Analysis of Glycomic Regulators in Melanoma Progression

项目摘要 癌症的一个显著特征是细胞糖基化的改变。本地化的转变 癌症到转移性疾病，这通常会导致不良的预后，关键的特点是明显的变化 糖基化。转移性癌细胞合成糖链异常水平和结构变化的比较 与他们的非转移性对应物。转移相关的糖链可以改变细胞的活动，例如 黏附、侵袭和增殖，以及促进转移前凝集素的结合。因此， 转移相关的多聚糖及其相关的多聚糖合成酶为肿瘤提供了有吸引力的靶点 癌症进展的治疗干预。根据这种协同UO1肿瘤糖组分 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 表面多聚糖和肿瘤糖生物学研究糖链结构的修饰如何影响 恶性进展。我们的指导性假设是获得不同的转移相关糖链 影响恶性行为和转移潜能。在初步研究中，基因集浓缩分析 对常见人类癌症类型进行的研究表明，转移性黑色素瘤最重要的是 富含‘Glycome’基因签名。随后的血糖分析表明转移性黑色素瘤 细胞独一无二地显示出高水平的I-线性聚N-乙酰乳糖胺(Polylacs)，而正常人 黑素细胞几乎只表达I-分支聚乳酸。进一步比较糖类基因图谱 在正常黑素细胞和转移性黑色素瘤细胞之间发现i-分支1，6的抑制水平 转移性黑色素瘤细胞中N-乙酰氨基葡萄糖基转移酶(GCNT2)的表达 黑色素瘤进展。GCNT2在人黑色素瘤模型中的强制表达强烈表明，I- BRANCH/GCNT2可显著抑制体内黑色素瘤生长和细胞信号转导 细胞增殖与胰岛素样生长因子-1受体和β1整合素相关。I-分支GCNT2活性 还抑制了促黑色素瘤转移凝集素、Galectin(Gal)-3的结合。在这笔赠款中，我们将利用这些 激动人心的黑色素瘤血糖数据研究I-线性/I-支链聚乳酸和GCNT2作为调节因子的作用 转移性黑色素瘤的行为。具体目的是：(1)分析I-支链多糖的功能 和GCNT2对黑色素瘤细胞活性的影响(2)确定I-BRANCH/GCNT2是否表达 预测黑色素瘤的发展。这些目标将由一个由肿瘤专家组成的合作团队来探索 糖生物学、糖分析和黑色素瘤病理学，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。 我们深远的目标是了解黑色素瘤是如何转移的--黑色素瘤致死的原因 病人。重要的是，我们的发现将为如何针对血糖调节剂提供新的见解 治疗开发或用作预测患者黑色素瘤转移和临床结果的生物标志物。

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.