Intra-operative identification of malignant glioma using real-time fluorescence spectroscopic analysis and double staining method

实时荧光光谱分析和双染法术中鉴别恶性胶质瘤

• 批准号: 14571345
• 负责人: KUROIWA Toshihiko
• 金额: $ 1.6万
• 依托单位: Osaka Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571345/
• 关键词: Glioma; Photodynamic diagnosis; Spectroscopic analysis; Photodynamic therapy; Reactive oxygen species; Robotic surgery; 5-ALA; Apoptosis; 腫瘍; 可視化; センサー; 分光解析; 蛍光標識; ロボット

The first purpose of this study is to establish the glioma identification method of PDD using spectroscopic analysis and double staining method. Our second purpose is to develop the next generation PDD agent, which has the excellent tumor specificity. Our third purpose is to achieve the basic studies on the ALA-PDT.To analyze the fluorescence spectrum using fiber-optic spectroscopic tumor sensor, we could identify the tumor specific spectrum. The time resolution of spectroscopic identification is very short, so that this sensing method can be adapted for the sensor type surgical robot system. Next, we developed the new fluorescent agent (FLS-HAS) to conjugate the FLS and human albumin. The FLS-HAS has the excellent tumor specificity compared with 5-ALA. We found that the EPR (electron spin resonance method) could be applicable to the direct detection of ROSs (reactive oxygen species) in the living cell during PDT. The major ROSs of ALA-PDT was singlet oxygen. As for the mechanism of cell death after ALA-PDT, we found that the increases in the activity of both caspase-3 and -9, a decrease in MMP (mitochondrial membrane potential), and a marked increase in cytochrome c in the cytosolic fl-action were found after cells were subjected to PDT. These results indicate that a dysfunction of MMP is followed by mitochondrial cytochrome c release, which triggers apoptosis through a mitochondrial pathway.

本研究的第一个目的是利用光谱分析和双染色法建立PDD的胶质瘤鉴别方法。我们的第二个目的是开发下一代PDD药物，它具有优异的肿瘤特异性。我们的第三个目的是实现ALA-PDT的基础研究。利用光纤光谱肿瘤传感器分析荧光光谱，可以识别肿瘤的特异性光谱。光谱识别的时间分辨率很短，因此该传感方法可以适用于传感器型手术机器人系统。接下来，我们开发了新型荧光剂（FLS-HAS）来将FLS和人白蛋白结合。与5-ALA相比，FLS-HAS具有优异的肿瘤特异性。我们发现EPR（电子自旋共振法）可适用于PDT过程中活细胞中ROS（活性氧）的直接检测。 ALA-PDT的主要ROS是单线态氧。对于ALA-PDT后细胞死亡的机制，我们发现胞浆内caspase-3和-9的活性增加，MMP（线粒体膜电位）降低，细胞色素c显着增加。 -细胞接受PDT后发现作用。这些结果表明，MMP 功能障碍后会导致线粒体细胞色素 c 释放，从而通过线粒体途径触发细胞凋亡。

项目成果

Yoshinaga Kajimoto et al.: "Fiber-optic spectroscopic detection of neoplasm by intraoperative fluorescence labeling"International Congress Series. 1259. 33-38 (2004)

Yoshinaga Kajimoto 等人：“通过术中荧光标记对肿瘤进行光纤光谱检测”国际大会系列。

黒岩敏彦: "蛍光標識手術"Clinical neuroscience. 21. 584-585 (2003)

Toshihiko Kuroiwa：“荧光标记手术”临床神经科学 21. 584-585 (2003)。

Kajimoto Y: "Fiber-optic spectroscopic detection of neoplasm by intraoperative fluorescence labeling"International Congress Series. 1259. 33-38 (2004)

Kajimoto Y：“通过术中荧光标记对肿瘤进行光纤光谱检测”国际大会系列。

Ichioka T: "Enhanced detection of malignant glioma xenograft by fluorescein-human serum albumin conjugate"Journal of Neurooncology. 67. 47-52 (2004)

Ichioka T：“通过荧光素-人血清白蛋白缀合物增强恶性神经胶质瘤异种移植物的检测”神经肿瘤学杂志。

梶本宜永: "光ファイバーを用いた蛍光標識腫瘍センサーの開発.脳腫瘍の研究と治療(田渕和雄編)"九州大学出版会. 561 (2003)

梶本吉永：“利用光纤开发荧光标记肿瘤传感器。脑肿瘤的研究和治疗（田渊和夫编辑）”九州大学出版社561（2003）。