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Targeting JNK for therapeutic depletion of stem-like glioblastoma cells.

Matsuda K, Sato A, Okada M, Shibuya K, Seino S, Suzuki K, Watanabe E, Narita Y, Shibui S, Kayama T, Kitanaka C - Sci Rep (2012)

Bottom Line: Control of the stem-like tumour cell population is considered key to realizing the long-term survival of patients with glioblastoma, one of the most devastating human malignancies.To date, possible therapeutic targets and targeting methods have been described, but none has yet proven to target stem-like glioblastoma cells in the brain to the extent necessary to provide a survival benefit.Here we show that targeting JNK in vivo, the activity of which is required for the maintenance of stem-like glioblastoma cells, via transient, systemic administration of a small-molecule JNK inhibitor depletes the self-renewing and tumour-initiating populations within established tumours, inhibits tumour formation by stem-like glioblastoma cells in the brain, and provide substantial survival benefit without evidence of adverse events.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Cancer Science, Yamagata University School of Medicine, Yamagata, 990-9585, Japan.

ABSTRACT
Control of the stem-like tumour cell population is considered key to realizing the long-term survival of patients with glioblastoma, one of the most devastating human malignancies. To date, possible therapeutic targets and targeting methods have been described, but none has yet proven to target stem-like glioblastoma cells in the brain to the extent necessary to provide a survival benefit. Here we show that targeting JNK in vivo, the activity of which is required for the maintenance of stem-like glioblastoma cells, via transient, systemic administration of a small-molecule JNK inhibitor depletes the self-renewing and tumour-initiating populations within established tumours, inhibits tumour formation by stem-like glioblastoma cells in the brain, and provide substantial survival benefit without evidence of adverse events. Our findings not only implicate JNK in the maintenance of stem-like glioblastoma cells but also demonstrate that JNK is a viable, clinically relevant therapeutic target in the control of stem-like glioblastoma cells.

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Related in: MedlinePlus

Transient in vivo JNK inhibition deprives stem-like glioblastoma cells of tumour-initiating potential and depletes established tumours of self-renewing and tumour-initiating glioblastoma cell populations.(a) Mice implanted subcutaneously with stem-like glioblastoma cells (TGS01, 1×106 cells) were administered intraperitoneal injection of the control vehicle (DMSO) or SP600125 (40 mg/kg/day), for 5 days starting on the next day of implantation (3 mice per group). Left, tumour volume at the indicated time points (mean ± s.d.). Right, representative images of mice 28 days after implantation. *P < 0.05. (b–d) Mice subcutaneously implanted with stem-like glioblastoma cells (TGS01, 1×106 cells) were administered, after tumour formation (tumour volume range, 224–364 mm3), intraperitoneal injection of the control vehicle (DMSO) or SP600125 (40 mg/kg/day) for 5 days (3 mice per group). On the next day of the final drug treatment, mice were sacrificed and dissociated tumour cells were subjected to serial sphere formation assays (b). Left, photomicrographs of the spheres (scale bars, 200 μm). Right, number of spheres formed (mean ± s.d. of triplicate cultures derived from a single tumour of each treatment group). Essentially identical results were obtained from analysis of the remaining 2 tumours of each group. Alternatively, the dissected tumour cells (1×106 cells per mouse) were transplanted subcutaneously into the right flank (5 mice per group) (c). Left, tumour volume at the indicated time points (mean ± s.d.). Right, at the end of the observation period (28 days after transplantation), mice were sacrificed and the weight of the secondary tumours was measured. Middle, representative image of the secondary tumours derived from the SP600125- and control-treated primary tumours. Serial dilutions of the dissected tumour cells derived from the SP600125- and control-treated primary tumours were also transplanted intracranially (d). Right, survival as evaluated by Kaplan-Meier analysis (3 mice per group). Left, representative images of haematoxylin and eosin staining of the brain sections of mice that had undergone transplantation of cells (1×104) from primary tumours treated with SP600125 or the control vehicle (DMSO) and were sacrificed at 20 days after transplantation. *P < 0.05.
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f4: Transient in vivo JNK inhibition deprives stem-like glioblastoma cells of tumour-initiating potential and depletes established tumours of self-renewing and tumour-initiating glioblastoma cell populations.(a) Mice implanted subcutaneously with stem-like glioblastoma cells (TGS01, 1×106 cells) were administered intraperitoneal injection of the control vehicle (DMSO) or SP600125 (40 mg/kg/day), for 5 days starting on the next day of implantation (3 mice per group). Left, tumour volume at the indicated time points (mean ± s.d.). Right, representative images of mice 28 days after implantation. *P < 0.05. (b–d) Mice subcutaneously implanted with stem-like glioblastoma cells (TGS01, 1×106 cells) were administered, after tumour formation (tumour volume range, 224–364 mm3), intraperitoneal injection of the control vehicle (DMSO) or SP600125 (40 mg/kg/day) for 5 days (3 mice per group). On the next day of the final drug treatment, mice were sacrificed and dissociated tumour cells were subjected to serial sphere formation assays (b). Left, photomicrographs of the spheres (scale bars, 200 μm). Right, number of spheres formed (mean ± s.d. of triplicate cultures derived from a single tumour of each treatment group). Essentially identical results were obtained from analysis of the remaining 2 tumours of each group. Alternatively, the dissected tumour cells (1×106 cells per mouse) were transplanted subcutaneously into the right flank (5 mice per group) (c). Left, tumour volume at the indicated time points (mean ± s.d.). Right, at the end of the observation period (28 days after transplantation), mice were sacrificed and the weight of the secondary tumours was measured. Middle, representative image of the secondary tumours derived from the SP600125- and control-treated primary tumours. Serial dilutions of the dissected tumour cells derived from the SP600125- and control-treated primary tumours were also transplanted intracranially (d). Right, survival as evaluated by Kaplan-Meier analysis (3 mice per group). Left, representative images of haematoxylin and eosin staining of the brain sections of mice that had undergone transplantation of cells (1×104) from primary tumours treated with SP600125 or the control vehicle (DMSO) and were sacrificed at 20 days after transplantation. *P < 0.05.

Mentions: Having established the essential role of JNK in the maintenance of the tumour-initiating potential of stem-like glioblastoma cells, we next sought to determine if JNK could be an in vivo target in controlling the tumour-initiating potential of glioblastoma cells. To this end, we tested the effect of systemic administration of SP600125 on tumour formation by stem-like glioblastoma cells. We started in this study from a much less intense, short-term regimen (intraperitoneal administration of SP600125 at 40 mg/kg/day for 5 consecutive days) compared to the regimen used in a previous study (daily intraperitoneal administration of SP600125 at 50 mg/kg/day for 1 month to nude mice)20, and evaluated the effectiveness of the regimen against subcutaneous tumour formation to see if intensification of the treatment schedule is needed. Rather unexpectedly, even with this starting, less intense regimen of drug administration, we observed a significant inhibitory effect of SP600125 treatment compared to the control treatment against tumour formation either by stem-like glioblastoma cells directly derived from a patient (TGS01; Fig. 4a) or by stem-like U87GS cells derived from the conventional, serum-cultured cell line U87 (Supplementary Fig. 9d).


Targeting JNK for therapeutic depletion of stem-like glioblastoma cells.

Matsuda K, Sato A, Okada M, Shibuya K, Seino S, Suzuki K, Watanabe E, Narita Y, Shibui S, Kayama T, Kitanaka C - Sci Rep (2012)

Transient in vivo JNK inhibition deprives stem-like glioblastoma cells of tumour-initiating potential and depletes established tumours of self-renewing and tumour-initiating glioblastoma cell populations.(a) Mice implanted subcutaneously with stem-like glioblastoma cells (TGS01, 1×106 cells) were administered intraperitoneal injection of the control vehicle (DMSO) or SP600125 (40 mg/kg/day), for 5 days starting on the next day of implantation (3 mice per group). Left, tumour volume at the indicated time points (mean ± s.d.). Right, representative images of mice 28 days after implantation. *P < 0.05. (b–d) Mice subcutaneously implanted with stem-like glioblastoma cells (TGS01, 1×106 cells) were administered, after tumour formation (tumour volume range, 224–364 mm3), intraperitoneal injection of the control vehicle (DMSO) or SP600125 (40 mg/kg/day) for 5 days (3 mice per group). On the next day of the final drug treatment, mice were sacrificed and dissociated tumour cells were subjected to serial sphere formation assays (b). Left, photomicrographs of the spheres (scale bars, 200 μm). Right, number of spheres formed (mean ± s.d. of triplicate cultures derived from a single tumour of each treatment group). Essentially identical results were obtained from analysis of the remaining 2 tumours of each group. Alternatively, the dissected tumour cells (1×106 cells per mouse) were transplanted subcutaneously into the right flank (5 mice per group) (c). Left, tumour volume at the indicated time points (mean ± s.d.). Right, at the end of the observation period (28 days after transplantation), mice were sacrificed and the weight of the secondary tumours was measured. Middle, representative image of the secondary tumours derived from the SP600125- and control-treated primary tumours. Serial dilutions of the dissected tumour cells derived from the SP600125- and control-treated primary tumours were also transplanted intracranially (d). Right, survival as evaluated by Kaplan-Meier analysis (3 mice per group). Left, representative images of haematoxylin and eosin staining of the brain sections of mice that had undergone transplantation of cells (1×104) from primary tumours treated with SP600125 or the control vehicle (DMSO) and were sacrificed at 20 days after transplantation. *P < 0.05.
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f4: Transient in vivo JNK inhibition deprives stem-like glioblastoma cells of tumour-initiating potential and depletes established tumours of self-renewing and tumour-initiating glioblastoma cell populations.(a) Mice implanted subcutaneously with stem-like glioblastoma cells (TGS01, 1×106 cells) were administered intraperitoneal injection of the control vehicle (DMSO) or SP600125 (40 mg/kg/day), for 5 days starting on the next day of implantation (3 mice per group). Left, tumour volume at the indicated time points (mean ± s.d.). Right, representative images of mice 28 days after implantation. *P < 0.05. (b–d) Mice subcutaneously implanted with stem-like glioblastoma cells (TGS01, 1×106 cells) were administered, after tumour formation (tumour volume range, 224–364 mm3), intraperitoneal injection of the control vehicle (DMSO) or SP600125 (40 mg/kg/day) for 5 days (3 mice per group). On the next day of the final drug treatment, mice were sacrificed and dissociated tumour cells were subjected to serial sphere formation assays (b). Left, photomicrographs of the spheres (scale bars, 200 μm). Right, number of spheres formed (mean ± s.d. of triplicate cultures derived from a single tumour of each treatment group). Essentially identical results were obtained from analysis of the remaining 2 tumours of each group. Alternatively, the dissected tumour cells (1×106 cells per mouse) were transplanted subcutaneously into the right flank (5 mice per group) (c). Left, tumour volume at the indicated time points (mean ± s.d.). Right, at the end of the observation period (28 days after transplantation), mice were sacrificed and the weight of the secondary tumours was measured. Middle, representative image of the secondary tumours derived from the SP600125- and control-treated primary tumours. Serial dilutions of the dissected tumour cells derived from the SP600125- and control-treated primary tumours were also transplanted intracranially (d). Right, survival as evaluated by Kaplan-Meier analysis (3 mice per group). Left, representative images of haematoxylin and eosin staining of the brain sections of mice that had undergone transplantation of cells (1×104) from primary tumours treated with SP600125 or the control vehicle (DMSO) and were sacrificed at 20 days after transplantation. *P < 0.05.
Mentions: Having established the essential role of JNK in the maintenance of the tumour-initiating potential of stem-like glioblastoma cells, we next sought to determine if JNK could be an in vivo target in controlling the tumour-initiating potential of glioblastoma cells. To this end, we tested the effect of systemic administration of SP600125 on tumour formation by stem-like glioblastoma cells. We started in this study from a much less intense, short-term regimen (intraperitoneal administration of SP600125 at 40 mg/kg/day for 5 consecutive days) compared to the regimen used in a previous study (daily intraperitoneal administration of SP600125 at 50 mg/kg/day for 1 month to nude mice)20, and evaluated the effectiveness of the regimen against subcutaneous tumour formation to see if intensification of the treatment schedule is needed. Rather unexpectedly, even with this starting, less intense regimen of drug administration, we observed a significant inhibitory effect of SP600125 treatment compared to the control treatment against tumour formation either by stem-like glioblastoma cells directly derived from a patient (TGS01; Fig. 4a) or by stem-like U87GS cells derived from the conventional, serum-cultured cell line U87 (Supplementary Fig. 9d).

Bottom Line: Control of the stem-like tumour cell population is considered key to realizing the long-term survival of patients with glioblastoma, one of the most devastating human malignancies.To date, possible therapeutic targets and targeting methods have been described, but none has yet proven to target stem-like glioblastoma cells in the brain to the extent necessary to provide a survival benefit.Here we show that targeting JNK in vivo, the activity of which is required for the maintenance of stem-like glioblastoma cells, via transient, systemic administration of a small-molecule JNK inhibitor depletes the self-renewing and tumour-initiating populations within established tumours, inhibits tumour formation by stem-like glioblastoma cells in the brain, and provide substantial survival benefit without evidence of adverse events.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Cancer Science, Yamagata University School of Medicine, Yamagata, 990-9585, Japan.

ABSTRACT
Control of the stem-like tumour cell population is considered key to realizing the long-term survival of patients with glioblastoma, one of the most devastating human malignancies. To date, possible therapeutic targets and targeting methods have been described, but none has yet proven to target stem-like glioblastoma cells in the brain to the extent necessary to provide a survival benefit. Here we show that targeting JNK in vivo, the activity of which is required for the maintenance of stem-like glioblastoma cells, via transient, systemic administration of a small-molecule JNK inhibitor depletes the self-renewing and tumour-initiating populations within established tumours, inhibits tumour formation by stem-like glioblastoma cells in the brain, and provide substantial survival benefit without evidence of adverse events. Our findings not only implicate JNK in the maintenance of stem-like glioblastoma cells but also demonstrate that JNK is a viable, clinically relevant therapeutic target in the control of stem-like glioblastoma cells.

Show MeSH
Related in: MedlinePlus