Glioma/glioblastoma-specific adenoviral gene expression using the nestin gene regulator

Kurihara, Hideyuki; Zama, Akira; Tamura, Masahito; Takeda, Jun; Sasaki, Tatsuya; Takeuchi, Toshiyuki

doi:10.1038/sj.gt.3301129

Cited by 43 publications

(25 citation statements)

References 23 publications

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“…11 To make chimeric nestin regulators, we cut out the 2.1 kb nestin basic promoter (NP) region by Xmnl, as described before. 8 Then we cut out the 1.3 kb first intron (1i) DNA fragment by BamHI and AccI, and the 1.8 kb second intron (2i) fragment by BstYI. The 1i DNA fragment contains the 967 bp first intron, the 283 bp first exon, and the 37 bp second partial exon.…”

Section: Luciferase Assaymentioning

confidence: 99%

“…16 The antibody to human nestin was prepared in rabbits as described previously. 8 We obtained mouse monoclonal antibodies to human von Willebrand factor (vWF) and human Ki-67 (MIB-1) from DAKO (Glostrup, Denmark), mouse monoclonal antibodies to KDR and proliferating cell nuclear antigen (PCNA) from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA), and mouse monoclonal antibody to human CD34 from Nichirei (Tokyo, Japan).…”

Section: Morphological Studiesmentioning

confidence: 99%

“…For nestin-positive control cells, we used the U251 human glioblastoma cell line. 8 A tube formation assay with HUVECs was performed on Matrigel layer, 15 which is a crude extract of basal membrane from mouse Engelbreth-HolmSwarm tumors. Briefly, Matrigel (400 ml) (Collaborative Biomedical Products, Medford, MA, USA) was placed on a 3.0 cm culture plate and left for polymerization at 371C for 1 h. HUVECs were seeded at 3 Â 10 5 cells/plate, and cultured for an indicated time to form a tubular network.…”

Section: Cell Culturementioning

confidence: 99%

“…2,11 The neuroepitheliumspecific regulator, consisting of the second intron before the 2.1 kb-long 5 0 UP region of the gene, drove LacZ expression in parallel with the extent of mRNA expression in seven human glioma cell lines. 8 However, in terms of nestin expression in angiogenic endothelium, a regulatory region has not yet been identified.…”

mentioning

confidence: 99%

“…6 Furthermore, significant levels of nestin have been detected in human gliomas. 7,8 Recently, we re-evaluated the nestin expression in 71 cases of brain tumors including 40 cases of gliomas that ranged from low to high malignancy grades. 9 Nestin was highly immunostained in gliomas with a higher malignant grade.…”

mentioning

confidence: 99%

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Angiogenic endothelium-specific nestin expression is enhanced by the first intron of the nestin gene

Aihara

Sugawara

Torii

et al. 2004

Laboratory Investigation

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Nestin is a member of intermediate filaments abundantly expressed in neural stem cells and glioblastomas. The nestin gene has four exons and three introns, and neural cell-specific expression is regulated by the second intron. We previously reported that nestin was invariably detected in the tumor endothelium in gliomas even though tumor cells were negative for nestin. In the present study, we further confirmed nestin immunostaining in tumor endothelium of a variety of common cancers, including lung, stomach, colon, and cervical carcinomas. We examined an endothelium-specific regulator using human umbilical vein endothelial cells (HUVECs) and human glioblastoma-derived U251 cells. In a luciferase reporter assay, the first intron plus 5 0 upstream promoter (5 0 UP) gave the highest activity, followed by 5 0 UP, and the second intron plus 5 0 UP. However, the assay values were much lower by HUVEC extracts than by U251 cell extracts. Although green fluorescent protein expression was positive over all U251 cells under either the first intron, second intron, or ubiquitously active CAG promoter, the fluorescence in HUVECs was limited to a few cells even under the first intron. This difference came from the growth feature of HUVECs which exhibit growth arrest by contact inhibition. We found that the nestin expression was specific to proliferative endothelium, by using proliferation markers in hemangioblastomas and in situ hybridization. Using an endothelial tube formation assay, tyrosine kinase domain-deleted VEGF receptor KDR effectively abolished the tube formation under the first intron. We suggest that the nestin expression in tumor endothelium is enhanced by the first intron.

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Section: Luciferase Assaymentioning

confidence: 99%

Section: Morphological Studiesmentioning

confidence: 99%

Section: Cell Culturementioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Angiogenic endothelium-specific nestin expression is enhanced by the first intron of the nestin gene

Aihara

Sugawara

Torii

et al. 2004

Laboratory Investigation

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Hybrid vector designs to control the delivery, fate and expression of transgenes

Lam

Breakefield

2000

J. Gene Med.

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One of the greatest challenges to gene therapy is the targetting of gene delivery selectively to the sites of disease and regulation of transgene expression without adverse effects. Ultimately, the successful realization of these goals is dependent upon improvements in vector design. Over the years, viral vector design has progressed from various types of replication-defective viral mutants to replication-conditioned viruses and, more recently, to 'gutted' and hybrid vectors, which have, respectively, eliminated expression of non-relevant or toxic viral genes and incorporated desired elements of different viruses so as to increase the efficacy of gene delivery in vivo. This review will focus on the different viral and cellular elements which have been incorporated into virus vectors to: improve transduction efficiencies; alter the entry specificity of virions; control the fate of transgenes in the host cells; and regulate transgene expression.

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CD46 on glial cells can function as a receptor for viral glycoprotein‐mediated cell–cell fusion

Cassiani-Ingoni

Greenstone

Donati

et al. 2005

Glia

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Membrane cofactor protein (CD46) is a regulator of complement activation that also serves as the entry receptor for human herpes virus 6 (HHV-6) and measles virus (MV) into human cells. While it is clear that oligodendrocytes and astrocytes are cell types commonly infected by these viruses, it is unclear whether oligodendrocytes express CD46, or which are the cellular mechanisms underlying the infection. We show that adult oligodendrocytes, as well as astrocytes and microglial cells, express CD46 on the cellular surface. Moreover, we employed a quantitative fusion assay to demonstrate that HHV-6A infection of T lymphocytes enables cell-cell fusion of these cells to astrocytes or to oligodendroglial cells. This fusion is mediated by the interaction between viral glycoproteins expressed on the membrane of the infected cells and CD46 on the glial targets, and is also observed using cells expressing recombinant MV glycoproteins. These data suggest a mechanism that involves cell-cell fusion by which certain viruses could spread the infection from the periphery to the cells in the nervous system.

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Glioma/glioblastoma-specific adenoviral gene expression using the nestin gene regulator

Cited by 43 publications

References 23 publications

Angiogenic endothelium-specific nestin expression is enhanced by the first intron of the nestin gene

Angiogenic endothelium-specific nestin expression is enhanced by the first intron of the nestin gene

Hybrid vector designs to control the delivery, fate and expression of transgenes

CD46 on glial cells can function as a receptor for viral glycoprotein‐mediated cell–cell fusion

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