scholarly journals Use of a herpes thymidine kinase/neomycin phosphotransferase chimeric gene for metabolic suicide gene transfer

2000 ◽  
Vol 7 (4) ◽  
pp. 574-580 ◽  
Author(s):  
Fabio Candotti ◽  
Riad Agbaria ◽  
Craig A Mullen ◽  
Renaud Touraine ◽  
Jan Balzarini ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Thymidine Kinase ◽  
Suicide Gene ◽  
Chimeric Gene ◽  
Neomycin Phosphotransferase

Improved Suicide Gene Therapy: Lentiviral Gene Transfer of Equine Herpes Virus Type 4 Thymidine Kinase into Target Cells.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 5251-5251
Author(s):  
Takeya Sato ◽  
Anton Neschadim ◽  
Vanessa I. Rasaiah ◽  
Manfred Konrad ◽  
Daniel H. Fowler ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Gene Transfer ◽  
Thymidine Kinase ◽  
Virus Type ◽  
Herpes Virus ◽  
Suicide Gene ◽  
Suicide Gene Therapy ◽  
Jurkat Cells ◽  
Target Cells ◽  
Herpès Virus

Abstract Herpes virus type 1 thymidine kinase (HSV1-TK) with ganciclovir (GCV) prodrug treatment is the most widely used approach for suicide gene therapy. This ‘suicide’ strategy allows direct reduction of tumors and clearance of donor cells should graft-versus-host disease (GvHD) arise after bone marrow transplantation. Given recent clinical outcomes, this suicide approach may also provide a key safety component for therapeutic gene transfer vectors that integrate. Although suicide gene therapy using HSV1-TK-encoding oncoretroviral vectors has been evaluated in the clinic, the success of this approach has been relatively modest. Reasons for this include: low gene transfer efficacy, reduced expression of the suicide gene, and insufficient conversion of substrate. Our goal is to overcome these limitations by using a novel lentiviral vector (LV) encoding an alternative kinase/prodrug combination. The rational for our innovative suicide gene therapy strategy is two-fold: 1) Lentiviral vectors can efficiently transduce not only dividing cells but also non-dividing cells. 2) Applying a faster viral enzyme like equine herpes virus type 4 thymidine kinase (EHV4-TK) could be advantageous as it has been shown to be kinetically superior to HSV1-TK at GCV phosphorylation. The aim of this study is to evaluate whether LV-mediated gene modification of target cells with EHV4-TK can lead to efficient killing following GCV treatment. We first constructed a LV expression system carrying the wild-type EHV4-TK cDNA with an IRES element followed by a truncated form of human CD19 (hCD19Δ). Human CD19 was chosen as a cell surface marker to allow functional titering of virus and for immuno-enrichment of transduced cells prior to infusion since it is not expressed in the T cell lineage. The truncated form lacks the intracellular domain and therefore does not signal. Use of an IRES element can abrogate some variegated expression seen with vectors having dual promoters. The LV was pseudotyped with VSV-g and concentrated by ultracentrifugation. After one infection, Jurkat cells (human T cell leukemia) showed a more than 80% functional and stable transduction efficiency (MOI = 10). Using hCD19Δ as a selective marker, transduced Jurkat cells were enriched to over 95% positive by immuno-affinity sorting. EHV4-TK-transduced Jurkat cells exhibited increased cell killing in response to GCV treatment (the apoptotic cell indexes with or without GCV were 69.4 ± 1.5 % and 18.8 ± 1.7 %, respectively; n=3). Highly efficient transduction (more than 60%) of primary human T cells was accomplished by a three time exposure to virus over 36 hours at MOI of 20. Next, we found that GCV efficiently killed transduced primary human T cells in a dose dependent manner. We are now comparing the efficiency of GCV conversion by HSV1-TK and EHV4-TK using LV-transduced cells that express the similar protein levels. We are also evaluating intracellular levels of GCV metabolites by HPLC. These results demonstrate that our novel suicide gene therapy strategy has significant potential for many clinical applications.

Are hepatomas a good target for suicide gene therapy? An experimental study in rats using retroviral-mediated transfer of thymidine kinase gene

Surgery ◽  
1998 ◽  
Vol 123 (1) ◽  
pp. 19-24 ◽  
Author(s):  
Héléna Nagy ◽  
Yves Panis ◽  
Monique Fabre ◽  
Hubert Perrin ◽  
David Klatzmann ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Experimental Study ◽  
Thymidine Kinase ◽  
Suicide Gene ◽  
Suicide Gene Therapy ◽  
Thymidine Kinase Gene ◽  
Kinase Gene ◽  
Good Target

Adenoviral-Mediated Herpes Simplex Virus Thymidine Kinase Gene Transfer

Pancreas ◽  
1997 ◽  
Vol 15 (1) ◽  
pp. 25-34 ◽  
Author(s):  
Andreas Block ◽  
Shu-Hsia Chen ◽  
Ken-Ichiro Kosai ◽  
Milton Finegold ◽  
Savio L. C. Woo
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Gene Transfer ◽  
Thymidine Kinase ◽  
Thymidine Kinase Gene ◽  
Kinase Gene ◽  
Simplex Virus

scholarly journals Adenoviral-Mediated Herpes Simplex Virus-Thymidine Kinase Gene Transfer in Vivo for Treatment of Experimental Human Melanoma

1996 ◽  
Vol 106 (6) ◽  
pp. 1163-1168 ◽  
Author(s):  
Bernd Bonnekoh ◽  
David A. Greenhalgh ◽  
Donnie S. Bundman ◽  
Ken-ichiro Kosai ◽  
Shu-Hsia Chen ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Gene Transfer ◽  
Thymidine Kinase ◽  
Human Melanoma ◽  
Thymidine Kinase Gene ◽  
Kinase Gene ◽  
Simplex Virus

scholarly journals The war against cancer: Suicide gene therapy

2016 ◽  
Vol 2 (3) ◽  
pp. 139
Author(s):  
Muzeyyen Izmirli ◽  
Dilara Sonmez ◽  
Bulent Gogebakan
Keyword(s):  
Gene Therapy ◽  
Cytochrome P450 ◽  
Side Effects ◽  
Cancer Cells ◽  
Thymidine Kinase ◽  
Viral Vectors ◽  
Suicide Gene ◽  
Suicide Gene Therapy ◽  
Kinase Gene ◽  
Cancer Society

<p>The National Cancer Institute and the American Cancer Society announced that 1.6 million new cancer cases are projected to occur in the USA in 2016. One of the most innovative approaches against cancer is suicide gene therapy, in which suicide-inducing transgenes are introduced into cancer cells. When cancer treatments target the total elimination of tumor cells, there will be no side effects for normal cells. Cancer tissues are targeted through various targeted transport methods, followed by tissue-specific enzymes converting a systemically suitable prodrug into an active drug in the tumor. Suicidal genes are delivered by transporters, such as viral and non-viral vectors, into cancer cells. Suicide gene therapeutic strategies currently pursued are herpes simplex virus thymidine kinase gene with prodrug ganciclovir, cytosine deaminase gene, carboxyl esterase/irinotecan, varicella zoster virus thymidine kinase/6-methoxypurine arabinonucleoside, nitroreductase Nfsb/5-(aziridin-1-yl)-2,4-dinitrobenzamide, carboxypeptidase G2/4-[(2-chloroethyl)(2- mesyloxyethyl)amino]benzoyl-L-glutamic acid, cytochrome p450-isofosfamide, and cytochrome p450-cyclophosphamide. The goal of this review is to summarize the different suicide gene systems and gene delivery vectors addressed to cancer cells, with a particular emphasis on recently developed systems. Finally, we briefly describe the advantageous clinical applications and potential side effects of suicide gene therapy. </p>

scholarly journals A truncated herpes simplex virus thymidine kinase phosphorylates thymidine and nucleoside analogs and does not cause sterility in transgenic mice.

1995 ◽  
Vol 15 (10) ◽  
pp. 5322-5328 ◽  
Author(s):  
B Salomon ◽  
S Maury ◽  
L Loubière ◽  
M Caruso ◽  
R Onclercq ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Transgenic Mice ◽  
Thymidine Kinase ◽  
Nucleoside Analogs ◽  
Suicide Gene ◽  
Physiological Importance ◽  
Simplex Virus ◽  
High Level

Dividing eukaryotic cells expressing the herpes simplex virus type 1 thymidine kinase (TK) gene are sensitive to the cytotoxic effect of nucleoside analogs such as acyclovir or ganciclovir (GCV). Transgenic mice with cell-targeted expression of this conditional toxin have been used to create animals with temporally controlled cell-specific ablation. In these animal models, which allow the study of the physiological importance of a cell type, males are sterile. In this study, we showed that this phenomenon is due to testis-specific high-level expression of short TK transcripts initiated mainly upstream of the second internal ATG of the TK gene. This expression is DNA methylation independent. To obtain a suicide gene that does not cause male infertility, we generated and analyzed the properties of a truncated TK (delta TK) lacking the sequences upstream of the second ATG. We showed that when expressed at sufficient levels, the functional properties of delta TK are similar to those of TK in terms of thymidine or GCV phosphorylation. This translated into a similar GCV-dependent toxicity for delta TK- or TK-expressing cells, both in vitro and in transgenic mice. However, delta TK behaved differently from TK in two ways. First, it did not cause sterility in delta TK transgenic males. Second, low-level delta TK RNA expression did not confer sensitivity to GCV. The uses of delta TK in cell-specific ablation in transgenic mice and in gene therapy are discussed.

scholarly journals An Adenovirus Vector Containing the Suicide Gene Thymidine Kinase for a Broad Application in Cancer Gene Therapy

2002 ◽  
Vol 97 (4) ◽  
pp. 547-552 ◽  
Author(s):  
GS Magalhães ◽  
AR Muotri ◽  
MCN Marchetto ◽  
CFM Menck ◽  
AM Ventura
Keyword(s):  
Gene Therapy ◽  
Thymidine Kinase ◽  
Adenovirus Vector ◽  
Suicide Gene ◽  
Cancer Gene Therapy ◽  
Cancer Gene ◽  
Broad Application

Clinical-grade preparation of human natural regulatory T-cells encoding the thymidine kinase suicide gene as a safety gene

2008 ◽  
Vol 10 (8) ◽  
pp. 834-846 ◽  
Author(s):  
Maude Guillot-Delost ◽  
Mustapha Cheraï ◽  
Yamina Hamel ◽  
Michelle Rosenzwajg ◽  
Claude Baillou ◽  
...  
Keyword(s):  
T Cells ◽  
Regulatory T Cells ◽  
Thymidine Kinase ◽  
Suicide Gene ◽  
Clinical Grade
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