Trans-complementation of vector replication versus Coxsackie-adenovirus-receptor overexpression to improve transgene expression in poorly permissive cancer cells

Fechner, Henry; Wang, X; Wang, H; Jansen, A.; Pauschinger, Matthias; Scherübl, Hans; Bergelson, Jeffrey M.; Schultheiss, H.‐P.; Poller, Wolfgang

doi:10.1038/sj.gt.3301321

Cited by 79 publications

(70 citation statements)

References 39 publications

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“…The variable expression of CAR has been documented in many cancer types including glioma, melanoma, bladder cancer, rhabdomyosarcoma, neuroblastoma and ovarian cancer. [24][25][26][65][66][67][68][69] Furthermore, the downregulation of CAR may be associated with a more malignant phenotype. 70 Due to the variable expression of CAR on human primary cancer cells, the utility of Ad5 as a cancer gene therapy vector is compromised, limiting overall efficiency of cancer gene therapy including the use of CRAds.…”

Section: Discussionmentioning

confidence: 99%

Infectivity enhanced, hTERT promoter-based conditionally replicative adenoviruses are useful for SCLC treatment

et al. 2005

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Treatment of advanced small-cell lung cancer (SCLC) remains one of the major challenges in current medicine because of the high morbidity and mortality of the disease. Advanced stage lung cancer is refractory to conventional therapies and it also has an extremely poor prognosis. As a result, new therapeutic approaches are needed. Telomere maintenance to the regulation of replicative lifespan strongly implies that alterations in telomere biology play an important role during malignant transformation. Cancers that exhibit high levels of telomerase activity, such as all of the SCLC, were examined in a previous study. In this study, we turned the expression of human telomerase reverse transcriptase (hTERT) by tumors to a therapeutic advantage using a conditionally replication-competent adenovirus (CRAd) in which the expression of E1 (early region 1) is controlled by the hTERT promoter. This virus achieved good levels of viral replication in SCLC cells and induced a substantial anticancer effect in vitro and in vivo. As a further enhancement, the cancer cell killing effect was improved with a tropism modification of the virus to express the knob domain of Ad3 (serotype 3 adenovirus), and this improved infectivity for cancer cells. Conversely, the hTERT promoter has low activity in normal tissues, and the CRAd caused no damage to normal lung fibroblast cells. Since the telomerase activity is common in many types of cancers, these CRAds may be applicable to a wide range of tumors. We concluded that the use of hTERT promoterbased CRAds may be a potentially effective strategy for cancer treatment.

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Section: Discussionmentioning

confidence: 99%

Infectivity enhanced, hTERT promoter-based conditionally replicative adenoviruses are useful for SCLC treatment

et al. 2005

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“…A strong correlation between the expression of CAR on the cell surface and the level of transgene expression in photochemically treated cells was observed, suggesting that the absolute level of virus binding to the cell surface determines the level of transduction in photochemically treated cells, as reported by other groups for conventional adenoviral infection. [18][19][20] Blocking CAR with recombinant Ad5 fiber-knob resulted in a complete inhibition of gene expression, both in conventionally infected and in photochemically treated cells. These results strengthen the assumption that attachment to the cell surface is a key event for PCI-mediated transduction and suggest further that nonreceptor-mediated uptake pathways are not involved in the photochemical transduction pathway.…”

Section: Discussionmentioning

confidence: 99%

“…Using the RmcB anti-CAR monoclonal antibody, 13 we found that the cell lines used in this study expressed different levels of CAR ( Table 1) Fig 1b), as previously reported by other groups. [18][19][20] The relationship between the expression of CAR and the percentage of b-galactosidasepositive cells in photochemically treated cells showed an even better linear correlation (P ¼ .002 , Fig 1b), suggesting that virus binding to the cell surface is of fundamental importance also for photochemical transduction.…”

Section: Influence Of Car On Photochemical Transductionmentioning

confidence: 99%

PCI-enhanced adenoviral transduction employs the known uptake mechanism of adenoviral particles

et al. 2005

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The development of methods for efficient and specific delivery of therapeutic genes into target tissues is an important issue for further development of in vivo gene therapy. In the present study, the physical targeting technique, photochemical internalization (PCI), has been used together with adenovirus. The combination of PCI and adenoviral transduction has previously been shown to be favorable compared to adenovirus used alone, and the aim of this study was to verify the role of the adenoviral receptors and identify the uptake pathway used by adenoviral particles in photochemically treated cells. All examined cell lines showed augmented transduction efficiency after PCI-treatment, with a maximum of 13-fold increase in transgene expression compared to conventionally infected cells. Blocking of CAR induced a complete inhibition of PCI-enhanced transgene expression. However, photochemical treatment managed to enhance the transduction efficiency of the retargeted virus AdRGD-GFP showing also that the virus-CAR interaction is not vital for obtaining a photochemical effect on adenoviral transduction. Blocking the a V -integrins reduced the gene expression significantly in photochemically treated cells. Subjecting HeLa cells expressing negative mutant-dynamin to light treatment after infection gave no significant increase in gene transfer, while the gene transfer were enhanced seven-fold in cells with wild-type dynamin. Furthermore, chlorpromazine inhibited photochemical transduction in a dose-dependent manner, whereas Filipin III had no effect on the gene transfer. In summary, the data presented imply that adenoviral receptor binding is important and clathrin-mediated endocytosis is the predominant uptake mechanism for adenoviral particles in photochemically treated cells.

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“…The adenoviral vectors Ad5-CMVrtTA-M2, Ad5CMVluc, Ad5TREluc 23 and Ad.E (denoted earlier as Ad5TetO 7 SPB-E1A DpRB ) 22 have been described. All adenoviral constructs were tested for RCA contamination by PCR as described before 58 and the viral titers were determined using standard plaque assays on HEK293 cell.…”

Section: Construction Of Adenoviral Vectorsmentioning

confidence: 99%

“…58 Briefly, genomic plus episomal vector DNA were isolated by using the E.Z.N.A. s Tissue DNA Kit II (Peqlab Biotechnologie GmbH).…”

Section: Semiquantitative Pcrmentioning

confidence: 99%

Tamoxifen-regulated adenoviral E1A chimeras for the control of tumor selective oncolytic adenovirus replication in vitro and in vivo

et al. 2005

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Pharmacological control is a desirable safety feature of oncolytic adenoviruses (oAdV). It has recently been shown that oAdV replication may be controlled by drug-dependent transcriptional regulation of E1A expression. Here, we present a novel concept that relies on tamoxifen-dependent regulation of E1A activity through functional linkage to the mutated hormone-binding domain of the murine estrogen receptor (Mer). Four different E1A-Mer chimeras (ME, EM, E DNLS M, MEM) were constructed and inserted into the adenoviral genome under control of a lung-specific surfactant protein B promoter. The highest degree of regulation in vitro was seen for the corresponding oAdVs Ad.E DNLS M and Ad.MEM, which exhibited an up to 100-fold higher oAdV replication in the presence as compared with the absence of 4-OH-tamoxifen. Moreover, destruction of nontarget cells was six-and 13-fold reduced for Ad.E DNLS M and Ad.MEM, respectively, as compared with Ad.E. Further investigations supported tamoxifen-dependent regulation of Ad.E DNLS M and Ad.MEM in vivo. Induction of Ad.E DNLS M inhibited growth of H441 lung tumors as efficient as a control oAdV expressing E1A. E DNLS M and the MEM chimeras can be easily inserted into a single vector genome, which extends their application to existing oAdVs and strongly facilitates in vivo application.

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Trans-complementation of vector replication versus Coxsackie-adenovirus-receptor overexpression to improve transgene expression in poorly permissive cancer cells

Cited by 79 publications

References 39 publications

Infectivity enhanced, hTERT promoter-based conditionally replicative adenoviruses are useful for SCLC treatment

Infectivity enhanced, hTERT promoter-based conditionally replicative adenoviruses are useful for SCLC treatment

PCI-enhanced adenoviral transduction employs the known uptake mechanism of adenoviral particles

Tamoxifen-regulated adenoviral E1A chimeras for the control of tumor selective oncolytic adenovirus replication in vitro and in vivo

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