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1.
Clin Cancer Res ; 24(18): 4388-4398, 2018 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-29773661

RESUMO

Purpose: Peritoneal carcinomatosis is common in advanced tumor stages or disease recurrence arising from gastrointestinal cancers, gynecologic malignancies, or primary peritoneal carcinoma. Because current therapies are mostly ineffective, new therapeutic approaches are needed. Here, we report on a phase I study designed to assess safety, MTD, and antitumor activity of intraperitoneal administration of oncolytic vaccinia virus GL-ONC1 in advanced stage peritoneal carcinomatosis patients.Patients and Methods: GL-ONC1 was administered intraperitoneally every 4 weeks for up to four cycles at three different dose levels (107-109 pfu) following a standard 3+3 dose escalation design. GL-ONC1 was infused via an indwelling catheter that enabled repetitive analyses of peritoneal fluid biopsies. The primary study objective was safety of GL-ONC1 according to Common Terminology Criteria for Adverse Events, version 4.0 (CTCAEv4.0).Results: Patients with advanced-stage peritoneal carcinomatosis (n = 7) or advanced peritoneal mesothelioma (n = 2) received 24 doses of GL-ONC1. Adverse events were limited to grades 1-3, including transient flu-like symptoms and increased abdominal pain, resulting from treatment-induced peritonitis. No DLT was reported, and the MTD was not reached. Furthermore, no signs of viral shedding were observed. Importantly, in 8 of 9 study patients, effective intraperitoneal infections, in-patient replication of GL-ONC1, and subsequent oncolysis were demonstrated in cycle 1. All patients developed neutralizing activities against GL-ONC1.Conclusions: GL-ONC1 was well tolerated when administered into the peritoneal cavity of patients with advanced stage peritoneal carcinomatosis. Efficient tumor cell infection, in-patient virus replication, and oncolysis were limited to treatment cycle 1 (ClinicalTrials.gov number, NCT01443260). Clin Cancer Res; 24(18); 4388-98. ©2018 AACR.


Assuntos
Neoplasias Pulmonares/terapia , Mesotelioma/terapia , Terapia Viral Oncolítica/efeitos adversos , Neoplasias Peritoneais/terapia , Vírus Vaccinia/genética , Adulto , Idoso , Líquido Ascítico/virologia , Linhagem Celular Tumoral , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos/classificação , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos/patologia , Feminino , Humanos , Injeções Intraperitoneais , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/virologia , Masculino , Mesotelioma/genética , Mesotelioma/patologia , Mesotelioma/virologia , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/genética , Recidiva Local de Neoplasia/patologia , Recidiva Local de Neoplasia/terapia , Recidiva Local de Neoplasia/virologia , Estadiamento de Neoplasias , Vírus Oncolíticos/genética , Neoplasias Peritoneais/genética , Neoplasias Peritoneais/patologia , Neoplasias Peritoneais/virologia , Replicação Viral/genética
2.
Virol J ; 14(1): 219, 2017 11 10.
Artigo em Inglês | MEDLINE | ID: mdl-29126418

RESUMO

BACKGROUND: Genetically modified adenoviruses (Ad) with preferential replications in tumor cells have been examined for a possible clinical applicability as an anti-cancer agent. A simple method to detect viral and cellular proteins is valuable to monitor the viral infections and to predict the Ad-mediated cytotoxicity. METHODS: We used type 5 Ad in which the expression of E1A gene was activated by 5'-regulatory sequences of genes that were augmented in the expression in human tumors. The Ad were further modified to have the fiber-knob region replaced with that derived from type 35 Ad. We infected human mesothelioma cells with the fiber-replaced Ad, and sequentially examined cytotoxic processes together with an expression level of the viral E1A, hexon, and cellular cleaved caspase-3 with image cytometric and Western blot analyses. RESULTS: The replication-competent Ad produced cytotoxicity on mesothelioma cells. The infected cells expressed E1A and hexon 24 h after the infection and then showed cleavage of caspase-3, all of which were detected with image cytometry and Western blot analysis. Image cytometry furthermore demonstrated that increased Ad doses did not enhance an expression level of E1A and hexon in an individual cell and that caspase-3-cleaved cells were found more frequently in hexon-positive cells than in E1A-positive cells. Image cytometry thus detected these molecular changes in a sensitive manner and at a single cell level. We also showed that an image cytometric technique detected expression changes of other host cell proteins, cyclin-E and phosphorylated histone H3 at a single cell level. CONCLUSIONS: Image cytometry is a concise procedure to detect expression changes of Ad and host cell proteins at a single cell level, and is useful to analyze molecular events after the infection.


Assuntos
Vetores Genéticos/fisiologia , Citometria por Imagem , Neoplasias Pulmonares/virologia , Mesotelioma/virologia , Terapia Viral Oncolítica/métodos , Adenoviridae/genética , Proteínas E1A de Adenovirus/metabolismo , Proteínas do Capsídeo/metabolismo , Caspase 3/metabolismo , Morte Celular , Linhagem Celular Tumoral , Vetores Genéticos/genética , Células HEK293 , Interações Hospedeiro-Patógeno , Humanos , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Mesotelioma/metabolismo , Mesotelioma/patologia , Análise de Célula Única , Replicação Viral
3.
Toxicol In Vitro ; 41: 49-55, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28223140

RESUMO

Besides its well-recognized antiviral activity, Cidofovir (CDV) has been shown to exert anticancer properties both within in vitro and in vivo models. The aim of this study was to evaluate the effects of CDV on still unexplored cultured cancer cells from human mesothelioma as well as breast, colon, liver, lung, prostate, and thyroid carcinomas. Overall, a dose- and time-dependent inhibition of cell viability was observed after CDV exposure. To clarify the mechanisms underlying CDV action, apoptotic cell death was investigated in two infected cell lines [Ist-Mes1 and Ist-Mes2 mesothelioma cells (SV40+)] and in two uninfected cell lines (NCI-H2425 mesothelioma cells and FTC-133 thyroid cancer cells), which resulted the most sensitive to CDV treatment. Reduced expression of procaspase-3 and increased expression of PARP p85 fragment were observed in both infected and uninfected mesothelioma cells, indicating apoptosis induction by CDV in a virus-independent manner. Similarly, the increase of the pro-apoptotic proteins p53, cytochrome c and caspase-3, the decrease of the survival protein Bcl-x, and the increment of Bax/Bcl-2 ratio revealed the occurrence of apoptosis in CDV-treated FTC-133. The presence of nuclear DNA fragmentation confirmed apoptotic cell death by CDV. Overall, our findings warrant further investigations to explore the therapeutic potential of CDV for human mesothelioma and follicular thyroid carcinoma.


Assuntos
Antineoplásicos/farmacologia , Antivirais/farmacologia , Citosina/análogos & derivados , Organofosfonatos/farmacologia , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Cidofovir , Citosina/farmacologia , Fragmentação do DNA , Feminino , Humanos , Mesotelioma/tratamento farmacológico , Mesotelioma/virologia , Infecções por Papillomavirus , Infecções por Polyomavirus , Neoplasias da Glândula Tireoide/tratamento farmacológico , Infecções Tumorais por Vírus , Neoplasias do Colo do Útero/tratamento farmacológico , Neoplasias do Colo do Útero/virologia
4.
Cancer Sci ; 108(1): 116-123, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-27783867

RESUMO

Malignant mesothelioma (MM) incidence is increasing drastically worldwide as an occupational disease resulting from asbestos exposure. However, no curative treatment for MM of advanced stage is available. Thus, new therapeutic approaches for MM are required. Because malignant pleural mesothelioma (MPM) cells spread along the pleural surface in most patients, MPM can be targeted using intrapleural therapeutic approaches. In this study, we investigated the effectiveness of the intrapleural instillation of a replication-competent adenovirus as an oncolytic agent against MPM. We constructed a vascular endothelial growth factor promoter-based conditionally replicative adenovirus (VEGF-CRAd) that replicates exclusively in VEGF-expressing cells. All of the MM cell lines that we tested expressed VEGF mRNA, and VEGF-CRAd selectively replicated in these MM cells and exerted a direct concentration-dependent oncolytic effect in vitro. Furthermore, our in vivo studies showed that pre-infection of MM cells with VEGF-CRAd potently suppressed MPM tumor formation in nude mice, and that intrapleural instillation of VEGF-CRAd prolonged the survival time of tumor-bearing mice. Our results indicate that VEGF-CRAd exerts an oncolytic effect on MM cells and that intrapleural instillation of VEGF-CRAd is safe and might represent a promising therapeutic strategy for MPM.


Assuntos
Adenoviridae/crescimento & desenvolvimento , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/terapia , Mesotelioma/patologia , Mesotelioma/terapia , Terapia Viral Oncolítica , Neoplasias Pleurais/terapia , Regiões Promotoras Genéticas/genética , Fator A de Crescimento do Endotélio Vascular/genética , Replicação Viral/genética , Adenoviridae/genética , Animais , Morte Celular , Linhagem Celular Tumoral , Feminino , Humanos , Neoplasias Pulmonares/virologia , Mesotelioma/virologia , Camundongos , Camundongos Nus , Neoplasias Pleurais/patologia , Neoplasias Pleurais/virologia , Transgenes/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto
5.
Int J Cancer ; 139(8): 1883-93, 2016 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-27287512

RESUMO

Malignant mesothelioma (MM) is a rare cancer type caused mainly by asbestos exposure. The median overall survival time of a mesothelioma cancer patient is less than 1-year from diagnosis. Currently there are no curative treatment modalities for malignant mesothelioma, however treatments such as surgery, chemotherapy and radiotherapy can help to improve patient prognosis and increase life expectancy. Pemetrexed-Cisplatin is the only standard of care (SoC) chemotherapy for malignant mesothelioma, but the median PFS/OS (progression-free survival/overall survival) from the initiation of treatment is only up to 12 months. Therefore, new treatment strategies against malignant mesothelioma are in high demand. ONCOS-102 is a dual targeting, chimeric oncolytic adenovirus, coding for human GM-CSF. The safety and immune activating properties of ONCOS-102 have already been assessed in phase 1 study (NCT01598129). In this preclinical study, we evaluated the antineoplastic activity of combination treatment with SoC chemotherapy (Pemetrexed, Cisplatin, Carboplatin) and ONCOS-102 in xenograft BALB/c model of human malignant mesothelioma. We demonstrated that ONCOS-102 is able to induce immunogenic cell death of human mesothelioma cell lines in vitro and showed anti-tumor activity in the treatment of refractory H226 malignant pleural mesothelioma (MPM) xenograft model. While chemotherapy alone showed no anti-tumor activity in the mesothelioma mouse model, ONCOS-102 was able to slow down tumor growth. Interestingly, a synergistic anti-tumor effect was seen when ONCOS-102 was combined with chemotherapy regimens. These findings give a rationale for the clinical testing of ONCOS-102 in combination with first-line chemotherapy in patients suffering from malignant mesothelioma.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Neoplasias Pulmonares/terapia , Mesotelioma/terapia , Terapia Viral Oncolítica/métodos , Adenoviridae/genética , Adenoviridae/imunologia , Adenoviridae/fisiologia , Animais , Carboplatina/administração & dosagem , Linhagem Celular Tumoral , Terapia Combinada , Fator Estimulador de Colônias de Granulócitos e Macrófagos/administração & dosagem , Fator Estimulador de Colônias de Granulócitos e Macrófagos/genética , Fator Estimulador de Colônias de Granulócitos e Macrófagos/imunologia , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/virologia , Mesotelioma/tratamento farmacológico , Mesotelioma/imunologia , Mesotelioma/virologia , Camundongos , Camundongos Endogâmicos BALB C , Pemetrexede/administração & dosagem , Replicação Viral , Ensaios Antitumorais Modelo de Xenoenxerto
6.
Oncotarget ; 6(42): 44892-904, 2015 Dec 29.
Artigo em Inglês | MEDLINE | ID: mdl-26539644

RESUMO

Attenuated measles virus (MV) is currently being evaluated as an oncolytic virus in clinical trials and could represent a new therapeutic approach for malignant pleural mesothelioma (MPM). Herein, we screened the sensitivity to MV infection and replication of twenty-two human MPM cell lines and some healthy primary cells. We show that MV replicates in fifteen of the twenty-two MPM cell lines. Despite overexpression of CD46 by a majority of MPM cell lines compared to healthy cells, we found that the sensitivity to MV replication did not correlate with this overexpression. We then evaluated the antiviral type I interferon (IFN) responses of MPM cell lines and healthy cells. We found that healthy cells and the seven insensitive MPM cell lines developed a type I IFN response in presence of the virus, thereby inhibiting replication. In contrast, eleven of the fifteen sensitive MPM cell lines were unable to develop a complete type I IFN response in presence of MV. Finally, we show that addition of type I IFN onto MV sensitive tumor cell lines inhibits replication. These results demonstrate that defects in type I IFN response are frequent in MPM and that MV takes advantage of these defects to exert oncolytic activity.


Assuntos
Interferon Tipo I/metabolismo , Vírus do Sarampo/crescimento & desenvolvimento , Mesotelioma/terapia , Terapia Viral Oncolítica/métodos , Vírus Oncolíticos/crescimento & desenvolvimento , Neoplasias Pleurais/terapia , Replicação Viral , Antígenos CD/metabolismo , Moléculas de Adesão Celular/metabolismo , Linhagem Celular Tumoral , Interações Hospedeiro-Patógeno , Humanos , Interferon Tipo I/imunologia , Vírus do Sarampo/imunologia , Vírus do Sarampo/metabolismo , Proteína Cofatora de Membrana/metabolismo , Mesotelioma/imunologia , Mesotelioma/metabolismo , Mesotelioma/virologia , Vírus Oncolíticos/imunologia , Vírus Oncolíticos/metabolismo , Neoplasias Pleurais/imunologia , Neoplasias Pleurais/metabolismo , Neoplasias Pleurais/virologia , Receptores de Superfície Celular/metabolismo , Transdução de Sinais , Membro 1 da Família de Moléculas de Sinalização da Ativação Linfocitária , Fatores de Tempo
7.
Tumour Biol ; 35(2): 889-901, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24081673

RESUMO

The malignant mesothelioma is an aggressive form of cancer with a mean survival rate of less than a year. Moreover, environmental exposure to minerals is an important factor in the development of malignant mesothelioma (MM), especially the mineral asbestos, which has a well-documented role in MM, and more recently, the mineral erionite has been proven to be a strong carcinogenic inducer of MM. In addition, the virus simian virus 40 has been implicated as a co-carcinogenic player in MM. However, the molecular mechanisms involved in the pathogenesis of this cancer are still not fully understood. Indeed, it is known that several genes are altered or mutated in MM, among those are p16(INK4A), p14(ARF), and neurofibromatosis type II. Furthermore, TP53 has been reported to be mutated in the majority of the cancers; however, in MM, it is very uncommon mutations in this gene. Also, the PTEN gene has been shown to play an important role in endometrial cancer and glioblastoma, although the role of PTEN in MM has yet to be established. Taken altogether, this review focuses on the historical aspects, molecular mechanisms, interaction with other genes and proteins, and the role of these genes in MM. Lastly, this review questions the cancer theory of the two hits because the functions of both PTEN and TP53 are not fully explained by this theory.


Assuntos
Asbestos/toxicidade , Neoplasias Pulmonares/genética , Proteínas de Membrana/genética , Mesotelioma/genética , PTEN Fosfo-Hidrolase/genética , Proteína Supressora de Tumor p53/genética , Carcinógenos/toxicidade , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Neoplasias Pulmonares/induzido quimicamente , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/virologia , Proteínas de Membrana/metabolismo , Mesotelioma/induzido quimicamente , Mesotelioma/patologia , Mesotelioma/virologia , PTEN Fosfo-Hidrolase/metabolismo , Vírus 40 dos Símios/patogenicidade , Proteína Supressora de Tumor p53/metabolismo
8.
Am J Ind Med ; 56(10): 1221-5, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23828611

RESUMO

BACKGROUND: Diffuse malignant mesothelioma (DMM) is increasing in incidence on a worldwide basis and is linked to exposure to asbestos. Simian virus 40 (SV40), a DNA virus, was introduced inadvertently to human populations through contaminated polio vaccine during the years 1956-1963. It has been associated with various types of malignancy in animal experiments. There have been suggestions that SV40 might play a role in the pathogenesis of DMM. OBJECTIVE: To evaluate the association between SV40 and DMM in Iranian patients. METHOD: In a case-control study between the years 2007-2008, isolated DNA from 60 paraffin blocks of patients with DMM and 60 controls was assessed to detect three human polyomaviruses (JCV, BKV, and SV40) using three different sets of primers by multiplex nested PCR analysis. We related the patients with diffuse malignant mesothelioma to possible sites of exposure to asbestos. RESULTS: None of the DMMs nor any patient in the control group had SV40 genome on polymerase chain reaction (PCR). All of the cases were SV40 T antigen negative. CONCLUSION: This study suggests that DMM is independent of SV40 infection in Iran.


Assuntos
Mesotelioma/virologia , Neoplasias Pleurais/virologia , Infecções por Polyomavirus/virologia , Infecções Tumorais por Vírus/virologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Feminino , Humanos , Irã (Geográfico)/epidemiologia , Masculino , Mesotelioma/epidemiologia , Pessoa de Meia-Idade , Neoplasias Pleurais/epidemiologia , Reação em Cadeia da Polimerase , Infecções por Polyomavirus/epidemiologia , Vírus 40 dos Símios/genética , Infecções Tumorais por Vírus/epidemiologia
9.
Anticancer Res ; 32(9): 3743-7, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22993314

RESUMO

BACKGROUND: Adenovirus vectors have been utilized for cancer gene therapies. The present study examined the oncolytic effects of adenovirus type 5 (Ad5) and fiber-substituted conditionally replicating adenovirus (CRAD) Ad5/F35 vectors on the human malignant mesothelioma cells MSTO-211H, NCI-H28, NCI-H2052, and NCI-H2452 cells. MATERIALS AND METHOD: For the adenovirus, the first mRNA/protein to be made (~1 h after infection) is E1A. Ad5F35 and Ad5 CRAD vectors containing the E1 gene controlled by the human midkine promoter (Ad5F35/MKp-E1 and Ad5/MKp-E1, respectively) were constructed. Western blotting and cell viability assays were carried out in cells transfected with Ad5/MKp-E1 and Ad5F35/MKp-E1. RESULTS: Coxsackie and adenovirus receptor (CAR), a cell surface target of Ad5, and CD46, a cell surface target of Ad35, were expressed in all the malignant mesothelioma cell lines examined here, as much as in HEK293 cells, with no significant differences in the expression levels among cells. Both Ad5/MKp-E1 and Ad5F35/MKp-E1 induced oncolysis of malignant mesothelioma cells in a viral particle-dependent manner, with similar efficacy. CONCLUSION: The results of the present study suggest that both Ad5/MKp-E1 and Ad5F35/MKp-E1 are useful for the gene therapy of human malignant mesothelioma.


Assuntos
Adenovírus Humanos/fisiologia , Terapia Genética/métodos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/terapia , Mesotelioma/genética , Mesotelioma/terapia , Terapia Viral Oncolítica/métodos , Proteínas E1 de Adenovirus/genética , Adenovírus Humanos/genética , Linhagem Celular Tumoral , Proteína de Membrana Semelhante a Receptor de Coxsackie e Adenovirus/biossíntese , Células HEK293 , Humanos , Neoplasias Pulmonares/virologia , Proteína Cofatora de Membrana/biossíntese , Mesotelioma/virologia , Midkina , Fatores de Crescimento Neural/genética , Regiões Promotoras Genéticas , Transfecção , Replicação Viral
10.
Tumori ; 98(2): 210-4, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22677986

RESUMO

AIMS AND BACKGROUND: Malignant mesothelioma is a fatal cancer of increasing incidence in north-eastern Italy. Together with asbestos, the polyomavirus SV40 was hypothesized to contribute to the onset of malignant mesothelioma. To investigate the putative role of SV40 in the individual susceptibility to asbestos-induced malignant mesothelioma, we conducted a molecular epidemiological study on a series of malignant mesothelioma patients from an area in north-eastern Italy hyperendemic for malignant pleural mesothelioma. METHODS AND STUDY DESIGN: We collected 63 mesothelioma samples from incidence cases of patients diagnosed with malignant pleural mesothelioma in the period 2009-2010. DNA was extracted from patients' tissue biopsies using the BioRobot EZ1 Qiagen workstation. SV40 sequence detection and quantification was performed by specific real time PCR. The 74.6% of the 63 enrolled patients had a history of asbestos exposure. The epithelioid histotype was more prevalent in males (64.0%) and the mixed in females (61.5%) who showed significantly higher cancer co-morbidity (46.1% vs 12%, P = 0.005). SV40 was detected in 22% of MM tumors, with a low viral load. In SV40-positive patients, a threefold increased risk of asbestos exposure was observed, more evident in females (OR 4.32) than in males (OR 1.20). CONCLUSIONS: Our findings indicate that a high prevalence of SV40 was present in malignant mesothelioma incident cases from an area hyperendemic for malignant mesothelioma in north-eastern Italy. Although asbestos is considered the main risk factor in malignant mesothelioma onset, a role for SV40 could be hypothesized.


Assuntos
Asbestos/toxicidade , Carcinógenos/toxicidade , Mesotelioma/epidemiologia , Mesotelioma/etiologia , Neoplasias Pleurais/epidemiologia , Neoplasias Pleurais/etiologia , Infecções por Polyomavirus/complicações , Vírus 40 dos Símios , Infecções Tumorais por Vírus/complicações , Adulto , Idoso , DNA Viral/isolamento & purificação , Suscetibilidade a Doenças , Doenças Endêmicas , Feminino , Humanos , Itália/epidemiologia , Masculino , Mesotelioma/induzido quimicamente , Mesotelioma/virologia , Pessoa de Meia-Idade , Neoplasias Pleurais/induzido quimicamente , Neoplasias Pleurais/virologia , Infecções por Polyomavirus/virologia , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Risco , Vírus 40 dos Símios/isolamento & purificação , Infecções Tumorais por Vírus/virologia , Carga Viral
11.
Expert Rev Respir Med ; 5(5): 683-97, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21955238

RESUMO

Simian virus 40 (SV40) is a DNA virus isolated in 1960 from contaminated polio vaccines, that induces mesotheliomas, lymphomas, brain and bone tumors, and sarcomas, including osteosarcomas, in hamsters. These same tumor types have been found to contain SV40 DNA and proteins in humans. Mesotheliomas and brain tumors are the two tumor types that have been most consistently associated with SV40, and the range of positivity has varied about from 6 to 60%, although a few reported 100% of positivity and a few reported 0%. It appears unlikely that SV40 infection alone is sufficient to cause human malignancy, as we did not observe an epidemic of cancers following the administration of SV40-contaminated vaccines. However, it seems possible that SV40 may act as a cofactor in the pathogenesis of some tumors. In vitro and animal experiments showing cocarcinogenicity between SV40 and asbestos support this hypothesis.


Assuntos
Neoplasias Encefálicas/virologia , Transformação Celular Viral , Mesotelioma/virologia , Infecções por Polyomavirus/virologia , Vírus 40 dos Símios/patogenicidade , Infecções Tumorais por Vírus/virologia , Animais , DNA Viral/isolamento & purificação , Humanos , Infecções por Polyomavirus/complicações , Medição de Risco , Fatores de Risco , Vírus 40 dos Símios/genética , Infecções Tumorais por Vírus/complicações
12.
Cancer Gene Ther ; 18(8): 571-8, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21660062

RESUMO

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.


Assuntos
Terapia Genética/métodos , Mesotelioma/terapia , Mesotelioma/virologia , Terapia Viral Oncolítica/métodos , Retroviridae/genética , Adulto , Animais , Linhagem Celular Tumoral , Citosina Desaminase/genética , Citosina Desaminase/metabolismo , Modelos Animais de Doenças , Feminino , Flucitosina/administração & dosagem , Flucitosina/farmacocinética , Humanos , Masculino , Mesotelioma/genética , Mesotelioma/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Pró-Fármacos/administração & dosagem , Pró-Fármacos/farmacocinética , Retroviridae/fisiologia , Análise de Sobrevida , Transdução Genética , Replicação Viral
13.
Mol Ther ; 19(1): 140-9, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20978474

RESUMO

Despite having potent oncolytic activity, in vitro, direct intratumoral injection of oncolytic vesicular stomatitis virus (VSV) into established AE17ova mesothelioma tumors in C57Bl/6 mice had no therapeutic effect. During studies to combine systemic cyclophosphamide (CPA) with VSV to suppress the innate immune reaction against VSV, we observed that CPA alone had highly significant antitumor effects in this model. However, against our expectations, the combination of CPA and VSV consistently reduced therapeutic efficacy compared to CPA alone, despite the fact that the combination increased intratumoral VSV titers. We show here that CPA-mediated therapy against AE17ova tumors was immune-mediated and dependent upon both CD4 T cells and natural killer (NK) cells. However, intratumoral VSV induced a transforming growth factor-ß (TGF-ß)-dependent suppressive activity, mediated by CD11b(+)GR-1(+) cells that significantly inhibited both antigen-specific T-cell activation, and CPA-activated, NK-dependent killing of AE17ova tumor cells. Overall, our results show that treatment with oncolytic viruses can induce a variety of immune-mediated consequences in vivo with both positive, or negative, effects on antitumor therapy. These underexplored immune consequences of treatment with oncolytic viruses may have significant, and possibly unexpected, impacts on how virotherapy interacts in combination with other agents which modulate antitumor immune effectors.


Assuntos
Ciclofosfamida/farmacologia , Terapia Genética/métodos , Mesotelioma/imunologia , Mesotelioma/terapia , Terapia Viral Oncolítica/métodos , Vírus Oncolíticos/fisiologia , Vírus da Estomatite Vesicular Indiana/fisiologia , Animais , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/virologia , Terapia Combinada , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/virologia , Melanoma Experimental/imunologia , Melanoma Experimental/terapia , Melanoma Experimental/virologia , Mesotelioma/tratamento farmacológico , Mesotelioma/virologia , Camundongos , Camundongos Endogâmicos C57BL , Vírus Oncolíticos/genética , Vírus Oncolíticos/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Vírus da Estomatite Vesicular Indiana/genética , Vírus da Estomatite Vesicular Indiana/imunologia , Vírus da Estomatite Vesicular Indiana/metabolismo , Replicação Viral
14.
Cancer Gene Ther ; 17(8): 550-8, 2010 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-20379224

RESUMO

Mesothelioma usually leads to death within 8-14 months of diagnosis. To increase the potency of oncolytic measles viruses (MVs) for mesothelioma therapy, we inserted the interferon beta (IFNbeta) gene alone or with the human thyroidal sodium iodide symporter (NIS) gene into attenuated MV of the Edmonston lineage. The corresponding mouse IFNbeta (mIFNbeta) viruses, MV-mIFNbeta and MV-mIFNbeta-NIS, successfully propagated in human mesothelioma cells, leading to intercellular fusion and cell death. High levels of mIFNbeta were detected in the supernatants of the infected cells, and radioiodine uptake was substantial in the cells infected with MV-mIFNbeta-NIS. MV with mIFNbeta expression triggered CD68-positive immune cell infiltration 2-4 times higher than MV-GFP injected into the tumor site. The numbers of CD31-positive vascular endothelial cells within the tumor were decreased at day 7 after intratumoral injection of MV-mIFNbeta or MV-mIFNbeta-NIS, but not after MV-GFP and PBS administration. Immunohistochemical analysis showed that MV-mIFNbeta changed the microenvironment of the mesothelioma by increasing innate immune cell infiltration and inhibiting tumor angiogenesis. Oncolytic MVs coding for IFNbeta effectively retarded growth of human mesotheliomas and prolonged survival time in several mesothelioma tumor models. The results suggest that oncolytic MVs that code for IFNbeta and NIS will be potent and versatile agents for the treatment of human mesothelioma.


Assuntos
Interferon beta/genética , Vírus do Sarampo/fisiologia , Mesotelioma/terapia , Terapia Viral Oncolítica/métodos , Simportadores/genética , Animais , Linhagem Celular Tumoral , Chlorocebus aethiops , Modelos Animais de Doenças , Feminino , Humanos , Interferon beta/biossíntese , Radioisótopos do Iodo , Vírus do Sarampo/genética , Mesotelioma/diagnóstico por imagem , Mesotelioma/virologia , Camundongos , Camundongos Nus , Camundongos SCID , Plasmídeos/genética , Simportadores/biossíntese , Tomografia Computadorizada de Emissão de Fóton Único/métodos , Células Vero , Ensaios Antitumorais Modelo de Xenoenxerto
15.
Tumori ; 96(5): 667-73, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-21302609

RESUMO

BACKGROUND AND AIM: Malignant mesothelioma is predominantly caused by asbestos exposure, although the association of Simian virus 40 in its pathogenesis is currently still under debate. Simian virus 40, a DNA rhesus monkey virus with oncogenic properties, accidentally contaminated early batches of polio vaccine in the 1960s. In the 1990s, viral sequences and proteins were discovered in several human tumors, which triggered research to find a link between Simian virus 40 and human cancers, especially malignant mesothelioma. The aim of our study was to establish an effective laboratory procedure for Simian virus 40 detection and to investigate the presence of Simian virus 40 DNA and small t antigen in mesothelioma samples from Slovenian patients. METHODS AND STUDY DESIGN: Paraffin-embedded malignant pleural mesothelioma specimens from 103 Slovenian patients were collected and used for total DNA isolation and real-time polymerase chain reaction for Simian virus 40 small t and large T DNA analysis. Special attention was devoted to primer design, good laboratory practice and polymerase chain reaction contamination prevention. Polymerase chain reaction products were sequenced and BLAST aligned. One 5 microm thick paraffin section from each patient's tissue block was stained with hematoxylin and eosin for histological typing and one for immunohistochemical detection of Simian virus 40 small t antigen using a monoclonal antibody against Simian virus 40 (Pab280). SV40-expressing Wi-38 cells were used as positive control in both PCR and immunohistochemistry. RESULTS: In real-time polymerase chain reaction analyses, only 4 samples gave products with primer pairs amplifying small t antigen and were inconsistent and poorly reproducible. BLAST alignment showed no homology with any deposited SV40 sequences. No immunopositive staining for SV40 small t antigen was found in any of the samples. CONCLUSIONS: We found no evidence of SV40 presence in tissue samples from 103 Slovenian patients with malignant pleural mesothelioma. Asbestos exposure remains the main risk factor for malignant pleural mesothelioma in Slovenia.


Assuntos
Antígenos Virais de Tumores/análise , Mesotelioma/epidemiologia , Neoplasias Pleurais/epidemiologia , Vírus 40 dos Símios/isolamento & purificação , DNA Viral/isolamento & purificação , Humanos , Imuno-Histoquímica , Mesotelioma/virologia , Neoplasias Pleurais/virologia , Reação em Cadeia da Polimerase , Vírus 40 dos Símios/genética , Vírus 40 dos Símios/imunologia , Eslovênia/epidemiologia
16.
Hum Gene Ther ; 21(1): 51-64, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19715403

RESUMO

Abstract Vesicular stomatitis virus (VSV) has shown promise as an oncolytic agent, although unmodified VSV can be neurotoxic. To avoid toxicity, a vector was created by introducing the interferon-beta (IFN-beta) gene (VSV.IFN-beta). We conducted this study to determine the ability of VSV.IFN-beta to lyse human cancer (mesothelioma) cells and to evaluate the potential of this recombinant virus for clinical translation. Four normal human mesothelial and 12 mesothelioma cell lines were tested for their susceptibility to VSV vectors in vitro. VSV.hIFN-beta did not cause cytotoxicity in any normal lines. Only 4 of 12 lines were effectively lysed by VSV.hIFN-beta. In the eight resistant lines, pretreatment with IFN-beta prevented lysis of cells by VSV.GFP, and VSV infection or addition of IFN-beta protein resulted in the upregulation of double-stranded RNA-dependent protein kinase (PKR), myxovirus resistance A (MxA), and 2',5'-oligo-adenylate-synthetase (2'5'-OAS) mRNA. In the susceptible lines, there was no protection by pretreatment with IFN-beta protein and no IFN- or VSV-induced changes in PKR, MxA, and 2'5'-OAS mRNA. This complete lack of IFN responsiveness could be explained by marked downregulation of interferon alpha receptors (IFNARs), p48, and PKR in both the mesothelioma cell lines and primary tumor biopsies screened. Presence of p48 in three tumor samples predicted responsiveness to IFN. Our data indicate that many mesothelioma tumors have partially intact IFN pathways that may affect the efficacy of oncolytic virotherapy. However, it may be feasible to prescreen individual susceptibility to VSV.IFN-beta by immunostaining for the presence of p48 protein.


Assuntos
Vetores Genéticos/genética , Interferon beta/uso terapêutico , Mesotelioma/genética , Mesotelioma/terapia , Terapia Viral Oncolítica/métodos , Neoplasias Pleurais/genética , Neoplasias Pleurais/terapia , Vesiculovirus/genética , Animais , Linhagem Celular Tumoral , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Heterogeneidade Genética/efeitos dos fármacos , Proteínas de Fluorescência Verde/metabolismo , Humanos , Interferon beta/genética , Interferon beta/farmacologia , Mesotelioma/patologia , Mesotelioma/virologia , Camundongos , Camundongos SCID , Proteínas de Neoplasias/metabolismo , Neoplasias Pleurais/patologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transdução de Sinais/efeitos dos fármacos , Resultado do Tratamento , Vesiculovirus/fisiologia , Carga Viral , Replicação Viral/efeitos dos fármacos
17.
J Clin Virol ; 47(2): 196-8, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-20006539

RESUMO

BACKGROUND: Merkel cell polyomavirus (MCPyV) is the first polyoma virus consistently linked to the etiology of a human cancer. Serological studies indicate that the virus is commonly acquired in childhood, with seroprevalence reaching 50% or higher among young adults. The modes of MCPyV transmission are still unclear, but it has been identified in respiratory tract samples. Given its respiratory tropism, we examined whether MCPyV could be detected in mesothelioma tissue, a malignancy induced in animal models by another polyomavirus, SV40. OBJECTIVE: To determine if MCPyV DNA can be detected in mesothelioma. STUDY DESIGN: DNA was extracted from 45 fresh-frozen mesothelioma samples. PCR was used to detect and quantify the abundance of MCPyV DNA, and a human control gene, in duplicates of the tissues. DNA from a sequence verified MCC tumor was used as a positive control. RESULTS: The human control gene was detected at high levels in all but three mesothelioma tissues. MCPyV DNA was detected in only one mesothelioma, and the level of viral DNA was very low. CONCLUSIONS: These results are inconsistent with the hypothesis that MCPyV is etiologically linked to mesothelioma.


Assuntos
DNA Viral/isolamento & purificação , Mesotelioma/etiologia , Mesotelioma/virologia , Polyomavirus/isolamento & purificação , Adolescente , Criança , Pré-Escolar , Humanos , Adulto Jovem
18.
Cancer Res ; 69(19): 7713-20, 2009 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-19773437

RESUMO

Our preclinical and clinical trials using a replication-defective adenoviral vector expressing IFN-beta have shown promising results for the treatment of malignant mesothelioma. Based on the hypotheses that a replication-competent vesicular stomatitis virus (VSV) oncolytic vector would transduce more tumor cells in vivo, that coexpression of the immunostimulatory IFN-beta gene would enhance the immune-based effector mechanisms associated both with regression of mesotheliomas and with VSV-mediated virotherapy, and that virus-derived IFN-beta would add further safety to the VSV platform, we tested the use of IFN-beta as a therapeutic transgene expressed from VSV as a novel treatment for mesothelioma. VSV-IFN-beta showed significant therapy against AB12 murine mesotheliomas in the context of both local and locoregional viral delivery. Biologically active IFN-beta expressed from VSV added significantly to therapy compared with VSV alone, dependent in part on host CD8+ T-cell responses. Immune monitoring suggested that these antitumor T-cell responses may be due to a generalized T-cell activation rather than the priming of tumor antigen-specific T-cell responses. Finally, IFN-beta also added considerable extra safety to the virus by providing protection from off-target viral replication in nontumor tissues and protected severe combined immunodeficient mice from developing lethal neurotoxicity. The enhanced therapeutic index provided by the addition of IFN-beta to VSV therefore provides a powerful justification for the development of this virus for future clinical trials.


Assuntos
Interferon beta/genética , Mesotelioma/terapia , Terapia Viral Oncolítica/métodos , Vesiculovirus/genética , Animais , Linhagem Celular Tumoral , Epitopos de Linfócito T/imunologia , Terapia Genética/métodos , Humanos , Interferon beta/biossíntese , Interferon beta/imunologia , Mesotelioma/genética , Mesotelioma/imunologia , Mesotelioma/virologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos SCID , Doenças do Sistema Nervoso/virologia , Linfócitos T/imunologia
19.
Clin Cancer Res ; 15(11): 3791-801, 2009 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-19470726

RESUMO

PURPOSE: Oncolytic viral therapy continues to be investigated for the treatment of cancer, and future studies in patients would benefit greatly from a noninvasive modality for assessing virus dissemination, targeting, and persistence. The purpose of this study was to determine if a genetically modified vaccinia virus, GLV-1h99, containing a human norepinephrine transporter (hNET) reporter gene, could be sequentially monitored by [(123)I]metaiodobenzylguanidine (MIBG) gamma-camera and [(124)I]MIBG positron emission tomography (PET) imaging. EXPERIMENTAL DESIGN: GLV-1h99 was tested in human malignant mesothelioma and pancreatic cancer cell lines for cytotoxicity, expression of the hNET protein using immunoblot analysis, and [(123)I]MIBG uptake in cell culture assays. In vivo [(123)I]MIBG gamma-camera and serial [(124)I]MIBG PET imaging was done in MSTO-211H orthotopic pleural mesothelioma tumors. RESULTS: GLV-1h99 successfully infected and provided dose-dependent levels of transgene hNET expression in human malignant mesothelioma and pancreatic cancer cells. The time course of [(123)I]MIBG accumulation showed a peak of radiotracer uptake at 48 hours after virus infection in vitro. In vivo hNET expression in MSTO-211H pleural tumors could be imaged by [(123)I]MIBG scintigraphy and [(124)I]MIBG PET 48 and 72 hours after GLV-1h99 virus administration. Histologic analysis confirmed the presence of GLV-1h99 in tumors. CONCLUSION: GLV-1h99 shows high mesothelioma tumor cell infectivity and cytotoxic efficacy. The feasibility of imaging virus-targeted tumor using the hNET reporter system with [(123)I]MIBG gamma-camera and [(124)I]MIBG PET was shown in an orthotopic pleural mesothelioma tumor model. The inclusion of human reporter genes into recombinant oncolytic viruses enhances the potential for translation to clinical monitoring of oncolytic viral therapy.


Assuntos
Neoplasias Experimentais/metabolismo , Proteínas da Membrana Plasmática de Transporte de Norepinefrina/metabolismo , Vírus Oncolíticos/metabolismo , Vírus Vaccinia/metabolismo , 3-Iodobenzilguanidina/farmacocinética , Animais , Linhagem Celular Tumoral , Sobrevivência Celular , Câmaras gama , Engenharia Genética , Humanos , Immunoblotting , Radioisótopos do Iodo , Masculino , Mesotelioma/metabolismo , Mesotelioma/patologia , Mesotelioma/virologia , Camundongos , Camundongos Nus , Neoplasias Experimentais/diagnóstico por imagem , Neoplasias Experimentais/patologia , Proteínas da Membrana Plasmática de Transporte de Norepinefrina/genética , Vírus Oncolíticos/genética , Vírus Oncolíticos/fisiologia , Neoplasias Pancreáticas/metabolismo , Neoplasias Pancreáticas/patologia , Neoplasias Pancreáticas/virologia , Tomografia por Emissão de Pósitrons , Transplante Heterólogo , Vírus Vaccinia/genética , Vírus Vaccinia/fisiologia
20.
Am J Pathol ; 174(6): 2324-36, 2009 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-19435792

RESUMO

The calcium-binding protein calretinin has emerged as a useful marker for the identification of mesotheliomas of the epithelioid and mixed types, but its putative role in tumor development has not been addressed previously. Although exposure to asbestos fibers is considered the main cause of mesothelioma, undoubtedly, not all mesothelioma patients have a history of asbestos exposure. The question as to whether the SV40 virus is involved as a possible co-factor is still highly debated. Here we show that increased expression of SV40 early gene products in the mesothelial cell line MeT-5A induces the expression of calretinin and that elevated calretinin levels strongly correlate with increased resistance to asbestos cytotoxicity. Calretinin alone mediates a significant part of this protective effect because cells stably transfected with calretinin cDNA were clearly more resistant to the toxic effects of crocidolite than mock-transfected control cells. Down-regulation of calretinin by antisense methods restored the sensitivity to asbestos toxicity to a large degree. The protective effect observed in clones with higher calretinin expression levels could be eliminated by phosphatidylinositol 3-kinase (PI3K) inhibitors, implying an important role for the PI3K/AKT signaling (survival) pathway in mediating the protective effect. Up-regulation of calretinin, resulting from either asbestos exposure or SV40 oncoproteins, may be a common denominator that leads to increased resistance to asbestos cytotoxicity and thereby contributes to mesothelioma carcinogenesis.


Assuntos
Asbesto Crocidolita/efeitos adversos , Transformação Celular Neoplásica/metabolismo , Mesotelioma/induzido quimicamente , Mesotelioma/virologia , Infecções por Polyomavirus/metabolismo , Proteína G de Ligação ao Cálcio S100/metabolismo , Antígenos Transformantes de Poliomavirus , Western Blotting , Calbindina 2 , Linhagem Celular Tumoral , Transformação Celular Neoplásica/induzido quimicamente , Expressão Gênica , Humanos , Imuno-Histoquímica , Fosfatidilinositol 3-Quinases/metabolismo , Infecções por Polyomavirus/complicações , Proteínas Proto-Oncogênicas c-akt/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais/fisiologia , Vírus 40 dos Símios , Transfecção , Infecções Tumorais por Vírus/complicações , Infecções Tumorais por Vírus/metabolismo , Regulação para Cima
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