Grant-in-Aid for Scientific Research (B)
|Research Institution||Kobe University|
MISHIMA Yutaka Kobe Univ., School of Medicine, Honorary Professor, 医学部, 名誉教授 (10073743)
FUKUMORI Yoshinobu Faculty of Pharmaceutical Science, Kobe Gakuin Univ., Professor, 薬学部, 教授 (60102927)
YOSHINO Kazuo Shinshu Univ., Faculty of Science, Assistant Professor, 理学部, 助教授 (70143964)
SHIBAHARA Shigeki Tohoku Univ., School of Medicine, Professor, 医学部, 教授 (70206142)
新津 洋司郎 札幌医科大学, 医学部, 教授 (10045502)
TAMAKI Norihiko Kobe Univ., School of Medicine, Professor, 医学部, 教授 (10030941)
ONO Koji Research Reactor Institute, Kyoto Univ., Professor, 原子炉実験所, 教授 (90122407)
|Project Fiscal Year
1997 – 1998
Completed(Fiscal Year 1998)
|Budget Amount *help
¥12,700,000 (Direct Cost : ¥12,700,000)
Fiscal Year 1998 : ¥5,900,000 (Direct Cost : ¥5,900,000)
Fiscal Year 1997 : ¥6,800,000 (Direct Cost : ¥6,800,000)
|Keywords||cancer boron neutron capture therapy / in vivo gene-transfer / malignant melanoma / BPA / transferrin-conjugate gene vector / electroporation / isoform of MITF / melanin monomer complex / がん熱中性子捕捉療法 / in vivo遺伝子導入 / 悪性黒色腫 / トランスフェリン複合体ベクター / エレクトロポレーション / MITFアイソフォーム / BPAメラニンモノマー錯体 / キトサンナノパーティクル / 遺伝子導入|
During the term of project, April 1997 - March 1999 following research results have been obtained.
(1) Enhancement of therapeutic effect for the melanoma BNCT : Transfer of melanogenic gene, TRP-2 into less melanotic (pheomelanotic) human melanoma cells has enhanced their uptake of ^<10>B-BPA (Boronophenylalanine). Further, transfer of tyrosilase gene into fibroblast increased their uptake of ^<10>B-BPA.
(2) Isolation and identification of novel-isoform for MITF gene which can regulate the expression of melanogenic genes. Identification of SRF which may regulate the expression of TRP-2.
(3) Concerning mechanism of selective ^<10>B-BPA uptake into melanoma, we have found the formation of chemical complex between ^<10>B-BPA and melanin monomer/dimer, releasing lOB-boric acid which can be integrated melanin polymer formation.
II.Development of devices and methods for gene and ^<10>B-BPA delivery.
(1) Establishment of methodology to form transferrin-conjugate gene vector.
ulation method for nano-particle device as ^<10>B-BPA delivery carriers has proven its enhancing therapeutic effects in vovo.
(3) Electroporation method has enhanced uptake of ^<10>B-compounds by target cells in vitro and in vivo.
III.Therapeutic experiment using cancer-bearing animals.
(1) After the tyrosinase-transfer into amelanotic melanoma cells in culture with subsequent subcutaneous transplantation of these cells into mouse, we have found the markedly enhanced killing effect of ^<10>B-BPA BNCT experiment.
(2) After the in vivo tyrosinase gene-transfer into amelanotic melanoma proliferating in hamster skin by intra-tumor injection, we have found enhanced killing effect of ^<10>B-BPA BNCT in preliminary experiment.
(3) Using i.v. injection of transfenin-conjugate of HSV-tk gene to tumor bearing mice, we have found its dstinct enhanced therapeutic effect prolonging their survival period.
(4) After establishment of experimental model for amelanotic melanoma cells proliferating in brain, we have found increased ^<10>B-BPA uptake by these amelanotic melanoma cells, which were previously transfected with tyrosinase-gene, using ^<18>F-PET. Less