Experimental Uveal Melanoma and Ocular Immune Privilege

实验性葡萄膜黑色素瘤和眼部免疫特权

• 批准号: 7030002
• 负责人: KYLE C MCKENNA
• 金额: $ 7.65万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-01 至 2007-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7030002
• 关键词: disease /disorder model; eye neoplasms; genetically modified animals; green fluorescent proteins; immune response; laboratory mouse; liver neoplasms; melanoma; metastasis; model design /development; neoplasm /cancer immunology; neoplastic growth; neoplastic transformation; recombinase; simian virus 40; tamoxifen; tissue /cell culture; transfection /expression vector; tumor antigens; uvea

DESCRIPTION: Uveal melanoma is the most common primary malignant intraocular tumor in adults. Metastatic disease of the liver is the leading cause of death in uveal melanoma patients and there is currently no effective treatment. Developing and testing therapies for uveal melanoma has been hampered by the absence of a satisfactory animal model in which uveal melanocytes are transformed in situ and metastasize to the liver. In addition, few murine melanoma cell lines of ocular origin are available for studies aimed at understanding the mechanisms of immune evasion and liver metastasis by uveal melanoma. The objective of this proposal is to develop an SV40 T-antigen (Tag) transgene with elements that allow inducible expression of SV40 Tag by the tyrosinase promoter in the presence of Cre-recombinase. This transgene will permit inducible in situ transformation of pigmented tissues, including melanocytes, in C57BI/6 mice, by targeted drug delivery into the anterior or posterior segments of the eye to induce Cre-recombinase activity. These mice will be used to characterize the immune response to primary and metastatic uveal melanoma by monitoring SV40 Tag-specific immune responses. The immune response to SV40 Tag is well characterized in C57BI/6 mice and tetramers composed of soluble MHC Class I : SV40 Tag peptides are available along with an SV40 Tag specific TCR transgenic strain for studies aimed at determining the fate of CD8+ tumor-specific T cells in primary and metastatic uveal melanoma. In addition, these SV40 Tag transgenic mice will be a source for generating uveal melanoma cell lines. As the tyrosinase promoter is also active in cutaneous melanocytes, this transgenic mouse strain may also serve as a model of cutaneous melanoma which is inducible by drug delivery to the skin. Comparison of uveal and cutaneous melanoma cell lines derived from SV40 Tag transgenic mice may reveal novel molecules that control immune evasion and liver metastasis associated with uveal but not cutaneous melanoma. Understanding the mechanisms of immune evasion by uveal melanomas may lead to therapeutic approaches to promote tumor elimination. Furthermore, manipulation of these immune-suppressive mechanisms may be employed to inhibit T cell responses to prolong graft transplantation in the eye, for example, RPE transplants, or mitigate T cell mediated autoimmune uveitis.

描述：卵子黑色素瘤是成年人最常见的原发性恶性肿瘤内肿瘤。肝脏的转移性疾病是紫veal黑色素瘤患者死亡的主要原因，目前尚无有效的治疗。缺乏令人满意的动物模型，在原位转化了紫veal虫黑色素细胞并向肝脏转移，因此无法开发和测试紫美氏瘤的疗法。此外，几乎很少有眼部的鼠类黑色素瘤细胞系可用于研究旨在理解紫葡萄酸黑色素瘤免疫逃避和肝转移的机制的研究。该提案的目的是开发SV40 T抗原（TAG）转基因，其元素允许在存在CRE - 成分酶的情况下通过酪氨酸酶启动子诱导的SV40 TAG表达。该转基因将允许在C57BI/6小鼠中通过将药物递送到眼睛的前部或后段来诱导Cre-cobinase酶活性，从而允许在C57BI/6小鼠中进行色素组织的原位转化。这些小鼠将通过监测特定于SV40 TAG特异性来表征对原发性和转移性卵子黑色素瘤的免疫反应 免疫反应。 C57BI/6小鼠的免疫反应对SV40标签的特征很好，并且 由可溶性MHC I类组成的四聚体：SV40标签肽以及SV40标签 针对旨在确定原发性和转移性卵子黑色素瘤中CD8+肿瘤特异性T细胞的命运的研究的特异性TCR转基因菌株。另外，这些SV40 TAG转基因小鼠将是产生紫veal黑色素瘤细胞系的来源。由于酪氨酸酶启动子也在皮肤黑素细胞中活跃，因此这种转基因小鼠菌株也可以作为皮肤黑色素瘤的模型，可通过药物递送到皮肤中诱导。从SV40 TAG转基因小鼠衍生的紫菜和皮肤黑色素瘤细胞系的比较可能揭示出可控制免疫逃避和肝转移的新型分子，但与紫veal虫相关，但不是皮肤黑色素瘤。了解卵子黑色素瘤免疫逃避的机制可能导致促进肿瘤消除的治疗方法。此外，可以采用对这些免疫抑制机制的操纵来抑制T细胞对眼睛长期移植移植的反应，例如，RPE移植或减轻T细胞介导的自身免疫性葡萄膜炎。