Analysis of Glycomic Regulators in Melanoma Progression

项目摘要 癌症的标志性特征之一是细胞糖基化的改变。本地化的过渡 癌症转移性疾病，这通常导致预后不良，关键的特点是专利的变化， 糖基化转移性癌细胞合成异常水平和可变结构的聚糖， 与非转移性的对应物。转移相关聚糖可以改变细胞活性，例如 粘附、侵袭和增殖，以及促进促转移凝集素的结合。因此，我们认为， 转移相关的聚糖及其相关的聚糖合成酶提供了有吸引力的靶点， 癌症进展的治疗干预。根据这一合作UO 1肿瘤糖组学 指令，我们寻求利用我们在细胞分析方面的互补专业知识和实验室进展， 表面聚糖和肿瘤糖生物学，以检查聚糖结构的修饰如何影响 恶性进展我们的指导假设是，获得不同的转移相关聚糖 影响恶性行为和转移潜力。在初步研究中，基因集富集分析 在9种常见的人类癌症类型中进行的研究表明，转移性黑色素瘤，首先， 富含“糖组”基因标记。随后的糖组学分析表明转移性黑色素瘤 细胞独特地显示高水平的i-线性聚-N-乙酰基乳糖胺（polylacs），而正常人 黑素细胞几乎只表达I-分支多聚乳酸。进一步比较糖组基因谱 在正常黑素细胞和转移性黑色素瘤细胞之间鉴定出I分支β 1，6 转移性黑色素瘤细胞中的N-乙酰葡糖胺转移酶（GCNT 2），与 黑素瘤进展。在人黑色素瘤模型中增强的GCNT 2表达强烈表明I- 分支/GCNT 2导致体内黑色素瘤生长和细胞信号传导显着减弱， 与胰岛素样生长因子-1受体和β 1整联蛋白相关的增殖。I-分支GCNT 2活性 还抑制促黑素瘤转移凝集素半乳糖凝集素（Gal）-3的结合。在这次资助中，我们将利用这些 令人兴奋的黑色素瘤糖组学数据，以研究i-线性/I-分支聚乳酸和GCNT 2作为调节剂的作用 转移性黑色素瘤的行为具体目的是：（1）分析I-支链聚糖的功能 和GCNT 2对黑色素瘤细胞活性的影响以及（2）为了确定I-分支/GCNT 2表达是否 预测黑色素瘤的进展这些目标将由一个肿瘤专家合作小组进行探索 糖生物学、糖分析和黑色素瘤病理学，并且包括使用临床相关的人类和 小鼠黑色素瘤模型和创新的组织病理学，糖分析和基因调控系统。 我们的长远目标是了解黑色素瘤如何转移-黑色素瘤致死的因果步骤， 患者重要的是，我们的研究结果将为糖组学调节因子如何被靶向提供新的见解。 治疗开发或用作生物标志物以预测患者中的黑素瘤转移和临床结果。

PROJECT ABSTRACT One of the hallmark features of cancer is the alteration of cellular glycosylation. The transition of localized cancer to metastatic disease, which commonly leads to poor prognosis, critically features patent changes in glycosylation. Metastatic cancer cells synthesize aberrant levels and variable structures of glycans compared with their non-metastatic counterparts. Metastasis-associated glycans can modify cellular activities, such as adhesion, invasion and proliferation, as well as encourage binding of pro-metastatic lectins. Hence, metastasis-associated glycans and their related glycan-synthesizing enzymes offer attractive targets for therapeutic interference of cancer progression. In accordance with this collaborative UO1 tumor glycomics directive, we seek to leverage our complementary expertise and laboratory advances on analytics of cell surface glycans and on tumor glycobiology to examine how modifications in glycan structure influence malignant progression. Our guiding hypothesis is that acquisition of distinct metastasis-associated glycans impacts malignant behavior and metastatic potential. In preliminary studies, gene set enrichment analysis performed across (9) common human cancer types indicated that metastatic melanomas, above all, were enriched for a `glycome' gene signature. Subsequent glycomic analysis indicated that metastatic melanoma cells uniquely displayed a high level of i-linear poly-N-acetyllactosamines (polylacs), whereas normal human melanocytes almost exclusively expressed I-branched polylacs. Further comparative glycome gene profiling between normal melanocytes and metastatic melanoma cells identified suppressed levels of I-branching ß1,6 N-acetylglucosaminyltransferase (GCNT2) in metastatic melanoma cells that was significantly correlated with melanoma progression. Enforced GCNT2 expression in human melanoma models strongly indicated that I- branch/GCNT2 caused significant attenuation of melanoma growth in vivo and of cell signaling and proliferation associated with insulin-like growth factor-1 receptor and ß1 integrins. I-branching GCNT2 activity also inhibited binding of pro-melanoma metastasis lectin, galectin (Gal)-3. In this grant, we will leverage these exciting melanoma glycomic data to investigate the role of i-linear/I-branch polylacs and GCNT2 as regulators of metastatic melanoma behavior. The Specific Aims are: (1) To analyze function of I-branched glycans and GCNT2 on melanoma cell activity and (2) To determine whether I-branch/GCNT2 expression predicts melanoma progression. These Aims will be explored by a collaborative team of experts in tumor glycobiology, glyco-analytics and melanoma pathology and include the use of clinically-relevant human and mouse melanoma models and innovative histo-pathological, glyco-analytical and gene regulation systems. Our far-reaching goal is to understand how melanomas metastasize - the causal step of melanoma lethality in patients. Importantly, our findings will offer new insights into how glycomic regulators can be targeted for therapeutic exploitation or used as biomarkers to predict melanoma metastasis and clinical outcome in patients.