Development of New Strategy of Boron Neutron Capture Therapy to Cancer Combined with Tumor Specific Gene Delivery

结合肿瘤特异性基因递送的硼中子捕获治疗癌症新策略的开发

• 批准号: 11557092
• 负责人: YANAGIE Hironobu
• 金额: $ 8.58万
• 依托单位: TEIKYO UNIVERSITY (2001)The University of Tokyo (1999-2000)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11557092/
• 关键词: Boron Neutron Capture Therapv(BNCT); Drug Delivery System; Stealth Liposome; Qplex; Apoptosis; Boron Compound; Pulsed neutron generator; C型肝炎ウイルス細胞障害領域(INES領域); バルス式中性子発生装置; ドラッグデリバリーシステム; ステルスリポソーム; C型肝炎ウイルス細胞障害領域

We performed the study of development of enhanced effects of BNCT with tumor specific gene therapy.(1) We prepare a polyethylene-glycol (PEG) binding liposome entrapped ^<10>B compound for the delivery system. After thermal neutron irradiation of mice injected with ^<10>B-bare liposome or ^<10>B-PEG-liposome, AsPC-1 tumour growth was suppressed relative to controls. Colon 26 tumor was disappeared by BNCT with the injection of Transferrin-^<10>B-PEG-Liposome. Injection of Transferrin-^<10>B-PEG-Liposome and ^<10>B-PEG-liposome caused the greatest tumour suppression with thermal neutron irradiation in vivo.(2) We performed neutron capture autoradiography(NCAR) using CR-39 plastic track detectors. The CR-39 detectors attached with samples were exposed to thermal neutrons or cold neutrons at Rikkyo University, PSI, Institute of Jozef Stefan or Sakley Institute. We can increase the accumulation of ^<10>B atoms in the tumor tissues by PEG-liposome, which increase the retension in the blood f … More low and escape the phagocytosis by RES.(3) We developed the novel small sized palsed neutron generator. This neutron generator will be applied to BNCT.(4) It is necessary to increase the transfectional efficiency when we use non-viral vector on cancer gene therapy. We have prepared new typed plasmid DNA-cationic lipoplexes, called quatenary complex (" Qplex "), and evaluated DNA delivery efficiency. Qplex is composed with cationic liposome, pDNA, protamine and transferrin. The lipid of liposome M-( α-trimethylammonioacetyl)-didodecyl-D-glutamatechloride (TMAG) / dilauroyl-phospatidylcholine (DLPC) / dioleoylphospatidyl-ethanolamine (DOPE) (1 : 2 : 2). The transfectional efficiency of lac Z gene is increased to 52% with Qplex from 4 % with conventional cationic lipoplexes in Xgal staining. The transfection efficiency is superior in the existence of serum.(5) In order to increase the transfection efficiency in non-viral vector gene therapy, a novel fusogenic peptide, JTS-1, was evaluated in Lipoplex. Luciferase activity in cancer cell lines was greatest in the using JTS-1/LPD complex. The high transfection effeciency was observed by Xgal staining. The mean diameter of LPD is 50 nm and it is so compact compared to lipoplex ( about 600nm ). Fifty percent tumor growth suppression of AsPC-1 cells was shown at the 0.4μg per ml of tob, "a novel tumor suppressor ", DNA plasmid using JTS-1/LPD complex. The suppressive effect was caused to induction of apoptosis. Less

我们进行了肿瘤特异性基因治疗增强BNCT效应的研究。(1)我们制备了聚乙二醇（PEG）结合脂质体包埋的β-<10>B化合物作为给药系统。在注射了μ <10>B-裸脂质体或μ <10>B-PEG-脂质体的小鼠的热中子照射后，相对于对照，AsPC-1肿瘤生长受到抑制。26号结肠肿瘤经转铁蛋白-β<10>-PEG-脂质体BNCT治疗后肿瘤消失。注射转铁蛋白-β<10>-PEG-脂质体和β<10>-B-PEG-脂质体在体内引起热中子照射的最大肿瘤抑制。(2)我们使用CR-39塑料径迹探测器进行中子俘获放射自显影（NCAR）。在立教大学、PSI、约瑟夫·斯特凡研究所或萨克利研究所，将与样品相连的CR-39探测器暴露于热中子或冷中子。PEG<10>-脂质体可以增加肿瘤组织中的B原子的蓄积，从而增加肿瘤组织中的B原子在血液中的滞留， ...更多信息 降低并逃避RES的吞噬作用。(3)我们研制了一种新型的小型脉冲中子发生器。该中子发生器将应用于BNCT。(4)在肿瘤基因治疗中应用非病毒载体时，需要提高转染效率。我们制备了新型的质粒DNA-阳离子脂质复合物，称为四元复合物（“Qplex“），并评估了DNA递送效率。Qplex由阳离子脂质体、pDNA、鱼精蛋白和转铁蛋白组成。脂质体M-（α-三甲基氨乙酰基）-双十二烷基-D-脱乙酰基甜菜碱（TMAG）/二月桂酰磷脂酰胆碱（DLPC）/二油酰磷脂酰乙醇胺（DOPE）（1：2：2）。Xgal染色结果表明，Qplex可使lac Z基因的转染效率从常规阳离子脂质体的4%提高到52%。在血清存在下转染效率上级。(5)为了提高非病毒载体基因治疗的转染效率，在Lipoplex中评价了一种新的融合肽JTS-1。癌细胞系中的荧光素酶活性在使用JTS-1/LPD复合物中最大。Xgal染色显示转染效率高。LPD的平均直径为50 nm，与lipoplex（约600 nm）相比非常紧凑。用JTS-1/LPD复合物，在0.4μg/ml的tob DNA质粒作用下，AsPC-1细胞的肿瘤生长抑制率为50%。其抑制作用是通过诱导细胞凋亡而实现的。少

项目成果

H.Yanagie, K.Ogura, K.Ono, H.Kobayashi et al.: "Application of 1OB entrapped cationic liposome to boron neutron captur therapy for pancreatic cancer model in vivo"Frontiers in Neutron Capture Therapy. 164. 1077-1083 (2001)

H.Yanagie、K.Ogura、K.Ono、H.Kobayashi 等人：“1OB 包埋阳离子脂质体在体内胰腺癌模型硼中子俘获疗法中的应用”中子俘获疗法前沿。

H.Yanagie, K.Ogura, K.Kono, H.Kobayashi et al.: "Accumulation of boron compounds to tumor with polyethylene-glycol binding liposome by using neutron capture autoradiography"J.Appl.Radiation and Isotopes. to be published. (2001)

H.Yanagie、K.Ogura、K.Kono、H.Kobayashi 等人：“通过使用中子捕获放射自显影技术，用聚乙二醇结合脂质体将硼化合物累积到肿瘤”J.Appl.Radiation and Isotopes。

H. Sugiyama, H. Yanagie et al.: "Identification of nuclear export signals in human type C hepatitis virus"FASEB Journal.. 13(7). 1492 (1999)

H. Sugiyama、H. Yanagie 等人：“人丙型肝炎病毒核输出信号的识别”FASEB Journal.. 13(7)。

H. Yanagie, K. Ogura, H. Kobayashi et al.: "Determination of liposomal boron Biodistribution in tumor hearing mice by using neutron capture autoradiography"International Conference nuclear Energy in Central Europe 2001. 616. 2-6 (2001)

H. Yanagie、K. Ogura、H. Kobayashi 等人：“使用中子捕获放射自显影技术测定肿瘤听力小鼠中脂质体硼的生物分布”中欧核能国际会议 2001. 616. 2-6 (2001)

K.Ogura, A.Yamazaki, H.Yanagie et al.: "Neutron capture autoradiography for a study on boron neutron capture therapy"Radiation Measurements. 34. 555-558 (2001)

K.Ogura、A.Yamazaki、H.Yanagie 等人：“中子俘获放射自显影术用于硼中子俘获疗法的研究”辐射测量。