Development of an inducible suicide gene system based on human caspase 8

Carlotti, Françoise; Zaldumbide, Arnaud; Martin, Patrick; Boulukos, Kim E.; Hoeben, Rob C.; Pognonec, Philippe

doi:10.1038/sj.cgt.7700825

Cited by 39 publications

(36 citation statements)

References 52 publications

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“…One avenue to circumvent this problem is to develop new 'human' suicide genes. 9,37,38 As an alternative approach to evade the antigenspecific immune response, we evaluated the use of the GAr region of the nuclear antigen 1 (EBNA-1) of the EBV to protect HSV-TK-derived peptides from being presented. Here we provide evidence that fusing the GAr region of the EBNA-1 to HSV-TK reduces the presentation of transgene-product-derived peptides in vitro.…”

Section: Discussionmentioning

confidence: 99%

Characterization of an immuno ‘stealth’ derivative of the herpes simplex virus thymidine-kinase gene

et al. 2006

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The cellular immune response against transgene-encoded neoantigens is a potential hurdle in gene therapy applications where long-term expression of transgenes is desired. Here a new optimized derivative of the herpes simplex virus 1-thymidine-kinase (HSV1-TK) gene is described. The HSV-TK gene is frequently used in experimental studies on gene-directed enzyme prodrug therapy. In the optimized gene, the HSV-TK coding region is fused with the codons for the Gly-Ala repeat of the Epstein-Barr virus nuclear-antigen 1 to prevent proteasomal degradation of the HSV-TK. To measure the protective effect in vitro, a model cytotoxic T lymphocyte epitope derived from the ovalbumin was inserted in the TK. Cells expressing the GAr-modified TK do not present TKderived peptides in the major histocompatibility complex. Furthermore, conservative nucleotide substitutions were introduced, which prevent splicing, as well as mutations that render the TK-expressing cells more sensitive to ganciclovir (GCV). The GAr HSV-TK fusion protein is fully functional in vitro. This HSV-TK gene may be especially useful in those gene therapy applications where an immune response against the transgene-encoded product would frustrate the treatment.

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Section: Discussionmentioning

confidence: 99%

Characterization of an immuno ‘stealth’ derivative of the herpes simplex virus thymidine-kinase gene

et al. 2006

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“…were induced to go into apoptosis by expressing the FKC8 gene, which was activated after AP20187 treatment (11,28). To study eventual differences between centromere and telomere spatial organization, we developed a method that finds the radial distribution of multiple nuclear components in a single nucleus.…”

Section: Changes In Spatial Distribution During Activation Of Apoptosismentioning

confidence: 99%

Segmentation and analysis of the three‐dimensional redistribution of nuclear components in human mesenchymal stem cells

et al. 2008

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To better understand the impact of changes in nuclear architecture on nuclear functions, it is essential to quantitatively elucidate the three-dimensional organization of nuclear components using image processing tools. We have developed a novel image segmentation method, which involves a contrast enhancement and a subsequent thresholding step. In addition, we have developed a new segmentation method of the nuclear volume using the fluorescent background signal of a probe. After segmentation of the nucleus, a first-order normalization is performed on the signal positions of the component of interest to correct for the shape of the nucleus. This method allowed us to compare various signal positions within a single nucleus, and also on pooled data obtained from multiple nuclei, which may vary in size and shape. The algorithms have been tested by analyzing the spatial localization of nuclear bodies in relation to the nuclear center. Next, we used this new tool to study the change in the spatial distribution of nuclear components in cells before and after caspase-8 activation, which leads to cell death. Compared to the morphological TopHat method, this method gives similar but significantly faster results. A clear shift in the radial distribution of centromeres has been found, while the radial distribution of telomeres was changed much less. In addition, we have used this new tool to follow changes in the spatial distribution of two nuclear components in the same nucleus during activation of apoptosis. We show that after caspase-8 activation, when centromeres shift to a peripheral localization, the spatial distribution of PML-NBs does not change while that of centromeres did. We propose that the use of this new image segmentation method will contribute to a better understanding of the 3D spatial organization of the cell nucleus. ' 2008 International Society for Advancement of CytometryKey terms confocal microscopy; image processing; caspase-8; apoptosis; telomeres; centromeres THE major function of the cell nucleus is to regulate gene activity, which depends on well-studied molecular mechanisms such as transcription, pre-mRNA splicing, and ribosome assembly. In contrast to what is known about the molecular regulation of these mechanisms, far less is understood about the extent that the dynamics of nuclear components and the three-dimensional (3D) structural organization of the nucleus contribute to the regulation of nuclear functions. Recent models of high-order genome organization suggest a nonrandom spatial localization of chromosome territories in the interphase nucleus (1). Also subchromosomal domains are suggested to be nonrandomly positioned. Centromeres containing pericentric satellite repeats show a preferential peripheral orientation in G0-arrested human cells (2) as well as in differentiated cells (2-4). Telomeres, which are satellite repeats at the ends of chromosomes, reveal a cell cycle-dependent localization in B-lymphocytes. Throughout the cell cycle, telomeres exhibit a spherical organizatio...

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“…The adenovirus engineered with the H19 enhancer / DMD-H19 promoter complex induces apoptosis only in the cancer cells with loss of imprinting of the insulin-like growth factor 2 gene (IGF2). Artificial "death switches" are introduced into cancer cells by adenoviruses to initiate apoptosis [124][125][126][127][128][129][130]. Replication-competent adenovirusmediated suicide gene therapy (ReCAP) is in the clinical trials for newly-diagnosed prostate cancer.…”

Section: Vectors Of Suicidal Genesmentioning

confidence: 99%

“…the human, constitutively active caspases expressed from the viral vectors, effectively induce apoptosis of the human embryonic kidney and breast cancer cell lines [128]. Transgenic expression of 'death switches', bax and caspase 9, triggers apoptotic cascades and kills the cancer cells [126][127][128][129]. The final stage of many different routes leading to cancer cells' suicides is executed by DNases, what is manifested by hallmarks of apoptosis: collapse of chromatin and disintegration of the genomic DNA.…”

Section: Mechanisms Of Inducing Cancer Cells' Deathmentioning

confidence: 99%

Frontiers in Suicide Gene Therapy of Cancer

Malecki¹

2012

J Genet Syndr Gene Ther

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The National Cancer Institute (NCI) and the American Cancer Society (ACS) predict that 1,638,910 men and women will be diagnosed with cancer in the USA in 2012. Nearly 577,190 patients will die of cancer of all sites this year. Patients undergoing current systemic therapies will suffer multiple side effects from nausea to infertility. Potential parents, when diagnosed with cancer, will have to deposit oocytes and sperms prior to starting systemic radiation or chemo-therapy for the future genetic testing and in vitro fertilization, while trying to avoid risks of iatrogenic mutations in their germ cells. Otherwise, children of parents treated with systemic therapies, will be at high risk of developing genetic disorders. According to these predictions, this year will carry another, very poor therapeutic record again.The ultimate goal of cancer therapy is the complete elimination of all cancer cells, while leaving all healthy cells unharmed. One of the most promising therapeutic strategies in this regard is cancer suicide gene therapy (CSGT), which is rapidly progressing into new frontiers.The therapeutic success, in CSGT, is primarily contingent upon precision in delivery of the therapeutic transgenes to the cancer cells only. This is addressed by discovering and targeting unique or / and over-expressed biomarkers displayed on the cancer cells and cancer stem cells. Specificity of cancer therapeutic effects is further enhanced by designing the DNA constructs, which put the therapeutic genes under the control of the cancer cell specific promoters. The delivery of the suicidal genes to the cancer cells involves viral, as well as synthetic vectors, which are guided by cancer specific antibodies and ligands. The delivery options also include engineered stem cells with tropisms towards cancers. Main mechanisms inducing the cancer cells' deaths include: transgenic expression of thymidine kinases, cytosine deaminases, intracellular antibodies, telomeraseses, caspases, DNases. Precautions are undertaken to eliminate the risks associated with transgenesis.Progress in genomics and proteomics should help us in identifying the cancer specific biomarkers and metabolic pathways for developing new strategies towards clinical trials of targeted and personalized gene therapy of cancer.

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Development of an inducible suicide gene system based on human caspase 8

Cited by 39 publications

References 52 publications

Characterization of an immuno ‘stealth’ derivative of the herpes simplex virus thymidine-kinase gene

Characterization of an immuno ‘stealth’ derivative of the herpes simplex virus thymidine-kinase gene

Segmentation and analysis of the three‐dimensional redistribution of nuclear components in human mesenchymal stem cells

Frontiers in Suicide Gene Therapy of Cancer

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