Herpes simplex virus thymidine kinase/ganciclovir-induced apoptosis involves ligand-independent death receptor aggregation and activation of caspases

Beltinger, Christian; Fulda, Simone; Kammertoens, Thomas; Meyer, Éric; Uckert, Wolfgang; Debatin, Klaus‐Michael

doi:10.1073/pnas.96.15.8699

Cited by 163 publications

(134 citation statements)

References 32 publications

Supporting

Mentioning

124

Contrasting

Unclassified

Order By: Relevance

“…2 Reasoning that TRAIL -a death -inducing ligand, which also interacts with members of the TNF receptor superfamily -may have similar effects but may be better suited for tumor therapy because of its preferential action on tumor cells, we investigated TK /GCV -induced sensitization to rhTRAIL in SH -EP neuroblastoma cells.…”

Section: Resultsmentioning

confidence: 99%

“…In SH -EP neuroblastoma cells, TK /GCV induces caspase activation and TK -induced apoptosis is caspase-dependent. 2 TRAIL -mediated apoptosis is also caspase -dependent. 31 Thus, concerted caspase activation by TK /GCV and rhTRAIL may reduce the dose of either stimulus needed to activate caspases because these are activated already to a certain degree by the other stimulus.…”

Section: Discussionmentioning

confidence: 99%

“…Cells were cultured in RPMI 1640 medium ( Life Technologies, Eggenstein, Germany ) supplemented with 10% heat -inactivated FCS (Conco, Wiesbaden, Germany ), 10 mM HEPES, pH 7.3 (Biochrom, Berlin, Germany ), 100 U /mL penicillin (Life Technologies ), 100 g /mL streptomycin ( Life Technologies ), 2 mM L -glutamine (Biochrom ), 0.5 mg /mL G418 (Life Technologies), or 0.5 mg /mL hygromycin ( Life Technologies ), as previously described. 2,14,44 Clonogenicity assay 2Â10 4 TK + or TK À cells per well were plated into 12-well plates, allowed to attach overnight, and then treated with GCV in indicated concentrations for 4 days. Cells still adherent were trypsinized and pooled with the cells already detached.…”

Section: Cellsmentioning

confidence: 99%

“…In many tumor cell types, apoptosis is the prevalent response to GCV of both TK -bearing cells 2,12,18,28,51 and of bystander cells. 5,12,19 Nonapoptotic, e.g., necrotic, cell death was reported in some tumor cell types such as B16 melanoma after TK /GCV therapy.…”

mentioning

confidence: 99%

“…1,37,44,49 TK / GCV -induced apoptosis in SH -EP neuroblastoma cells is triggered by ligand -independent aggregation of CD95, which recruits the adaptor molecule FADD and caspase -8, forming a death -inducing signaling complex ( DISC ). 2 Caspase-8 thus becomes activated and in turn activates downstream effector caspases by proteolytic cleavage, leading to apoptosis. TK /GCV also affects the second mitochondrial pathway of apoptosis induction.…”

mentioning

confidence: 99%

See 4 more Smart Citations

TRAIL enhances thymidine kinase/ganciclovir gene therapy of neuroblastoma cells

et al. 2002

Self Cite

View full text Add to dashboard Cite

The clinical benefit of suicide gene therapy of tumors has been marginal, mostly due to the low gene transfer efficiency in vivo. The death -inducing ligand, TRAIL, effectively kills many tumor cell types, while sparing most normal tissues. We hypothesized that TRAIL may enhance HSV thymidine kinase / ganciclovir ( TK / GCV ) gene therapy of tumor cells by augmenting both target and bystander cell kill. Human SH -EP neuroblastoma cells expressing TK as well as bystander cells were effectively killed by apoptosis, and their clonogenicity was ablated following GCV. Human TRAIL enhanced TK / GCV -induced cell death and decreased clonogenicity of TK -expressing cells and also of bystander cells. Cooperation between TRAIL and TK / GCV depended both on caspase activation and on mitochondrial apoptogenic function because both the broad -spectrum caspase inhibitor zVAD.fmk and overexpression of Bcl -2 decreased enhancement of cell kill by TRAIL. Facilitation of TRAIL signalling by up -regulation of TRAIL receptors did not contribute to enhancement because cell surface expression of the agonistic TRAIL receptors 1 and 2 was not increased by TK / GCV. In conclusion, the concerted activation of caspases and the mitochondrial amplification of caspase activation by TK / GCV may explain the cooperative effect of TK / GCV and TRAIL on the kill of neuroblastoma cells. Because combined treatment also augmented the bystander cell kill, the addition of TRAIL may increase the efficacy of TK / GCV gene therapy of neuroblastoma.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Cellsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

TRAIL enhances thymidine kinase/ganciclovir gene therapy of neuroblastoma cells

et al. 2002

Self Cite

View full text Add to dashboard Cite

show abstract

Nucleoside binding site ofHerpes simplex type 1 thymidine kinase analyzed by X-ray crystallography

Vogt¹,

Perozzo²,

Pautsch³

et al. 2000

Proteins

View full text Add to dashboard Cite

The crystal structures of the full‐length Herpes simplex virus type 1 thymidine kinase in its unligated form and in a complex with an adenine analogue have been determined at 1.9 Å resolution. The unligated enzyme contains four water molecules in the thymidine pocket and reveals a small induced fit on substrate binding. The structure of the ligated enzyme shows for the first time a bound adenine analogue after numerous complexes with thymine and guanine analogues have been reported. The adenine analogue constitutes a new lead compound for enzyme‐prodrug gene therapy. In addition, the structure of mutant Q125N modifying the binding site of the natural substrate thymidine in complex with this substrate has been established at 2.5 Å resolution. It reveals that neither the binding mode of thymidine nor the polypeptide backbone conformation is altered, except that the two major hydrogen bonds to thymidine are replaced by a single water‐mediated hydrogen bond, which improves the relative acceptance of the prodrugs aciclovir and ganciclovir compared with the natural substrate. Accordingly, the mutant structure represents a first step toward improving the virus‐directed enzyme‐prodrug gene therapy by enzyme engineering. Proteins 2000;41:545–553. © 2000 Wiley‐Liss, Inc.

show abstract

Gene directed enzyme/prodrug therapy of cancer: Historical appraisal and future prospectives

2001

View full text Add to dashboard Cite

Gene therapy of cancer is a novel approach with the potential to selectively eradicate tumour cells, whilst sparing normal tissue from damage. In particular, gene-directed enzyme prodrug therapy (GDEPT) is based on the delivery of a gene that encodes an enzyme which is non-toxic per se, but is able to convert a prodrug into a potent cytotoxin. Several GDEPT systems have been investigated so far, demonstrating effectiveness in both tissue culture and animal models. Based on these encouraging results, phase I/II clinical trials have been performed and are still ongoing. The aim of this review is to summarise the progress made in the design and application of GDEPT strategies. The most widely used enzyme/prodrug combinations already in clinical trials (e.g., herpes simplex 1 virus thymidine kinase/ganciclovir and cytosine deaminase/5-¯uorocytosine), as well as novel approaches (carboxypeptidase G2/CMDA, horseradish peroxidase/indole-3-acetic acid) are described, with a particular attention to translational research and early clinical results.

show abstract

Herpes simplex virus thymidine kinase/ganciclovir-induced apoptosis involves ligand-independent death receptor aggregation and activation of caspases

Abstract: ABSTRACT

Cited by 163 publications

References 32 publications

TRAIL enhances thymidine kinase/ganciclovir gene therapy of neuroblastoma cells

TRAIL enhances thymidine kinase/ganciclovir gene therapy of neuroblastoma cells

Nucleoside binding site ofHerpes simplex type 1 thymidine kinase analyzed by X-ray crystallography

Gene directed enzyme/prodrug therapy of cancer: Historical appraisal and future prospectives

Contact Info

Product

Resources

About