共查询到20条相似文献,搜索用时 0 毫秒
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Briana Jill Williams Shilpa Bhatia Lisa K. Adams Susan Boling Jennifer L. Carroll Xiao-Lin Li Donna L. Rogers Nikolay Korokhov Imre Kovesdi Alexander V. Pereboev David T. Curiel J. Michael Mathis 《PloS one》2012,7(10)
Human prostate tumor vaccine and gene therapy trials using ex vivo methods to prime dendritic cells (DCs) with prostate specific membrane antigen (PSMA) have been somewhat successful, but to date the lengthy ex vivo manipulation of DCs has limited the widespread clinical utility of this approach. Our goal was to improve upon cancer vaccination with tumor antigens by delivering PSMA via a CD40-targeted adenovirus vector directly to DCs as an efficient means for activation and antigen presentation to T-cells. To test this approach, we developed a mouse model of prostate cancer by generating clonal derivatives of the mouse RM-1 prostate cancer cell line expressing human PSMA (RM-1-PSMA cells). To maximize antigen presentation in target cells, both MHC class I and TAP protein expression was induced in RM-1 cells by transduction with an Ad vector expressing interferon-gamma (Ad5-IFNγ). Administering DCs infected ex vivo with CD40-targeted Ad5-huPSMA, as well as direct intraperitoneal injection of the vector, resulted in high levels of tumor-specific CTL responses against RM-1-PSMA cells pretreated with Ad5-IFNγ as target cells. CD40 targeting significantly improved the therapeutic antitumor efficacy of Ad5-huPSMA encoding PSMA when combined with Ad5-IFNγ in the RM-1-PSMA model. These results suggest that a CD-targeted adenovirus delivering PSMA may be effective clinically for prostate cancer immunotherapy. 相似文献
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Yao XL Yoshioka Y Ruan GX Chen YZ Mizuguchi H Mukai Y Okada N Gao JQ Nakagawa S 《Biomacromolecules》2012,13(8):2402-2409
We have previously developed a novel adenovirus vector (Adv) that targeted tumor tissues/vasculatures after systemic administration. The surface of this Adv is conjugated with CGKRK tumor homing peptide by the cross-linking reaction of polyethyleneglycol (PEG). In this study, we showed that the condition of PEG modification was important to minimize the gene expression in normal tissues after systemic treatment. When Adv was modified only with PEG-linked CGKRK, its luciferase expression was enhanced even in the liver tissue, as well as the tumor tissue. However, in the reaction with the mixture of non-cross-linking PEG and PEG-linked CGKRK, we found out that the best modification could suppress its gene expression in the liver, without losing that in the tumor. We also studied the internalization mechanisms of CGKRK-conjugated Adv. Results suggested that there is a specific interaction of the CGKRK peptide with a receptor at the cell surface enabling efficient internalization of CGKRK-conjugated Adv. The presence of cell-surface heparan sulfate is important receptor for the cellular binding and uptake of CGKRK-conjugated Adv. Moreover, macropinocytosis-mediated endocytosis is also important in endocytosis of CGKRK-conjugated Adv, aside from clathrin-mediated and caveolae-mediated endocytosis. These results could help evaluate the potentiality of CGKRK-conjugated Adv as a prototype vector with suitable efficacy and safety for systemic cancer gene therapy. 相似文献
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A Helper-Independent Adenovirus Vector with E1, E3, and Fiber Deleted: Structure and Infectivity of Fiberless Particles 总被引:3,自引:2,他引:3
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Dan J. Von Seggern Charles Y. Chiu Shonna Kaye Fleck Phoebe L. Stewart Glen R. Nemerow 《Journal of virology》1999,73(2):1601-1608
The adenovirus (Ad) fiber protein largely determines viral tropism through interaction with specific cell surface receptors. This molecule may also be involved in virion assembly or maturation, as some previously characterized fiber mutants were defective for processing of viral structural proteins. We previously described packaging cell lines that express Ad type 5 (Ad5) fiber and can complement the temperature-sensitive Ad fiber mutant H5ts142. We have now used these packaging cells to construct a new adenoviral vector (Ad5.βgal.ΔF) with E1, E3, and L5 (fiber) deleted and analyzed the fiber null phenotype. Ad5.βgal.ΔF growth was completely helper independent, and fiberless particles were produced by a single final round of growth in 293 cells. Cryoelectron microscopic studies and sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed that the structure and composition of these particles was nearly identical to those of first-generation Ad vectors. As expected, fiberless particles had reduced infectivity on epithelial cells, but they retained the ability to infect monocytic cells via an integrin-dependent pathway. These studies provide a novel approach to developing retargeted Ad gene therapy vectors. 相似文献
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Pancreatic adenocarcinoma or pancreatic cancer is often diagnosed at a very late stage at which point treatment options are minimal. Current chemotherapeutic interventions prolong survival marginally, thereby emphasizing the acute need for better treatment options to effectively manage this disease. Studies from different laboratories have shown that the Alzheimer disease-associated amyloid precursor protein (APP) is overexpressed in various cancers but its significance is not known. Here we sought to determine the role of APP in pancreatic cancer cell survival and proliferation. Our results show that pancreatic cancer cells secrete high levels of sAPPα, the α-secretase cleaved ectodomain fragment of APP, as compared with normal non-cancerous cells. Treatment of cells with batimastat or GI254023X, inhibitors of the α-secretase ADAM10, prevented sAPPα generation and reduced cell survival. Additionally, inhibition of sAPPα significantly reduced anchorage independent growth of the cancer cells. The effect of batimastat on cell survival and colony formation was enhanced when sAPPα downregulation was combined with gemcitabine treatment. Moreover, treatment of batimastat-treated cells with recombinant sAPPα reversed the inhibitory effect of the drug thereby indicating that sAPPα can indeed induce proliferation of cancer cells. Down-regulation of APP and ADAM10 brought about similar results, as did batimastat treatment, thereby confirming that APP processing is important for growth and proliferation of these cells. These results suggest that inhibition of sAPPα generation might enhance the effectiveness of the existing chemotherapeutic regimen for a better outcome. 相似文献
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目的:构建利用RNA干扰技术沉默HPV16E7基因的腺病毒载体,并探讨其对人宫颈癌细胞系CaSki细胞增殖的影响,以及腺病毒载体在宫颈癌基因治疗中的可行性。方法:利用腺病毒载体介导的RNA干扰对CaSki细胞中HPV16E7蛋白的表达进行抑制。显微镜观察细胞病变情况。MTT实验用来检测腺病毒载体感染的CaSki细胞的增殖情况。结果:成功构建了用以介导CaSki细胞中HPV16E7基因沉默的腺病毒载体。腺病毒感染后的CaSki细胞发生典型病理变化,HPV16E7基因表达受到明显抑制,细胞分裂明显减慢。结论:腺病毒载体介导的RNA干扰能够特异性的沉默HPV16E7基因,抑制CaSki细胞的增殖,为腺病毒载体用于宫颈癌基因治疗的可行性提供了依据。 相似文献
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A safe and efficacious cancer medicine is necessary due to the increasing population of cancer patients whose particular diseases cannot be cured by the currently available treatment. Adenoviral (Ad) vectors represent a promising therapeutic medicine for human cancer therapy. However, several improvements are needed in order for Ad vectors to be effective cancer therapeutics, which include, but are not limited to, improvement of cellular uptake, enhanced cancer cell killing activity, and the capability of vector visualization and tracking once injected into the patients. To this end, we attempted to develop an Ad as a multifunctional platform incorporating targeting, imaging, and therapeutic motifs. In this study, we explored the utility of this proposed platform by generating an Ad vector containing the poly-lysine (pK), the herpes simplex virus type 1 (HSV-1) thymidine kinase (TK), and the monomeric red fluorescent protein (mRFP1) as targeting, tumor cell killing, and imaging motifs, respectively. Our study herein demonstrates the generation of the triple mosaic Ad vector with pK, HSV-1 TK, and mRFP1 at the carboxyl termini of Ad minor capsid protein IX (pIX). In addition, the functionalities of pK, HSV-1 TK, and mRFP1 proteins on the Ad vector were retained as confirmed by corresponding functional assays, indicating the potential multifunctional application of this new Ad vector for cancer gene therapy. The validation of the triple mosaic Ad vectors also argues for the ability of pIX modification as a base for the development of multifunctional Ad vectors. 相似文献
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Jinsheng Hong Zhenhuan Zhang Wenlong Lv Mei Zhang Chun Chen Shanmin Yang Shan Li Lurong Zhang Deping Han Weijian Zhang 《PloS one》2013,8(8)
Icaritin (ICT) is a hydrolytic form of icariin isolated from plants of the genus Epimedium. This study was to investigate the radiosensitization effect of icaritin and its possible underlying mechanism using murine 4T1 breast cancer cells. The combination of Icaritin at 3 µM or 6 µM with 6 or 8 Gy of ionizing radiation (IR) in the clonogenic assay yielded an ER (enhancement ratio) of 1.18 or 1.28, CI (combination index) of 0.38 or 0.19 and DRI (dose reducing index) of 2.51 or 5.07, respectively. These strongly suggest that Icaritin exerted a synergistic killing (?) effect with radiation on the tumor cells. This effect might relate with bioactivities of ICT: 1) exert an anti-proliferative effect in a dose- and time-dependent manner, which is different from IR killing effect but likely work together with the IR effect; 2) suppress the IR-induced activation of two survival paths, ERK1/2 and AKT; 3) induce the G2/M blockage, enhancing IR killing effect; and 4) synergize with IR to enhance cell apoptosis. In addition, ICT suppressed angiogenesis in chick embryo chorioallantoic membrane (CAM) assay. Taken together, ICT is a new radiosensitizer and can enhance anti-cancer effect of IR or other therapies. 相似文献
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采用溶瘤腺病毒策略进行肝细胞癌基因治疗的思考 总被引:1,自引:0,他引:1
研制有效的盱细胞癌生物治疗制剂,作为现有治疗手段的有益补充或替代,具有重大的社会和经济效益。采用腺病毒溶瘤策略开展肝细胞癌的基因治疗研究是一个新的方向。章概述了此方向研究所面临的问题,并提出了一些设想。 相似文献
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构建miRNA.29a/c的重组腺病毒并观察其对膀胱癌T24细胞增殖能力的调控。以人全基因组DNA为模板,PcR扩增miR-29a、miR-29c,克隆至腺病毒穿梭载体pAdtrace.TO4-CMV。重组穿梭载体经pmeI线性化后与腺病毒骨架质粒pAdEasy-1共转化感受态大肠杆菌BJ5183,通过同源重组获得重组腺病毒质粒pAdEasy—1—miR-29a、pAdEasy—1.miR-29c,pacI线性化后转染HEK-293细胞,进行包装和扩增。实时荧光定量PcR检测感染腺病毒的膀胱癌T24细胞miR-29a、miR.29c的表达水平,并利用CCK.8实验检测细胞增殖能力。经DNA测序和限制性内切酶分析显示,重组腺病毒质粒pAdEasy—1—miR.29a、pAdEasy.1-miR-29c构建成功:感染腺病毒Ad—miR-29a和Ad—miR.29c后,经实时荧光定量PCR检测,膀胱癌细胞中miR.29a、miR.29c表达显著增高(P〈0.01);过表达miR.29a/c后的CCK.8实验显示,细胞增殖能力明显低于对照组伊〈0.05)。以上说明已成功构建miR.29a、miR.29c腺病毒,过表达miR.29a/c可抑制膀胱癌细胞的增殖。 相似文献
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Human CD59 Incorporation into Porcine Endogenous Retrovirus Particles: Implications for the Use of Transgenic Pigs for Xenotransplantation
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Daniel M. Takefman Gregory T. Spear Mohammed Saifuddin Carolyn A. Wilson 《Journal of virology》2002,76(4):1999-2002
Transgenic pigs have been engineered to express human CD59 (hCD59) in order to suppress hyperacute rejection of xenotransplants in human recipients. In this study, porcine endogenous retrovirus (PERV) was produced in a porcine cell line expressing hCD59 in order to examine the effect of this complement control protein on PERV neutralization by human sera. hCD59 was found to be incorporated into PERV particles produced from engineered ST-IOWA cells. PERV incorporation of hCD59 resulted in a dramatic inhibition of complement-mediated virolysis by human serum. However, incorporation of hCD59 had no effect on neutralization of PERV by human serum, as measured in infectivity assays. Our results suggest that the use of organs from hCD59 transgenic pigs will inhibit complement-mediated virolysis, but will not compromise the protective effects of human sera on the neutralization of PERV particles. 相似文献
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Veronica Rainone Cristina Martelli Luisa Ottobrini Mara Biasin Manuela Borelli Giovanni Lucignani Daria Trabattoni Mario Clerici 《PloS one》2016,11(1)
IntroductionDendritic cells play a key role as initiators of T-cell responses, and even if tumour antigen-loaded dendritic cells can induce anti-tumour responses, their efficacy has been questioned, suggesting a need to enhance immunization strategies.ResultsResults showed that antigen-loaded dendritic cells are characterized by a phenotypically semi-mature/mature profile and by the upregulation of genes involved in antigen presentation and T-cell priming. Activated dendritic cells stimulated T-cell proliferation and induced the production of high concentrations of IL-12p70 and IFN-γ but only low levels of IL-10, indicating their ability to elicit a TH1-immune response. Furthermore, administration of Antigen loaded-Dendritic Cells in MMTV-Ras mice evoked a strong anti-tumour response in vivo as demonstrated by a general activation of immunocompetent cells and the release of TH1 cytokines.ConclusionData herein could be useful in the design of antitumoral DC-based therapies, showing a specific activation of immune system against breast cancer. 相似文献
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目的构建并鉴定带有绿色荧光蛋白(green fluorescence protein,GFP)报告基因的人源CUL4A(hCUIAA)基因腺病毒表达载体Ad—hCUIAA—GFP,探求bCUIAA在PC-12细胞中的表达特点。方法扩增hCUIAA基因,并通过In—FusionPCR克隆技术构建穿梭质粒pDC315-EGFP—hCUIAA,利用AdMaxTM腺病毒包装系统将该穿梭质粒与腺病毒表达载体骨架质粒pBHGloxAEl,E3Cre共转染HEK293细胞,经GFP荧光检测和Western印迹检测确认hCUIAA的表达后,进一步通过病毒扩增及纯化得到hCUL4A重组腺病毒载体Ad—hCUIAA—GFP。将该载体转染Pc—12细胞,观察hCUIAA—GFP融合蛋白在Pc-12细胞中的表达情况。结果成功获得了较高滴度的腺病毒载体Ad—hCUIAA—GFP(1.6×10^12pfu/m1)。荧光检测表明,Ad—hCUIAA—GFP转染Pc-12细胞后72h内,病毒转染率随着时间和病毒转染滴度的增加而增加。DAPI细胞核荧光染色结果表明,hCUIAA—GFP的表达主要集中在细胞质部分。GFP荧光检测及Western印迹检测结果显示,hCUIAA—GFP在Pc-12细胞中的表达量随时间和病毒转染滴度的增加而增加。结论带GFP的hCUIAA重组腺病毒载体Ad—hCUIAA—GFP构建成功,掌握了其转染Pc-12细胞的最佳滴度及其在Pc-12细胞中的时空表达特点,为今后对hCUIAA在PC-12细胞中的功能性研究奠定了基础。 相似文献
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Li Liu Liang Liu Lai-Han Leung Austin J. Cooney Changyi Chen Todd K. Rosengart Yupo Ma Jianchang Yang 《The Journal of biological chemistry》2015,290(17):10599-10609
All-trans retinoic acid (ATRA) is a differentiation agent that revolutionized the treatment of acute promyelocytic leukemia. However, it has not been useful for other types of acute myeloid leukemia (AML). Here we explored the effect of SALL4, a stem cell factor, on ATRA-induced AML differentiation in both ATRA-sensitive and ATRA-resistant AML cells. Aberrant SALL4 expression has been found in nearly all human AML cases, whereas, in normal bone marrow and peripheral blood cells, its expression is only restricted to hematopoietic stem/progenitor cells. We reason that, in AMLs, SALL4 activation may prevent cell differentiation and/or protect self-renewal that is seen in normal hematopoietic stem/progenitor cells. Indeed, our studies show that ATRA-mediated myeloid differentiation can be largely blocked by exogenous expression of SALL4, whereas ATRA plus SALL4 knockdown causes significantly increased AML differentiation and cell death. Mechanistic studies indicate that SALL4 directly associates with retinoic acid receptor α and modulates ATRA target gene expression. SALL4 is shown to recruit lysine-specific histone demethylase 1 (LSD1) to target genes and alter the histone methylation status. Furthermore, coinhibition of LSD1 and SALL4 plus ATRA treatment exhibited the strongest anti-AML effect. These findings suggest that SALL4 plays an unfavorable role in ATRA-based regimes, highlighting an important aspect of leukemia therapy. 相似文献
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Zhen Sun Beier Luo Zhi-Heng Liu Dino Samartzis Zhongyang Liu Bo Gao Liangliang Huang Zhuo-Jing Luo 《International journal of biological sciences》2015,11(2):133-143
Introduction: Abnormal biomechanics plays a role in intervertebral disc degeneration. Adipose-derived stromal cells (ADSCs) have been implicated in disc integrity; however, their role in the setting of mechanical stimuli upon the disc''s nucleus pulposus (NP) remains unknown. As such, the present study aimed to evaluate the influence of ADSCs upon NP cells in compressive load culture.Methods: Human NP cells were cultured in compressive load at 3.0MPa for 48 hours with or without ADSCs co-culture (the ratio was 50:50). We used flow cytometry, live/dead staining and scanning electron microscopy (SEM) to evaluate cell death, and determined the expression of specific apoptotic pathways by characterizing the expression of activated caspases-3, -8 and -9. We further used real-time (RT-) PCR and immunostaining to determine the expression of the extracellular matrix (ECM), mediators of matrix degradation (e.g. MMPs, TIMPs and ADAMTSs), pro-inflammatory factors and NP cell phenotype markers.Results: ADSCs inhibited human NP cell apoptosis via suppression of activated caspase-9 and caspase-3. Furthermore, ADSCs protected NP cells from the degradative effects of compressive load by significantly up-regulating the expression of ECM genes (SOX9, COL2A1 and ACAN), tissue inhibitors of metalloproteinases (TIMPs) genes (TIMP-1 and TIMP-2) and cytokeratin 8 (CK8) protein expression. Alternatively, ADSCs showed protective effect by inhibiting compressive load mediated increase of matrix metalloproteinases (MMPs; MMP-3 and MMP-13), disintegrin and metalloproteinase with thrombospondin motifs (ADAMTSs; ADAMTS-1 and 5), and pro-inflammatory factors (IL-1beta, IL-6, TGF-beta1 and TNF-alpha).Conclusions: Our study is the first in vitro study assessing the impact of ADSCs on NP cells in an un-physiological mechanical stimulation culture environment. Our study noted that ADSCs protect compressive load induced NP cell death and degradation by inhibition of activated caspase-9 and -3 activity; regulating ECM and modulator genes, suppressing pro-inflammatory factors and preserving CK8. Consequently, the protective impact of ADSCs found in this study provides an essential understanding and expands our knowledge as to the utility of ADSCs therapy for intervertebral disc regeneration. 相似文献
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Jie Lu Kohei Yoshimura Koichi Goto Craig Lee Ken Hamura Ohyun Kwon Fuyuhiko Tamanoi 《PloS one》2015,10(9)
Small molecule inhibitors against protein geranylgeranyltransferase-I such as P61A6 have been shown to inhibit proliferation of a variety of human cancer cells and exhibit antitumor activity in mouse models. Development of these inhibitors could be dramatically accelerated by conferring tumor targeting and controlled release capability. As a first step towards this goal, we have encapsulated P61A6 into a new type of liposomes that open and release cargos only under low pH condition. These low pH-release type liposomes were prepared by adjusting the ratio of two types of phospholipid derivatives. Loading of geranylgeranyltransferase-I inhibitor (GGTI) generated liposomes with average diameter of 50–100 nm. GGTI release in solution was sharply dependent on pH values, only showing release at pH lower than 6. Release of cargos in a pH-dependent manner inside the cell was demonstrated by the use of a proton pump inhibitor Bafilomycin A1 that Increased lysosomal pH and inhibited the release of a dye carried in the pH-liposome. Delivery of GGTI to human pancreatic cancer cells was demonstrated by the inhibition of protein geranylgeranylation inside the cell and this effect was blocked by Bafilomycin A1. In addition, GGTI delivered by pH-liposomes induced proliferation inhibition, G1 cell cycle arrest that is associated with the expression of cell cycle regulator p21CIP1/WAF1. Proliferation inhibition was also observed with various lung cancer cell lines. Availability of nanoformulated GGTI opens up the possibility to combine with other types of inhibitors. To demonstrate this point, we combined the liposomal-GGTI with farnesyltransferase inhibitor (FTI) to inhibit K-Ras signaling in pancreatic cancer cells. Our results show that the activated K-Ras signaling in these cells can be effectively inhibited and that synergistic effect of the two drugs is observed. Our results suggest a new direction in the use of GGTI for cancer therapy. 相似文献