Plant thymidine kinase 1

a novel efficient suicide gene for malignant glioma therapy

Zahidul Khan, Wolfgang Knecht,, Mette Willer, , Elzbieta Rozpedowska, Peter Kristoffersen, Anders Ranegaard Clausen,, Birgitte Munch-Petersen, Per M. Almqvist, , Zoran Gojkovic, Piskur ,Jure, Tomas J. Ekstrom,

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    The prognosis for malignant gliomas remains poor, and new treatments are urgently needed. Targeted suicide gene therapy exploits the enzymatic conversion of a prodrug, such as a nucleoside analog, into a cytotoxic compound. Although this therapeutic strategy has been considered a promising regimen for central nervous system (CNS) tumors, several obstacles have been encountered such as inefficient gene transfer to the tumor cells, limited prodrug penetration into the CNS, and inefficient enzymatic activity of the suicide gene. We report here the cloning and successful application of a novel thymidine kinase 1 (TK1) from the tomato plant, with favorable characteristics in vitro and in vivo. This enzyme (toTK1) is highly specific for the nucleoside analog prodrug zidovudine (azidothymidine, AZT), which is known to penetrate the blood-brain barrier. An important feature of toTK1 is that it efficiently phosphorylates its substrate AZT not only to AZT monophosphate, but also to AZT diphosphate, with excellent kinetics. The efficiency of the toTK1/AZT system was confirmed when toTK1-transduced human glioblastoma (GBM) cells displayed a 500-fold increased sensitivity to AZT compared with wild-type cells. In addition, when neural progenitor cells were used as delivery vectors for toTK1 in intracranial GBM xenografts in nude rats, substantial attenuation of tumor growth was achieved in animals exposed to AZT, and survival of the animals was significantly improved compared with controls. The novel toTK1/AZT suicide gene therapy system in combination with stem cell mediated gene delivery promises new treatment of malignant gliomas.
    Original languageEnglish
    JournalNeuro-Oncology
    Volume12
    Issue number6
    Pages (from-to)549-558
    Number of pages9
    ISSN1522-8517
    DOIs
    Publication statusPublished - 2010

    Cite this

    Khan, Z., Knecht, W., Willer, , M., Rozpedowska, E., Kristoffersen, P., Clausen, A. R., ... Ekstrom, , T. J. (2010). Plant thymidine kinase 1: a novel efficient suicide gene for malignant glioma therapy. Neuro-Oncology, 12(6), 549-558. https://doi.org/10.1093/neuonc/nop067
    Khan, Zahidul ; Knecht, Wolfgang ; Willer, , Mette ; Rozpedowska, Elzbieta ; Kristoffersen, Peter ; Clausen, Anders Ranegaard ; Munch-Petersen, Birgitte ; Almqvist, , Per M. ; Gojkovic, Zoran ; ,Jure, Piskur ; Ekstrom, , Tomas J. / Plant thymidine kinase 1 : a novel efficient suicide gene for malignant glioma therapy. In: Neuro-Oncology. 2010 ; Vol. 12, No. 6. pp. 549-558.
    @article{5af19be6deb041d1988bb1a4fc049847,
    title = "Plant thymidine kinase 1: a novel efficient suicide gene for malignant glioma therapy",
    abstract = "The prognosis for malignant gliomas remains poor, and new treatments are urgently needed. Targeted suicide gene therapy exploits the enzymatic conversion of a prodrug, such as a nucleoside analog, into a cytotoxic compound. Although this therapeutic strategy has been considered a promising regimen for central nervous system (CNS) tumors, several obstacles have been encountered such as inefficient gene transfer to the tumor cells, limited prodrug penetration into the CNS, and inefficient enzymatic activity of the suicide gene. We report here the cloning and successful application of a novel thymidine kinase 1 (TK1) from the tomato plant, with favorable characteristics in vitro and in vivo. This enzyme (toTK1) is highly specific for the nucleoside analog prodrug zidovudine (azidothymidine, AZT), which is known to penetrate the blood-brain barrier. An important feature of toTK1 is that it efficiently phosphorylates its substrate AZT not only to AZT monophosphate, but also to AZT diphosphate, with excellent kinetics. The efficiency of the toTK1/AZT system was confirmed when toTK1-transduced human glioblastoma (GBM) cells displayed a 500-fold increased sensitivity to AZT compared with wild-type cells. In addition, when neural progenitor cells were used as delivery vectors for toTK1 in intracranial GBM xenografts in nude rats, substantial attenuation of tumor growth was achieved in animals exposed to AZT, and survival of the animals was significantly improved compared with controls. The novel toTK1/AZT suicide gene therapy system in combination with stem cell mediated gene delivery promises new treatment of malignant gliomas.",
    keywords = "EXPERIMENTAL BRAIN-TUMORS, CEREBROSPINAL-FLUID, IN-VITRO, CELLS, GANCICLOVIR, DELIVERY, MUTANTS, PHARMACOKINETICS; , COMMUNICATION, PROGENITORS",
    author = "Zahidul Khan and Wolfgang Knecht, and Mette Willer, and Elzbieta Rozpedowska and Peter Kristoffersen and Clausen,, {Anders Ranegaard} and Birgitte Munch-Petersen and Almqvist,, {Per M.} and Zoran Gojkovic and Piskur ,Jure and Ekstrom,, {Tomas J.}",
    year = "2010",
    doi = "10.1093/neuonc/nop067",
    language = "English",
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    Khan, Z, Knecht, W, Willer, , M, Rozpedowska, E, Kristoffersen, P, Clausen, AR, Munch-Petersen, B, Almqvist, , PM, Gojkovic, Z, ,Jure, P & Ekstrom, , TJ 2010, 'Plant thymidine kinase 1: a novel efficient suicide gene for malignant glioma therapy', Neuro-Oncology, vol. 12, no. 6, pp. 549-558. https://doi.org/10.1093/neuonc/nop067

    Plant thymidine kinase 1 : a novel efficient suicide gene for malignant glioma therapy. / Khan, Zahidul; Knecht, Wolfgang; Willer, , Mette; Rozpedowska, Elzbieta; Kristoffersen, Peter; Clausen, Anders Ranegaard; Munch-Petersen, Birgitte; Almqvist, , Per M.; Gojkovic, Zoran; ,Jure, Piskur; Ekstrom, , Tomas J.

    In: Neuro-Oncology, Vol. 12, No. 6, 2010, p. 549-558.

    Research output: Contribution to journalJournal articleResearchpeer-review

    TY - JOUR

    T1 - Plant thymidine kinase 1

    T2 - a novel efficient suicide gene for malignant glioma therapy

    AU - Khan, Zahidul

    AU - Knecht,, Wolfgang

    AU - Willer, , Mette

    AU - Rozpedowska, Elzbieta

    AU - Kristoffersen, Peter

    AU - Clausen,, Anders Ranegaard

    AU - Munch-Petersen, Birgitte

    AU - Almqvist, , Per M.

    AU - Gojkovic, Zoran

    AU - ,Jure, Piskur

    AU - Ekstrom, , Tomas J.

    PY - 2010

    Y1 - 2010

    N2 - The prognosis for malignant gliomas remains poor, and new treatments are urgently needed. Targeted suicide gene therapy exploits the enzymatic conversion of a prodrug, such as a nucleoside analog, into a cytotoxic compound. Although this therapeutic strategy has been considered a promising regimen for central nervous system (CNS) tumors, several obstacles have been encountered such as inefficient gene transfer to the tumor cells, limited prodrug penetration into the CNS, and inefficient enzymatic activity of the suicide gene. We report here the cloning and successful application of a novel thymidine kinase 1 (TK1) from the tomato plant, with favorable characteristics in vitro and in vivo. This enzyme (toTK1) is highly specific for the nucleoside analog prodrug zidovudine (azidothymidine, AZT), which is known to penetrate the blood-brain barrier. An important feature of toTK1 is that it efficiently phosphorylates its substrate AZT not only to AZT monophosphate, but also to AZT diphosphate, with excellent kinetics. The efficiency of the toTK1/AZT system was confirmed when toTK1-transduced human glioblastoma (GBM) cells displayed a 500-fold increased sensitivity to AZT compared with wild-type cells. In addition, when neural progenitor cells were used as delivery vectors for toTK1 in intracranial GBM xenografts in nude rats, substantial attenuation of tumor growth was achieved in animals exposed to AZT, and survival of the animals was significantly improved compared with controls. The novel toTK1/AZT suicide gene therapy system in combination with stem cell mediated gene delivery promises new treatment of malignant gliomas.

    AB - The prognosis for malignant gliomas remains poor, and new treatments are urgently needed. Targeted suicide gene therapy exploits the enzymatic conversion of a prodrug, such as a nucleoside analog, into a cytotoxic compound. Although this therapeutic strategy has been considered a promising regimen for central nervous system (CNS) tumors, several obstacles have been encountered such as inefficient gene transfer to the tumor cells, limited prodrug penetration into the CNS, and inefficient enzymatic activity of the suicide gene. We report here the cloning and successful application of a novel thymidine kinase 1 (TK1) from the tomato plant, with favorable characteristics in vitro and in vivo. This enzyme (toTK1) is highly specific for the nucleoside analog prodrug zidovudine (azidothymidine, AZT), which is known to penetrate the blood-brain barrier. An important feature of toTK1 is that it efficiently phosphorylates its substrate AZT not only to AZT monophosphate, but also to AZT diphosphate, with excellent kinetics. The efficiency of the toTK1/AZT system was confirmed when toTK1-transduced human glioblastoma (GBM) cells displayed a 500-fold increased sensitivity to AZT compared with wild-type cells. In addition, when neural progenitor cells were used as delivery vectors for toTK1 in intracranial GBM xenografts in nude rats, substantial attenuation of tumor growth was achieved in animals exposed to AZT, and survival of the animals was significantly improved compared with controls. The novel toTK1/AZT suicide gene therapy system in combination with stem cell mediated gene delivery promises new treatment of malignant gliomas.

    KW - EXPERIMENTAL BRAIN-TUMORS

    KW - CEREBROSPINAL-FLUID

    KW - IN-VITRO

    KW - CELLS

    KW - GANCICLOVIR

    KW - DELIVERY

    KW - MUTANTS

    KW - PHARMACOKINETICS;

    KW - COMMUNICATION

    KW - PROGENITORS

    U2 - 10.1093/neuonc/nop067

    DO - 10.1093/neuonc/nop067

    M3 - Journal article

    VL - 12

    SP - 549

    EP - 558

    JO - Neuro-Oncology

    JF - Neuro-Oncology

    SN - 1522-8517

    IS - 6

    ER -

    Khan Z, Knecht, W, Willer, M, Rozpedowska E, Kristoffersen P, Clausen, AR et al. Plant thymidine kinase 1: a novel efficient suicide gene for malignant glioma therapy. Neuro-Oncology. 2010;12(6):549-558. https://doi.org/10.1093/neuonc/nop067