| 研究課題/領域番号 |
18K16553
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| 研究機関 | 金沢大学 |
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研究代表者 |
Pyko Ilya 金沢大学, がん進展制御研究所, 博士研究員 (00731853)
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| 研究期間 (年度) |
2018-04-01 – 2020-03-31
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| キーワード | glioblastoma / temozolomide / GBM stem-like cells / GSK3β |
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| 研究実績の概要 |
Studies in vitro and in animal model were carried out complementary to optimize treatment of glioblastoma (GBM) by enhancing anti-tumor effect by combination of GSK3β inhibition and temozolomide (TMZ). For in vitro study, we examined effect of GSK3β inhibition and TMZ on patient-derived GBM stem-like cells (SCs). I found that GSK3β inhibition enhances effect of TMZ against GBM-SCs and participate in regulation of GBM stemness phenotype. For animal model study, I have developed software pharmacokinetics model and determined optimal concentration of GSK3β inhibitor for continuous intra-tumor infusion by subcutaneous pumps for treatment of orthotopic GBM models and at present we examine effects of continuous intra-tumor infusion of GSK3β inhibitor, against GBM in mice bearing human GBM-SCs. Our experiments showed that GSK3β inhibition is effective for treatment of experimental GBM generated by inoculation of most malignant GBM-SCs characterized by shortest survival in control animals. This observation encouraged us to investigate biological mechanisms by which GSK3β regulate GBM stemness phenotype by starting from the screening for changes in stem cell markers’ expression in GBM-SCs under GSK3β inhibition.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
The following steps have been performed for the study. (1) preparation of 30 GBM-SC cultures from patients with primary GBM; among which 25 with high O6-methylguanine DNA methyltransferase (MGMT) expression and resistant to temozolomide were used in experiments;
(2) I have investigated the effects of TMZ, GSK3β inhibition and primary GBM-SCs stemness phenotype by proliferation assay, and by evaluation of stem cell markers’ gene expression by QRT-PCR and protein expression by Western Blot;
(3) I have developed three orthotopic GBM models generated by using human GBM-SCs that adequately represent human GBM;
(4) I have developed software pharmacokinetics model and determined optimal concentration of GSK3β inhibitor for continuous intra-tumor infusion by subcutaneous pumps for treatment of orthotopic GBM models;
(5) I have investigated the effects of continuous intra-tumor infusion GSK3β inhibitor and on neurological state and survival of mice with orthotopic GBM models generated by using human GBM-SCs and confirmed high anti-tumor effect of the treatment in the model generated by inoculation of most malignant GBM-SCs characterized by shortest control animal survival.
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| 今後の研究の推進方策 |
Studies in vitro and in animal model will be carried out complementary to optimize treatment of glioblastoma (GBM) by enhancing anti-tumor effect by combination of GSK3β inhibition and temozolomide (TMZ).
For in vitro study, we examine effects of GSK3β inhibition, TMZ and their combination against GBM-SCs. To address our working hypothesis of regulation of stemness phenotype of GBM-SCs via GSK3β-mediated signaling, we investigate stem cell markers’ expression in GBM-SCs under GSK3β inhibition.
For animal model study, we examine effects and underlying mechanism of intracranial transplantation of MSCs, against GBM in mice bearing human GBM-SCs. We observe survival as well as consequence of neurological state in mice following transplantation of the respective cells.
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