciación Celular/genética , Células Dendríticas/inmunología , Vectores Genéticos , Lentivirus/genética , Melanoma Experimental/terapia , Animales , Movimiento Celular , Supervivencia Celular , Ganciclovir/farmacología , Genes MHC Clase II , Genes Transgénicos Suicidas , Interleucina-4 , Ratones , Ratones Endogámicos C57BL , Simplexvirus/genética , Timidina Quinasa/genética , VacunaciónRESUMEN
Gene transfer of the Escherichia coli purine nucleoside phosphorylase (PNP) results in potent cytotoxicity after administration of the prodrug fludarabine phosphate (F-araAMP). Here, we have tested whether application of this strategy in the context of replication-competent retrovirus (RCR) vectors, which can achieve highly efficient tumor-restricted transduction as well as persistent expression of transgenes, would result in effective tumor inhibition, or, alternatively, would adversely affect viral replication. We found that RCR vectors could achieve high levels of PNP expression concomitant with the efficiency of their replicative spread, with significant cell killing activity in vitro and potent therapeutic effects in vivo. In U-87 xenograft models, replicative spread of the vector resulted in progressive transmission of the PNP transgene, as evidenced by increasing PNP enzyme activity with time after vector inoculation. On F-araAMP administration, high efficiency gene transfer of PNP by the RCR vector resulted in significant suppression of tumor growth and extended survival time. As the RCR mediates stable integration of the PNP gene and continuous expression, an additional round of F-araAMP administration resulted in further survival benefit. RCR-mediated PNP suicide gene therapy thus represents a highly efficient form of intracellular chemotherapy, and may achieve effective antitumor activity with less systemic toxicity.
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Escherichia coli/enzimología , Vectores Genéticos , Glioma/terapia , Profármacos/farmacología , Purina-Nucleósido Fosforilasa/genética , Retroviridae/genética , Fosfato de Vidarabina/análogos & derivados , Animales , Antimetabolitos Antineoplásicos/farmacología , Terapia Genética , Glioma/genética , Glioma/virología , Proteínas Fluorescentes Verdes/metabolismo , Humanos , Inyecciones Intralesiones , Inyecciones Subcutáneas , Ratones , Ratones Desnudos , Fosfato de Vidarabina/farmacologíaRESUMEN
We have developed unique replication-competent retroviral (RCR) vectors based on murine leukemia virus that provide improved efficiency of viral delivery, allow for long-term transgene expression and demonstrate an intrinsic selectivity for transduction of rapidly dividing tumor cells. The purpose of this study was to evaluate the in vivo transduction efficiency and the therapeutic efficacy of the RCR vector mediated delivery of Escherichia coli purine nucleoside phosphorylase (PNP) in combination with fludarabine phosphate for bladder cancer. We constructed vectors containing green fluorescent protein (GFP) gene (ACE)-GFP) or PNP gene (ACE-PNP). KU-19-19 bladder tumors exhibited 28.3+/-16.1, 46.6+/-5.8 and 93.7+/-7.8% of GFP expression on 14, 18 and 26 days after intratumoral injection of ACE-GFP, respectively. GFP expression could not be observed in normal tissues surrounding the injected tumors. No detectable polymerase chain reaction products of GFP gene could be observed in any distant organs. Intratumoral injection of ACE-PNP, followed by systemically administered fludarabine phosphate, significantly inhibited the growth of pre-established KU-19-19 tumors. Our results indicate that RCR vectors are a potentially efficient gene delivery method and that the RCR vector mediated PNP gene transfer and fludarabine phosphate treatment might be a novel and potentially therapeutic modality for bladder cancer.
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Antimetabolitos Antineoplásicos/metabolismo , Escherichia coli/enzimología , Virus de la Leucemia Murina/genética , Profármacos/metabolismo , Purina-Nucleósido Fosforilasa/genética , Neoplasias de la Vejiga Urinaria/terapia , Fosfato de Vidarabina/análogos & derivados , Animales , Terapia Combinada , Replicación del ADN , Técnicas de Transferencia de Gen , Terapia Genética , Vectores Genéticos , Proteínas Fluorescentes Verdes/genética , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Purina-Nucleósido Fosforilasa/uso terapéutico , Transducción Genética , Trasplante Heterólogo , Células Tumorales Cultivadas , Neoplasias de la Vejiga Urinaria/genética , Neoplasias de la Vejiga Urinaria/patología , Fosfato de Vidarabina/metabolismoAsunto(s)
Anticuerpos Monoclonales/farmacología , Leucemia/tratamiento farmacológico , Linfocitos T/efectos de los fármacos , Animales , Anticuerpos Monoclonales Humanizados , Antígenos CD/efectos de los fármacos , Antígenos CD/inmunología , Antígenos de Diferenciación/efectos de los fármacos , Antígenos de Diferenciación/inmunología , Antígeno CTLA-4 , HumanosRESUMEN
Transplantation of many tissues requires histocompatibility matching of human leukocyte antigens (HLA) to prevent graft rejection, to reduce the level of immunosuppression needed to maintain graft survival, and to minimize the risk of graft-versus-host disease, particularly in the case of bone marrow transplantation. However, recent advances in fields of gene delivery and genetic regulation technologies have opened the possibility of engineering grafts that display reduced levels of HLA expression. Suppression of HLA expression could help to overcome the limitations imposed by extensive HLA polymorphisms that restrict the availability of suitable donors, necessitate the maintenance of large donor registries, and complicate the logistics of procuring and delivering matched tissues and organs to the recipient. Accordingly, we investigated whether knockdown of HLA by RNA interference (RNAi), a ubiquitous regulatory system that can efficiently and selectively inhibit the expression of specific gene products, would enable allogeneic cells to evade immune recognition. For efficient and stable delivery of short hairpin-type RNAi constructs (shRNA), we employed lentivirus-based gene transfer vectors, which provide a delivery system that can achieve integration into genomic DNA, thereby permanently modifying transduced graft cells. Our results show that lentivirus-mediated delivery of shRNA targeting pan-Class I and allele-specific HLA can achieve efficient and dose-dependent reduction in surface expression of HLA in human cells, associated with enhanced resistance to alloreactive T lymphocyte-mediated cytotoxicity, while avoiding MHC-non-restricted killing. We hypothesize that RNAi-induced silencing of HLA expression has the potential to create histocompatibility-enhanced, and, eventually, perhaps "universally" compatible cellular grafts.
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Antígenos de Histocompatibilidad Clase I/genética , Antígenos de Histocompatibilidad Clase I/inmunología , Linfocitos T/inmunología , Secuencia de Bases , Línea Celular , Citotoxicidad Inmunológica , Cartilla de ADN , Silenciador del Gen , Técnicas de Transferencia de Gen , Vectores Genéticos , VIH/inmunología , Humanos , Interferón gamma/inmunología , Riñón , Lentivirus , Interferencia de ARNRESUMEN
Squamous cell carcinoma antigens SCCA1 and SCCA2 are highly homologous serine proteinase inhibitors which have been widely utilized as serological markers for squamous cell cancers, but it has recently been demonstrated that only SCCA2 is truly specific for certain forms of lung cancer. Using a construct containing the 5'-flanking region of the SCCA2 gene between -460 and +0 bp and the luciferase reporter gene, SCCA2 promoter activity was detected in SCCA2-producing SCC cell lines (LK-2, LC-1), but not in SCCA2-nonproducing lung adenocarcinoma cell lines (A549, ABC-1, and RERF-LC-MS) or normal cells (WI-38, SAEC, and NHEK-Adult). Infection with a recombinant adenovirus vector, Ad-SCCA2-DsRed, resulted in cell-specific expression of the SCCA2 promoter-driven DsRed marker gene only in LK-2 and LC-1 cells. The same strategy was used for SCCA2-driven expression of a proapoptotic gene, (KLAKLAK)2, which can cause mitochondrial disruption by triggering mitochondrial permeabilization and swelling, resulting in the release of cytochrome c and induction of apoptosis. Infection with Ad-SCCA2-KLAKLAK2 specifically reduced the growth of the two human lung SCC cell lines compared to the SCCA2 nonproducing cell lines both in vitro and in vivo, suggesting that the SCCA2 promoter had a tumor-specific effect. These results suggest that transduction of SCCA2 promoter-controlled suicide genes by adenoviral vectors can confer transcriptionally targeted cytotoxicity in SCCA2-producing lung SCC cells, and represents a novel strategy for gene transfer specifically targeted to SCC in the lung.