Control of Eye Melanoma Metastsis

描述（由申请人提供）：本项目的长期目标是确定转移性葡萄膜黑色素瘤至肝脏的病理生物学。这些知识将用于控制转移过程，从而降低死亡率。葡萄膜黑色素瘤是最常见的原发性眼内肿瘤，死亡率为30%。尽管原发性肿瘤的治疗有所改善，但死亡率并未降低。葡萄膜黑色素瘤在肝脏中形成微转移，肝脏转移性疾病是葡萄膜黑色素瘤患者死亡的主要原因。这些微转移有可能生长，保持休眠或渐开线，这取决于微转移的内在特性和宿主的外在特性。血管抑素通过调节内源性血管内皮生长因子（VEGF）与色素上皮衍生因子（PEDF）的比例促进黑色素瘤细胞凋亡和维持休眠。干扰素α-2b（IFN）通过增强内在肝NK细胞促进微转移性黑色素瘤细胞凋亡。血管抑素和IFN通过黑素瘤细胞和NK细胞表达的VEGFR 1协同作用。第一个目的是操纵黑色素瘤细胞系和肝脏微转移环境中的固有PEDF/VEGF比率。这是通过转染VEGF或PEDF的过表达和siRNA阻断黑素瘤细胞系中VEGF或PEDF，用激光捕获显微切割测定微转移中的VEGF和PEDF mRNA水平，测试黑素瘤/肝细胞共培养物中的VEGF/PEDF，并将肝微转移与肝脏中的缺氧区相关联来完成的。第二个目的是确定VEGFR 1刺激NK细胞在共培养系统中粘附于肝内皮的作用，以及FAS/FAS配体、IFN γ和/或穿孔素对敲除小鼠NK杀伤的贡献。第三个目标确定了使用前两个目标中的方法的组合血管抑素/IFN控制微转移的机制。

DESCRIPTION (provided by applicant): The long-term goal of this project is to define the pathobiology of metastatic uveal melanoma to the liver. This knowledge will be used to control the metastatic process thus decreasing mortality. Uveal melanoma is the most common primary intraocular tumor and has an associated 30% mortality. This mortality rate has not decreased, despite improvements in treating the primary tumor. Uveal melanoma forms micrometastases in the liver and metastatic disease of the liver is the leading cause of death in patients with uveal melanoma. These micrometastases have the potential to grow, remain dormant, or involute, depending on intrinsic properties of the micrometastases and extrinsic properties of the host. Angiostatin promotes melanoma apoptosis and maintenance of dormancy by manipulating the intrinsic vascular endothelial growth factor (VEGF) to pigment epithelium derived factor (PEDF) ratio. Interferon alpha-2b (IFN) promotes micrometastatic melanoma apoptosis by boosting intrinsic hepatic NK cells. Angiostatin and IFN act synergistically via VEGFR1 which is expressed by melanoma cells and NK cells. The first aim is to manipulate the intrinsic PEDF/VEGF ratio in melanoma cell lines and within the micrometastatic environment in the liver. This is done by transfection for overexpression of VEGF or PEDF and siRNA blocking of VEGF or PEDF in melanoma cell lines, determining VEGF and PEDF mRNA levels in the micrometastases with laser capture microdissection, testing for VEGF/PEDF in co-cultures of melanoma/hepatocytes, and correlating hepatic micrometastases with hypoxic zones in the liver. The second aim determines the role of VEGFR1 stimulation of NK cells with adherence to hepatic endothelium in a coculture system and the contribution of FAS/FAS ligand, IFNgamma, and/or perforin to NK killing in knockout mice. The third aim determines the mechanism of combined angiostatin/IFN control of the micrometastases using methods in the first two aims.