Adenoviral vector which delivers FasL–GFP fusion protein regulated by the tet-inducible expression system

Rubinchik, Semyon; Ding, Rongxian; Qiu, An-Chun; Zhang, F; Dong, Jian-Yun

doi:10.1038/sj.gt.3301172

Cited by 60 publications

(69 citation statements)

References 47 publications

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“…Development and characterization of 293CrmA has been previously described. 31 All cell lines were maintained in DMEM supplemented with 10% cosmic calf serum (CCS; HyClone) and 1% penicillin/streptomycin. For DNA transfections, 5 Â 10 5 cells/well were seeded in six-well plates and transfected 24 hours later using LipofectAMINE (Gibco BRL) according to the manufacturer's instructions.…”

Section: Cell Linesmentioning

confidence: 99%

“…31,40,41 Vectors expressing TRAIL or both FasL-GFP and TRAIL were prepared using similar methods. All vectors were banded twice on CsCl gradients and desalted twice with a PD-10 size exclusion column (Amersham Scientific) into HBS (HEPES-buffered saline) þ 5% glycerol, and stored at À701C.…”

Section: Infection Of Cultured Cells With Recombinant Ad Vectors In Vmentioning

confidence: 99%

“…Several published reports describe the construction and analysis of replication-defective recombinant Ad vectors that deliver FasL 25,27,[30][31][32][33][34] or TRAIL. 35,36 These vectors have not been uniformly successful against tumors established in animal models.…”

mentioning

confidence: 99%

“…Recently, we have reported development of a murine Fas ligand-green fluorescent protein (FasL-GFP) fusion protein, which we have shown to be stable, to have high apoptosis-inducing activity, and to be resistant to the metalloproteinase activity that in many tumor isolates cleaves off the extracytoplasmic C-terminus of human FasL. 31 We have put this protein under the control of a Tet-regulated promoter and have cloned both the FasL-GFP expression cassette and the Tet-regulated transcriptional activator protein expression cassette (using the CMV promoter) into a complex recombinant Ad5 vector. This combination resulted in high expression levels of the FasL-GFP protein under the induced conditions in all cultured cell lines and tumor isolates tested, and in the effective induction of apoptosis in several prostate cancer cell lines previously described as FasL-resistant.…”

mentioning

confidence: 99%

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Enhanced apoptosis of glioma cell lines is achieved by co-delivering FasL-GFP and TRAIL with a complex Ad5 vector

Rubinchik

Woraratanadharm

et al. 2003

Cancer Gene Ther

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Brain tumors (BTs) are among the most malignant forms of human cancer. Unfortunately, current treatments are often ineffective and produce severe side effects. Cytotoxic gene therapy is an alternative treatment strategy, with the potential advantages of reduced toxicity to normal brain tissue. Apoptosis-inducing ''death ligands'' Fas ligand and TNF-related apoptosis-inducing ligand (TRAIL) are genes with substantial cytotoxic activity in susceptible tumor cells. Here, we compared the effectiveness of Ad vectormediated delivery of Fas ligand-green fluorescent protein (FasL-GFP) fusion protein, human TRAIL, and both genes simultaneously. We examined a panel of 13 cell lines (eight derived from primary isolates) for susceptibility to Ad5-based vector infection and for sensitivity to FasL-and TRAIL-mediated apoptosis. All cell lines were efficiently transduced, but, as expected, varied in their sensitivity to ligand-induced apoptosis. Generally, sensitivity to FasL-GFP correlated with cell surface FasR levels, but no such correlation was seen for TRAIL and its functional receptors, DR4 and DR5. The vector expressing both FasL-GFP and TRAIL was more effective than either of the single-gene vectors at comparable transduction levels, and it was effective against a broader range of cell lines. In five cell lines, coexpression resulted in apoptosis levels greater than those predicted for strictly additive activity of the two death ligands. We believe that Ad vector-mediated delivery of multiple death ligands may be developed as a potential BT therapy, either alone or in conjunction with surgical resection of the primary tumor.

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Section: Cell Linesmentioning

confidence: 99%

Section: Infection Of Cultured Cells With Recombinant Ad Vectors In Vmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Enhanced apoptosis of glioma cell lines is achieved by co-delivering FasL-GFP and TRAIL with a complex Ad5 vector

Rubinchik

Woraratanadharm

et al. 2003

Cancer Gene Ther

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“…RAdhCMV-mFasL was verified to be free of RCA contamination by polymerase chain reaction, due to toxicity of mFasL expression on HeLa cells. 32 Aliquots of vectors used were found to be negative for RCA at levels used in the experiments.…”

Section: Recombinant Adenovirusesmentioning

confidence: 99%

Adenovirus-mediated expression of HSV1-TK or Fas ligand induces cell death in primary human glioma-derived cell cultures that are resistant to the chemotherapeutic agent CCNU

et al. 2001

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An increased transforming growth factor β receptor type I:Type II ratio contributes to elevated collagen protein synthesis that is resistant to inhibition via a kinase‐deficient transforming growth factor β receptor type II in scleroderma

Pannu¹,

Gore-Hyer²,

Yamanaka³

et al. 2004

Arthritis & Rheumatism

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Objective. Aberrant transforming growth factor ␤ (TGF␤) signaling has been implicated in the pathogenesis of scleroderma (systemic sclerosis [SSc]), but the contribution of specific components in this pathway to SSc fibroblast phenotype remains unclear. This study was undertaken to delineate the role of TGF␤ receptor type I (TGF␤RI) and TGF␤RII in collagen overexpression by SSc fibroblasts.Methods. Primary dermal fibroblasts from SSc patients and healthy adults were studied (n ‫؍‬ 10 matched pairs). Adenoviral vectors were generated for TGF␤RI (AdTGF␤RI), TGF␤RII (AdTGF␤RII), and kinase-deficient TGF␤RII (Ad⌬kRII). TGF␤RI basal protein levels were analyzed by 35 S-methionine labeling/ immunoprecipitation and immunohistochemistry. Type I collagen and TGF␤RII basal protein levels were analyzed by Western blot and newly secreted collagen by 3 H-proline incorporation assay.Results. Analysis of endogenous TGF␤RI and TGF␤RII protein levels revealed that SSc TGF␤RI levels were increased 1.7-fold (P ‫؍‬ 0.008; n ‫؍‬ 7) compared with levels in healthy controls, while TGF␤RII levels were decreased by 30% (P ‫؍‬ 0.03; n ‫؍‬ 7). This increased TGF␤RI:TGF␤RII ratio correlated with SSc collagen overexpression. To determine the consequences of altered TGF␤RI:TGF␤RII ratio on collagen expression, healthy fibroblasts were transduced with AdTGF␤RI or AdTGF␤RII. Forced expression of TGF␤RI in the range corresponding to elevated SSc TGF␤RI levels increased basal collagen expression in a dose-dependent manner, while similar TGF␤RII overexpression had no effect, although transduction of fibroblasts at higher multiplicities of infection led to a marked reduction of basal collagen levels. Blockade of TGF␤ signaling via Ad⌬kRII resulted in ϳ50% inhibition of basal collagen levels in healthy fibroblasts and in 5 of 9 SSc cell lines. A subset of SSc fibroblasts (4 of 9 cell lines) was resistant to this treatment. SSc fibroblasts with the highest levels of TGF␤RI were the least responsive to collagen inhibition via ⌬kRII.Conclusion. This study indicates that an increased TGF␤RI:TGF␤RII ratio may underlie aberrant TGF␤ signaling in SSc and contribute to elevated basal collagen production, which is insensitive to TGF␤ signaling blockade via ⌬kRII.Scleroderma (systemic sclerosis [SSc]) is characterized by excessive deposition of extracellular matrix

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Adenoviral vector which delivers FasL–GFP fusion protein regulated by the tet-inducible expression system

Cited by 60 publications

References 47 publications

Enhanced apoptosis of glioma cell lines is achieved by co-delivering FasL-GFP and TRAIL with a complex Ad5 vector

Enhanced apoptosis of glioma cell lines is achieved by co-delivering FasL-GFP and TRAIL with a complex Ad5 vector

Adenovirus-mediated expression of HSV1-TK or Fas ligand induces cell death in primary human glioma-derived cell cultures that are resistant to the chemotherapeutic agent CCNU

An increased transforming growth factor β receptor type I:Type II ratio contributes to elevated collagen protein synthesis that is resistant to inhibition via a kinase‐deficient transforming growth factor β receptor type II in scleroderma

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