RESUMO
Raltegravir (RAL), a human immunodeficiency virus (HIV)-1 integrase inhibitor, has been administered as part of antiretroviral therapy. Studies in patients with HIV-1 have shown high variability in the pharmacokinetics of RAL, and in healthy volunteers, coadministration of proton-pump inhibitors has been shown to increase the plasma RAL concentrations. Here, we found that RAL containing a 1,3,4-oxadiazole ring is converted to a hydrolysis product (H-RAL) with a cleaved 1,3,4-oxadiazole ring at pH 1.0 and 13.0 conditions in vitro, thereby reducing the anti-HIV activity of the drug. The inclusion of cyclodextrins (beta-cyclodextrin [ßCD], random methyl-ßCD [RAM-ßCD], and hydroxypropyl-ßCD [HP-ßCD]) can protect RAL from pH-induced changes. The conversion of RAL to H-RAL was detected by using various mass spectrometry analyses. The chromatogram of H-RAL increased in a time-dependent manner similar to another 1,3,4-oxadiazole-containing drug, zibotentan, using high-performance liquid chromatography. Oral bioavailability and target protein interactions of H-RAL were predicted to be lower than those of RAL. Moreover, H-RAL exhibited significantly reduced anti-HIV-1 activity, whereas combinations with ßCD, RAM-ßCD, and HP-ßCD attenuated this effect in cell-based assays. These findings suggest that ßCDs can potentially protect against the conversion of RAL to H-RAL under acidic conditions in the stomach, thereby preserving the anti-HIV-1 effect of RAL. Although clinical trials are needed for evaluation, we anticipate that protective devices such as ßCDs may improve the pharmacokinetics of RAL, leading to better treatment outcomes, including reduced dosing, long-term anti-HIV-1 activity, and deeper HIV-1 suppression.
RESUMO
BACKGROUND: Human T-cell leukemia virus type 1 (HTLV-1) causes chronic infection leading to development of adult T-cell leukemia (ATL) and inflammatory diseases. Non-human primates infected with simian T-cell leukemia virus type 1 (STLV-1) are considered to constitute a suitable animal model for HTLV-1 research. However, the function of the regulatory and accessory genes of STLV-1 has not been analyzed in detail. In this study, STLV-1 in naturally infected Japanese macaques was analyzed. RESULTS: We identified spliced transcripts of STLV-1 corresponding to HTLV-1 tax and HTLV-1 bZIP factor (HBZ). STLV-1 Tax activated the NFAT, AP-1 and NF-κB signaling pathways, whereas STLV-1 bZIP factor (SBZ) suppressed them. Conversely, SBZ enhanced TGF-ß signaling and induced Foxp3 expression. Furthermore, STLV-1 Tax activated the canonical Wnt pathway while SBZ suppressed it. STLV-1 Tax enhanced the viral promoter activity while SBZ suppressed its activation. Then we addressed the clonal proliferation of STLV-1⺠cells by massively sequencing the provirus integration sites. Some clones proliferated distinctively in monkeys with higher STLV-1 proviral loads. Notably, one of the monkeys surveyed in this study developed T-cell lymphoma in the brain; STLV-1 provirus was integrated in the lymphoma cell genome. When anti-CCR4 antibody, mogamulizumab, was administered into STLV-1-infected monkeys, the proviral load decreased dramatically within 2 weeks. We observed that some abundant clones recovered after discontinuation of mogamulizumab administration. CONCLUSIONS: STLV-1 Tax and SBZ have functions similar to those of their counterparts in HTLV-1. This study demonstrates that Japanese macaques naturally infected with STLV-1 resemble HTLV-1 carriers and are a suitable model for the investigation of persistent HTLV-1 infection and asymptomatic HTLV-1 carrier state. Using these animals, we verified that mogamulizumab, which is currently used as a drug for relapsed ATL, is also effective in reducing the proviral load in asymptomatic individuals.
Assuntos
Infecções por Deltaretrovirus/veterinária , Modelos Animais de Doenças , Leucemia de Células T/veterinária , Doenças dos Primatas/patologia , Doenças dos Primatas/virologia , Vírus Linfotrópico T Tipo 1 de Primatas/isolamento & purificação , Infecções Tumorais por Vírus/veterinária , Animais , Infecções por Deltaretrovirus/patologia , Infecções por Deltaretrovirus/virologia , Humanos , Leucemia de Células T/patologia , Leucemia de Células T/virologia , Macaca , Vírus Linfotrópico T Tipo 1 de Primatas/crescimento & desenvolvimento , Vírus Linfotrópico T Tipo 1 de Primatas/patogenicidade , Infecções Tumorais por Vírus/patologia , Infecções Tumorais por Vírus/virologiaRESUMO
Human T-cell leukemia virus type 1 (HTLV-1) belongs to Delta Retorviridae, and induces a malignancy of CD4+CD25+ T-cells, adult T-cell leukemia (ATL), and several chronic inflammatory diseases, such as HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP) and HTLV-1 uveitis. A nationwide survey of HTLV-1-infected subjects, which was recently conducted by Japanese government, revealed that the numbers of HTLV-1 carriers and patients with HTLV-1-associated diseases have not decreased much over the last two decades in Japan. In contrast, novel findings on HTLV-1 dynamics in vivo and molecular mechanisms of its pathogenesis are accumulating by detailed analysis of newly identified viral and cellular factors, novel technologies such as next-generation sequencing, and appropriate animal models for HTLV-1 research. In this review, we summarize the recent progress of HTLV-1 research.
Assuntos
Vírus Linfotrópico T Tipo 1 Humano , Animais , Fatores de Transcrição de Zíper de Leucina Básica/genética , Portador Sadio/epidemiologia , Genes pX/genética , Vírus Linfotrópico T Tipo 1 Humano/genética , Vírus Linfotrópico T Tipo 1 Humano/patogenicidade , Humanos , Imunoterapia , Japão/epidemiologia , Leucemia-Linfoma de Células T do Adulto/epidemiologia , Leucemia-Linfoma de Células T do Adulto/genética , Leucemia-Linfoma de Células T do Adulto/imunologia , Leucemia-Linfoma de Células T do Adulto/virologia , Análise de Sequência com Séries de Oligonucleotídeos , Paraparesia Espástica Tropical/epidemiologia , Paraparesia Espástica Tropical/virologia , RNA Viral , Proteínas dos Retroviridae , Uveíte/virologia , Proteínas Virais/genética , Proteínas Virais Reguladoras e Acessórias/genética , Proteínas Virais Reguladoras e Acessórias/fisiologiaRESUMO
BACKGROUND: Human T-cell Leukemia Virus type 1 (HTLV-1) infects 20 million individuals world-wide and causes Adult T-cell Leukemia/Lymphoma (ATLL), a highly aggressive T-cell cancer. ATLL is refractory to treatment with conventional chemotherapy and fewer than 10% of afflicted individuals survive more than 5 years after diagnosis. HTLV-1 encodes a viral oncoprotein, Tax, that functions in transforming virus-infected T-cells into leukemic cells. All ATLL cases are believed to have reduced p53 activity although only a minority of ATLLs have genetic mutations in their p53 gene. It has been suggested that p53 function is inactivated by the Tax protein. RESULTS: Using genetically altered mice, we report here that Tax expression does not achieve a functional equivalence of p53 inactivation as that seen with genetic mutation of p53 (i.e. a p53 -/- genotype). Thus, we find statistically significant differences in tumorigenesis between Tax+p53 +/+ versus Tax+p53 -/- mice. We also find a role contributed by the cellular Wip1 phosphatase protein in tumor formation in Tax transgenic mice. Notably, Tax+Wip1 -/- mice show statistically significant reduced prevalence of tumorigenesis compared to Tax+Wip1 +/+ counterparts. CONCLUSIONS: Our findings provide new insights into contributions by p53 and Wip1 in the in vivo oncogenesis of Tax-induced tumors in mice.
Assuntos
Transformação Celular Neoplásica/metabolismo , Transformação Celular Viral , Produtos do Gene tax/metabolismo , Fosfoproteínas Fosfatases/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Animais , Linhagem Celular Tumoral , Transformação Celular Neoplásica/genética , Transformação Celular Viral/genética , Regulação Neoplásica da Expressão Gênica , Regulação Viral da Expressão Gênica , Produtos do Gene tax/genética , Genótipo , Humanos , Camundongos , Camundongos Transgênicos , Neoplasias/genética , Neoplasias/metabolismo , Neoplasias/mortalidade , Fosfoproteínas Fosfatases/deficiência , Fosfoproteínas Fosfatases/genética , Fosforilação , Proteína Fosfatase 2C , Transcrição Gênica , Proteína Supressora de Tumor p53/genéticaRESUMO
NF-κB plays a key role in innate and acquired immunity. Its activity is regulated through intricate signaling networks. Persistent or excessive activation of NF-κB induces diseases, such as autoimmune disorders and malignant neoplasms. Infection by human T cell leukemia virus type 1 (HTLV-1) causes a fatal hematopoietic malignancy termed adult T cell leukemia (ATL). The HTLV-1 viral oncoprotein Tax functions pivotally in leukemogenesis through its potent activation of NF-κB. Recent findings suggest that protein ubiquitination is crucial for proper regulation of NF-κB signaling and for Tax activity. Here, we report that ubiquitin-specific peptidase USP20 deubiquitinates TRAF6 and Tax and suppresses interleukin 1ß (IL-1ß)- and Tax-induced NF-κB activation. Our results point to USP20 as a key negative regulator of Tax-induced NF-κB signaling.
Assuntos
Regulação da Expressão Gênica , Produtos do Gene tax/metabolismo , NF-kappa B/metabolismo , Transdução de Sinais , Fator 6 Associado a Receptor de TNF/metabolismo , Ubiquitina Tiolesterase/metabolismo , Linhagem Celular , Regulação Viral da Expressão Gênica , Produtos do Gene tax/genética , Células HeLa , Vírus Linfotrópico T Tipo 1 Humano/genética , Vírus Linfotrópico T Tipo 1 Humano/metabolismo , Vírus Linfotrópico T Tipo 1 Humano/fisiologia , Humanos , Interleucina-1/genética , Interleucina-1/metabolismo , Células Jurkat , NF-kappa B/genética , Linfócitos T , Fator 6 Associado a Receptor de TNF/genética , Ubiquitina Tiolesterase/genéticaRESUMO
Human T-cell leukemia virus type 1 (HTLV-1) is an oncogenic retrovirus etiologically causal of adult T-cell leukemia (ATL). The virus encodes a Tax oncoprotein that functions in transcriptional regulation, cell cycle control, and transformation. ATL is a highly virulent cancer that is resistant to chemotherapeutic treatments. To understand this disease better, it is important to comprehend how HTLV-1 promotes cellular growth and survival. Tax activation of NF-kappaB is important for the proliferation and transformation of virus-infected cells. We show here that prolyl isomerase Pin1 is over expressed in HTLV-1 cell lines; Pin1 binds Tax and regulates Tax-induced NF-kappaB activation.
Assuntos
Produtos do Gene tax/metabolismo , Vírus Linfotrópico T Tipo 1 Humano/fisiologia , NF-kappa B/metabolismo , Peptidilprolil Isomerase/metabolismo , Linhagem Celular , Humanos , Peptidilprolil Isomerase de Interação com NIMA , Ligação ProteicaRESUMO
The spindle assembly checkpoint (SAC) guards against chromosomal missegregation during mitosis. To investigate the role of SAC in tumor development, mice heterozygously knocked out for the mitotic arrest deficient (Mad) genes Mad1 and/or Mad2 were mated with p53(+/) (-) mice. Increased tumor frequencies were reproducibly observed in Mad2(+/) (-)p53(+/) (-) (88.2%) and Mad1(+/) (-)Mad2(+/) (-)p53(+/) (-) (95.0%) mice compared with p53(+/) (-) (66.7%) mice. Moreover, 53% of Mad2(+/) (-)p53(+/) (-) mice developed lymphomas compared with 11% of p53(+/) (-) mice. By examining chromosome content, increased loss in diploidy was seen in cells from Mad2(+/) (-)p53(+/) (-) versus p53(+/) (-) mice, correlating loss of SAC function, in a p53(+/) (-) context, with increased aneuploidy and tumorigenesis. The findings here provide evidence for a cooperative role of Mad1/Mad2 and p53 genes in preventing tumor development.
Assuntos
Proteínas de Ciclo Celular/genética , Neoplasias/genética , Proteínas Serina-Treonina Quinases/deficiência , Fuso Acromático/patologia , Proteína Supressora de Tumor p53/deficiência , Animais , Cruzamentos Genéticos , Feminino , Fase G1/genética , Frequência do Gene , Genótipo , Tamanho da Ninhada de Vivíparos , Proteínas Mad2 , Camundongos , Camundongos Knockout , Mutação de Sentido Incorreto , Neoplasias/patologia , Proteínas Serina-Treonina Quinases/genética , Fase S/genética , Proteína Supressora de Tumor p53/genéticaRESUMO
Human T-cell leukemia virus type I (HTLV-I) provirus load differs more than 100-fold among carriers and a high provirus load in the peripheral blood mononuclear cells (PBMCs) is regarded as a risk factor for both preleukemic states and inflammatory diseases including HTLV-I-associated myelopathy (HAM). We examined polymorphisms in the genes for tumor necrosis factor (TNF), TNF receptor type 1 and 2, lymphotoxin (LT)-alpha, interleukin (IL)-1beta, IL-6, IL-10, monocyte chemoattractant protein (MCP)-1, and mannose binding protein (ManBP) in 143 HTLV-I carriers whether these polymorphisms affect the provirus load in the PBMCs of carriers. No significant association was observed between these polymorphisms and the provirus load. Homozygotes for a ManBP-variant allele, however, showed a tendency for the decreased number of provirus load. When combined, the data on the alleles of LT-alpha and MCP-1, HTLV-I carriers having high producer alleles of both genes showed a trend for increased provirus load. These data suggest that inflammation or an active immune response may induce an increased amount of HTLV-I-infected T cells, leading to a high provirus load.
Assuntos
Citocinas/genética , Vírus Linfotrópico T Tipo 1 Humano/isolamento & purificação , Lectina de Ligação a Manose/genética , Polimorfismo Genético , Alelos , Povo Asiático , Portador Sadio , Rearranjo Gênico do Linfócito T , Infecções por HTLV-I/diagnóstico , Humanos , Provírus/isolamento & purificação , Carga ViralRESUMO
By focusing on the a-g interactions, successful design and selection were accomplished to obtain a leucine-zipper segment that discriminates the appropriate partner over another that provides very similar patterns of electrostatic interactions.
Assuntos
Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Peptídeos/metabolismo , Fatores de Transcrição de Zíper de Leucina Básica/química , Células HeLa , Humanos , Peptídeos/química , Eletricidade EstáticaRESUMO
Human T cell leukemia virus type 1 (HTLV-1) is an etiological pathogen of several human diseases, including adult T-cell leukemia (ATL), HTLV-1-associated myelopathy (HAM)/tropical spastic paraparesis (TSP), and inflammatory disorders such as uveitis and dermatitis. HTLV-1 spreads mainly through cell-to-cell transmission, induces clonal proliferation of infected T cells in vivo, and after a long latent period, a subset of HTLV-1 carriers develop ATL. Understanding the molecular mechanisms of infection and oncogenesis is important for the development of new strategies of prophylaxis and molecular-targeted therapies, since ATL has a poor prognosis, despite intensive chemotherapy. In this review, we will summarize recent progress in HTLV-1 research, and especially novel findings on viral transmission and leukemogenic mechanisms by two viral oncogenes, HBZ and tax.
Assuntos
Fatores de Transcrição de Zíper de Leucina Básica , Genes pX , Infecções por HTLV-I/genética , Infecções por HTLV-I/virologia , Vírus Linfotrópico T Tipo 1 Humano/genética , Vírus Linfotrópico T Tipo 1 Humano/patogenicidade , Proteínas Virais , Animais , Fatores de Transcrição Forkhead/metabolismo , Infecções por HTLV-I/transmissão , Vírus Linfotrópico T Tipo 1 Humano/fisiologia , Humanos , Ativação Linfocitária/genética , Camundongos , Terapia de Alvo Molecular , NF-kappa B/metabolismo , Proteínas dos Retroviridae , Linfócitos T/imunologia , Fator de Transcrição AP-1/metabolismo , Montagem de Vírus , Integração ViralRESUMO
Human tumor viruses are associated with a variety of human malignancies, and it is estimated that 15% of all human cancers have a viral etiology. An abnormality in chromosomal ploidy or aneuploidy is a hallmark of cancers. In normal cells, euploidy is governed by several factors including an intact spindle assembly checkpoint, accurate centrosome duplication, and proper cytokinesis. Viral oncoproteins are suggested to perturb the cellular machineries for chromosomal segregation creating aneuploidy which can lead to the malignant transformation of infected cells. Here, we review in brief some of the mechanisms used by viruses that can cause cellular aneuploidy.