Development of comprehensive methods for diagnosis/therapy by optic bio-imaging

在本研究中，我们进行了两个主要实验。主题A)用于肿瘤和药物敏感性定性/定量评价的生物成像方法的发展：基于荧光素酶的新型光学证明在体内分子功能。我们开发了检测caspase-3活性的光学探针，并对其进行腺病毒载体编码。利用这种方法，我们进行了细胞和小鼠实验，成功地显示了组织中的凋亡。特别是在缺氧/再氧合（缺血/再灌注）和肝切除术后肝再生中，我们成功地成像了氧化应激诱导的肝细胞凋亡损伤的动态。主题B)利用新开发的光学探针（体内/体外成像）检测和表征肿瘤的新诊断/治疗方法的发展。我们将光学探针安装到各种抗体上，包括Dlk-1/CEA/EMA/Ep-CAM。通过FACS/Western Blot检测不同细胞系中Dlk-1/CEA/EMA/Ep-CAM的表达，并对nu/nu小鼠进行皮下肿瘤细胞植入。在表达Dlk-1的肝癌细胞系Huh-7的实验中，我们证明了这种新型光学探针在体内的有效性。我们在没有紫外线照射的情况下，测量了小鼠植入式肝肿瘤和皮下肿瘤的足够信号。我们现在正在挑战其他抗体以确保临床相关性。

In the present study, we performed two major experiments.Theme A) Development of bio-imaging method for qualitative/quantitative evaluation of tumors and drug sensitivity : luciferase-based new optic prove for in vivo molecular function. We developed optic probe for caspase-3 activity and adenovirus vector encoding for the probe. By using this, we performed cellular and mice experiments, successfully showing apoptosis in the tissue. Especially, in hypoxia/reoxygenation (ischemia/reperfusion) and post-hepatectomy liver regeneration, we succeeded in imaging the dynamism of oxidative stress-induced apoptotic liver injury.Theme B) Development of a new diagnostic/therapeutic method for tumor detection and characterization by the newly developed optic probe (in vivo/ex vivo imaging). We mounted the optic probe to various antibodies including Dlk-1/CEA/EMA/Ep-CAM. Also, we checked expressions of Dlk-1/CEA/EMA/Ep-CAM in various cell lines by FACS/Western Blot, and performed subcutaneous tumor cell implantation in nu/nu mice. In the experiments using hepatoma cell line (Huh-7) expressing Dlk-1, we proved the efficacy of our new optic probe in vivo. We measured enough signals from implanted liver tumor as well as subcutaneous tumor in mice without UV exposure. We are now challenging other antibodies to ensure clinical relevance.