Spectral Imaging of Melanoma Cell Adhesion & Metastasis

黑色素瘤细胞粘附的光谱成像

• 批准号: 6587203
• 负责人: DOROTHEA BECKER
• 金额: $ 33.18万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6587203
• 关键词: antibody; athymic mouse; cadherins; cell adhesion; complementary DNA; connective tissue cells; cyanine; gene expression; gene targeting; human tissue; integrins; melanoma; metastasis; microscopy; monophenol monooxygenase; neoplasm /cancer genetics; neoplasm /cancer invasiveness

DESCRIPTION (provided by applicant): The prognosis for patients with melanoma is directly related to the depth of invasion of the primary lesion at the time of diagnosis. Thus, when diagnosed at an early stage, the patient is often cured by a wide and deep excision of the melanoma. However, once primary melanoma metastasizes to different sites, the prognosis is grave because metastatic melanoma is refractory to conventional therapy. Based upon the finding that compared to melanoma precursor lesions, primary and metastatic human melanomas, and cell lines established from advanced-stage human melanoma specimens, express high levels of the cell adhesion molecules N-cadherin, M-CAM, and beta3 integrin, we postulate that these three genes/proteins, alone or in concert, play important roles in melanoma cell invasion and formation of metastases. To test this hypothesis, we propose to combine antisense gene targeting and optical bioimaging to determine, in vivo, whether these three adhesion molecules govern melanoma cell adhesion, invasion, and metastasis formation. To obtain an answer for this important but unanswered question, primary human melanomas, grown as subcutaneous tumors in nude mice, will be injected, alone and in combination, with vector constructs generating N-cadherin, M-CAM, and/or beta3 integrin antisense transcripts under the control of the human tyrosinase promoter, which is only expressed in human melanoma cells. The tumors will then be injected with different cyanine fluorochrome-conjugated antibodies specific for human N-cadherin, M-CAM, beta3 integrin, and the S100 human melanoma cell surface antigen. Thereupon, non-invasive, macroscopic Spectral Imaging will be performed to document, in vivo, possible loss of cell-cell and cell-stroma interactions in the antisense-targeted melanomas, none of which should be observed in non-targeted control tumors. Following the in vivo imaging analyses of the melanomas, the animals will be sacrificed and tissue sections of their resected tumors and different organs of the nude mice will be subjected to qualitative and quantitative microscopic Spectral Imaging. These ex vivo optical imaging analyses are expected to substantiate the dynamic intratumoral changes captured in vivo, and to provide important information regarding absence versus presence of melanoma metastases in the lungs, abdomen, and brain of mice that carded the antisense-targeted tumors compared to mice that carded the non-targeted tumors.

描述（由申请人提供）：黑色素瘤患者的预后与诊断时原发病灶的浸润深度直接相关。因此，在早期诊断时，患者通常通过广泛而深入的黑色素瘤切除来治愈。然而，一旦原发性黑色素瘤转移到不同的部位，预后是严重的，因为转移性黑色素瘤难以常规治疗。基于我们的发现，与黑色素瘤前体病变、原发性和转移性人类黑色素瘤以及晚期人类黑色素瘤标本建立的细胞系相比，细胞粘附分子N-cadherin、M-CAM和β a3整合素表达水平较高，我们假设这三种基因/蛋白单独或协同在黑色素瘤细胞侵袭和转移形成中发挥重要作用。为了验证这一假设，我们建议将反义基因靶向和光学生物成像结合起来，在体内确定这三种粘附分子是否控制黑色素瘤细胞的粘附、侵袭和转移形成。为了获得这个重要但尚未解决的问题的答案，将在裸鼠皮下生长的原发性人黑色素瘤单独或联合注射，在人类酪氨酸酶启动子的控制下，载体构建产生N-cadherin， M-CAM和/或β a3整合素反义转录物，该启动子仅在人类黑色素瘤细胞中表达。然后将肿瘤注射不同的花青素荧光染料偶联抗体，这些抗体针对人类n -钙粘蛋白、M-CAM、β a3整合素和S100人类黑色素瘤细胞表面抗原。因此，非侵入性的宏观光谱成像将在体内记录反义靶向黑色素瘤中可能失去的细胞-细胞和细胞-基质相互作用，而这些在非靶向对照肿瘤中都不应该观察到。在黑素瘤的体内成像分析之后，将动物处死，对其切除的肿瘤组织切片和裸鼠的不同器官进行定性和定量的显微光谱成像。这些体外光学成像分析有望证实在体内捕获的动态瘤内变化，并提供关于在患有反义靶向肿瘤的小鼠与患有非靶向肿瘤的小鼠的肺、腹部和大脑中黑色素瘤转移的缺失与存在的重要信息。