Control of Eye Melanoma Metastasis

描述（由申请人提供）：该项目的长期目标是明确转移性葡萄膜黑色素瘤到肝脏的病理生物学。这一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，死亡率高达30%。尽管原发肿瘤的治疗有所改善，但死亡率并未下降。葡萄膜黑色素瘤在肝脏形成微转移，肝脏转移性疾病是葡萄膜黑色素瘤患者死亡的主要原因。这些微转移瘤有生长、休眠或渐开线的潜力，这取决于微转移瘤的内在特性和宿主的外在特性。血管抑制素通过调控血管内皮生长因子（VEGF）与色素上皮衍生因子（PEDF）的比值促进黑色素瘤凋亡和维持休眠。干扰素α -2b （IFN）通过促进肝内NK细胞促进微转移性黑色素瘤凋亡。血管抑制素和IFN通过VEGFR1协同作用，VEGFR1在黑色素瘤细胞和NK细胞中表达。第一个目标是控制黑色素瘤细胞系和肝脏微转移环境中PEDF/VEGF的内在比率。这是通过在黑色素瘤细胞系中转染VEGF或PEDF的过表达和siRNA阻断VEGF或PEDF来完成的，用激光捕获显微解剖测定微转移中VEGF和PEDF的mRNA水平，在黑色素瘤/肝细胞共培养中检测VEGF/PEDF，并将肝微转移与肝脏缺氧区联系起来。第二个目的是确定VEGFR1在共培养系统中刺激粘附肝内皮的NK细胞的作用，以及FAS/FAS配体、IFNgamma和/或穿孔素对敲除小鼠NK杀伤的贡献。第三个目标是利用前两个目标中的方法确定血管抑制素/IFN联合控制微转移的机制。

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.