Specific tumor-cell killing with adenovirus vectors containing the apoptin gene

Pietersen, Alexandra M; Eb, M.M. van der; Rademaker, Hendrik J.; Wollenberg, Diana J.M. van den; Rabelink, Martijn J. W. E.; Kuppen, Peter J. K.; Dierendonck, Jan Hein van; Ormondt, H. van; Masman, D; Velde, Cornelis J.H. van de; Eb, A. J. Van Der; Hoeben, Rob C.; Noteborn, Mathieu H. M.

doi:10.1038/sj.gt.3300876

Cited by 90 publications

(73 citation statements)

References 36 publications

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“…ASGPRmediated transfer of the Apoptin gene does not adversely affect its ability to induce tumor-specific apoptosis. [3][4][5][6][7][8][9][10] PCR amplification revealed that, after injection of Asor-Apoptin complex into tumor-bearing nude mice via the tail vein, Apoptin gene existed stably and specifically in the liver and transplanted hepatocarcinoma tumor cells, indicating that the Asor-Apoptin complex was stably transported through the blood stream with hepatic targeting specificity. A single systemic treatment with Asor-Apoptin resulted already in a huge extent of damage to the hepatocarcinoma architecture.…”

Section: Discussionmentioning

confidence: 99%

“…The fixed cells were then washed in ice-cold PBS (containing 2 mM MgCl 2 ), and incubated in 3 ml of reaction mix (1 mg/ml X-gal (Roche, Shanghai, China), 5 mM potassium ferrocyanide, 5 mM potassium ferricyanide, 2 mM MgCl 2 in PBS) at 371C for 4-16 h. 8 To determine apoptosis induction, Tdt-mediated dUTP nick-end labeling (TUNEL) was performed by use of in situ cell death detection kit as described previously. 8,9 Animals and in situ hepatocarcinoma model Five-to six-week-old BALB/c nude mice with bodyweight of 16-22 g were provided by the Laboratory Animal Center of Tongji Medical College, Huazhong University of Science and Technology (PR China). The nude mice were kept at standard laboratory conditions of alternating 12-h periods of light and darkness and a standard laboratory diet.…”

Section: Preparation Of Asor-pll-dna Complexesmentioning

confidence: 99%

“…1,2 At present, anticancer therapies are limited by side toxicity to normal tissues and the occurrence of therapy-resistant tumor cells. 2 The viral death effector Apoptin protein [3][4][5][6] induces apoptosis in various hepatocarcinoma-derived cells, [7][8][9] but not in human primary hepatocytes. 10 Van der Eb et al 11 demonstrated that adenovirus-mediated intratumoral transfer and expression of the Apoptin gene resulted in regression or complete remission of human hepatocarcinomas grown as xenografts in immune-deficient mice.…”

Section: Introductionmentioning

confidence: 99%

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Inhibition of hepatocarcinoma by systemic delivery of Apoptin gene via the hepatic asialoglycoprotein receptor

Sun

et al. 2006

Cancer Gene Ther

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Specificity is a prerequisite for systemic gene therapy of hepatocarcinoma. In vitro, the tumor-specific viral death effector Apoptin selectively induces apoptosis in malignant hepatic cells. Intratumoral treatment of xenografted subcutaneous hepatomas with Apoptin results in tumor regression. Here, we report a systemic delivery vehicle containing the Apoptin gene linked to asialoglycoprotein (Asor), which targets asialoglycoprotein receptor (ASGPR) present only on the surface of hepatocytes. In vitro, the protein-DNA complex Asor-Apoptin induced apoptosis in HepG2 hepatocarcinoma cells but not in normal L-02 hepatocytes. Non-hepatocyte-derived tumorigenic human A549 cells lacking the membrane ASGPR were not affected by Asor-Apoptin. In vivo systemic delivery of Asor-Apoptin via the tail vein into mice bearing in situ hepatocarcinoma resulted in specific and efficient distribution of Apoptin in both hepatocarcinoma cells and normal hepatocytes. Five days after injection of Asor-Apoptin, the in situ hepatocarcinomas showed significant signs of regression, whereas the surrounding normal hepatocytes did not. Systemically delivered Asor-LacZ expressing non-apoptotic LacZ gene did not inhibit tumor growth. Our data reveal that systemic delivery of Asor-Apoptin specifically induces apoptosis in malignant hepatocytes and thus constitutes a powerful and safe therapeutics against hepatocarcinomas.

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Section: Discussionmentioning

confidence: 99%

Section: Preparation Of Asor-pll-dna Complexesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Inhibition of hepatocarcinoma by systemic delivery of Apoptin gene via the hepatic asialoglycoprotein receptor

Sun

et al. 2006

Cancer Gene Ther

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“…Thus, in addition to results from other gene therapy studies, 28 our data provide further evidence that alternative strategies are needed to improve the efficacy of in vivo gene therapy for tumors expressing wild -type p53 and mutated bax. Selective induction of apoptosis using transfer of proapoptotic genes like bax, 62 apoptin, 63 or caspases 64,65 directly into tumor cells in vivo appears to be a promising treatment strategy. However, a therapeutic effect using this approach requires intact downstream signal transduction pathways.…”

Section: Discussionmentioning

confidence: 99%

Adenovirus-mediated gene transfer of P16INK4/CDKN2 into bax-negative colon cancer cells induces apoptosis and tumor regression in vivo

et al. 2002

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The tumor -suppressor gene p16INK4 / CDKN2 ( p16 ) is a cyclin -dependent kinase ( cdk ) inhibitor and important cell cycle regulator. Here, we show that adenovirus -mediated gene transfer of p16 ( AdCMV.p16 ) into colon cancer cells induces uncoupling of S phase and mitosis and subsequently apoptosis. Flow cytometric analysis revealed that cells infected with AdCMV.p16 showed an initial G2 -like arrest followed by S phase without intervening mitosis ( DNA >4N ). Using microscopic analysis, deformed polyploid cells were detectable only in cells infected with AdCMV.p16 but not in control -infected cells. Subsequently, AdCMV.p16 -infected polyploid cells underwent apoptosis, as assessed by AnnexinV staining and DNA fragmentation, suggesting that cell cycle dysregulation is upstream of the onset of apoptosis. Treatment of mice with subcutaneously transplanted tumors of colorectal cancer cells with AdCMV.p16 but not AdCMV.p53 resulted in significantly reduced tumor volume and prolonged survival. Using an orthotopic model of liver metastasis, we observed both reduced local tumor growth and secondary intrahepatic metastasis after AdCMV.p16 treatment. Importantly, induction of apoptosis in vitro and reduction of tumor growth in vivo by p16 was p53 -as well as bax -independent because identical results were obtained using cancer cells, either wild type or mutant for p53 or bax. The studies suggest that an AdCMV.p16 -based treatment may be especially effective in patients with bax -negative colon cancer where overexpression of p53 appears not to be of therapeutic value.

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“…Amplification, propagation, and screening for replicationcompetent adenovirus were performed as described. 52,53 Creation of immune 'stealth' genes M Ossevoort et al Plaque assays were performed essentially as described. 51 Briefly, adenovirus stocks were serially diluted in 1 ml DMEM/2% HS and added to near-confluent 911 cells in six-well plates.…”

Section: Adenoviral Vector Constructionmentioning

confidence: 99%

Creation of immune ‘stealth’ genes for gene therapy through fusion with the Gly-Ala repeat of EBNA-1

et al. 2003

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A major obstacle in gene-therapy protocols is T-cellmediated destruction of transgene-expressing cells. Therefore new approaches are needed to prevent rapid clearance of transduced cells. We exploited the Gly-Ala repeat (GAr) domain of the Epstein-Barr virus nuclear antigen-1, since the GAr prevents cytotoxic T-lymphocyte-epitope generation. Here we show that three different enzymes (viz. the E. coli LacZ gene encoded b-galactosidase, firefly luciferase, and HSV1 thymidine kinase) fused with the GAr retained their function. Moreover, linking GAr with b-galactosidase successfully prevented recognition of GAr-LacZ-expressing cells by b-galactosidase-specific CTL. Nonetheless, vaccination with a GAr-LacZ adenovirus or with an allogeneic cell line expressing GAr-LacZ resulted in the induction of b-galspecific CTL. This demonstrates that the GAr domain does not inhibit crosspresentation of antigens, but only affects breakdown of endogenously synthesized proteins. These data demonstrate how the GAr domain can be exploited to create immuno'stealth' genes by hiding transgene products from CTL-mediated immune attack.

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Specific tumor-cell killing with adenovirus vectors containing the apoptin gene

Cited by 90 publications

References 36 publications

Inhibition of hepatocarcinoma by systemic delivery of Apoptin gene via the hepatic asialoglycoprotein receptor

Inhibition of hepatocarcinoma by systemic delivery of Apoptin gene via the hepatic asialoglycoprotein receptor

Adenovirus-mediated gene transfer of P16INK4/CDKN2 into bax-negative colon cancer cells induces apoptosis and tumor regression in vivo

Creation of immune ‘stealth’ genes for gene therapy through fusion with the Gly-Ala repeat of EBNA-1

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