Glycosylation as a Regulator of Tropism of Melanoma Metastasis - Resubmission - 1

糖基化作为黑色素瘤转移的趋向性调节因子 恶性黑色素瘤是一种起源于黑素细胞的癌症，黑素细胞是皮肤的色素细胞。 肿瘤转移到继发部位是导致90%癌症死亡的原因。的一个显著特点 转移是指原发肿瘤定植于次级器官的能力。这促使了一种寻找身份的探索 支持黑色素瘤转移到特定器官作为继发部位的因素和机制(如： 脑、肺、肝)。黑色素瘤最具破坏性的并发症之一是大约50%的患者 转移性黑色素瘤发生脑转移，大多数患者存活不到6个月。病人 有脑转移的患者不能从新的治疗中受益，预后极差。了解 根据大脑和其他器官的适应可能会揭示新的治疗方法。碳水化合物，它在 肿瘤，参与免疫逃逸、细胞归巢到组织、生存和锚定。我们最近 临床相关研究确定了原发和转移性黑色素瘤不同的糖基化模式。 此外，我们对患者样本的初步研究表明，特定的糖基化模式是一个位点- 转移的特异性。 我假设，肿瘤细胞对不同次级部位的适应需要细胞的特定变化 表面糖基化。我提议的工作将使用创新的方法来确定与治疗相关的 糖链结构和糖基化酶作为抗转移治疗的靶点。我会找出候选人 糖链和糖基因通过相关体内模型的糖链图谱调节部位特异性转移 以及高通量筛选，带有条形码池的糖基因shRNA文库。候选多糖和 糖基因将通过凝集素荧光和黑色素瘤患者FFPE样本的IHC分析来验证。我们 将使用体内异种移植模型进一步验证候选糖基因并剖析其作用机制 通过各种体外试验。此外，我们将使用糖蛋白组学策略，偶联凝集素亲和力 LC/MS鉴定蛋白质的纯化，以鉴定具有特定糖基化的蛋白质 Motif。最后，我们将研究特定部位转移相关糖蛋白的作用机制。 行动。积极参与黑色素瘤趋向性的糖链和糖基化酶的鉴定 此外，对其作用机制的精确了解有可能提供大量的葡聚糖 表位和酶，这是尚未被发现的抗肿瘤靶点，用于治疗特定部位转移。

Glycosylation as a regulator of tropism of melanoma metastasis Malignant melanoma is a type of cancer arising from melanocytes, the pigmented cells of the skin. Metastases of tumors to secondary sites are the cause of 90% of cancer mortality. A salient feature of metastasis is the ability of a primary tumor to colonize secondary organs. This has prompted a quest to identify the factors and mechanisms that support melanoma metastasis to specific organs as secondary sites (e.g. brain, lung, liver). One of the most devastating complications of melanoma is that around 50% of patients with metastatic melanoma develop brain metastasis, after which most patients survive less than 6 months. Patients with brain metastasis don't benefit from new therapies and have extremely poor prognosis. Understanding the basis to brain and other organs adaptation may reveal new therapies. Carbohydrates, which are altered in tumors, are involved in immune evasion, homing of cells to tissues, survival, and anchorage. Our recent clinically relevant study identified distinct glycosylation patterns of primary and metastatic melanoma. Moreover, our preliminary studies of patient samples suggest that specific glycosylation patterns are a site- specific feature of metastasis. I hypothesize that adaptation of tumor cells to different secondary sites requires specific changes in cell surface glycosylation. My proposed work will use innovative approaches to identify therapeutically relevant glycan structures and glycosylation enzymes as targets for anti-metastatic therapies. I will identify candidate glycans and glycogenes to regulate site-specific metastasis through glycan profiling of relevant in vivo models and high-throughput screens with a barcoded pooled shRNA library of glycogenes. Candidate glycans and glycogenes will be validated by lectin fluorescence and IHC analysis of melanoma patient FFPE samples. We will further validate candidate glycogenes using in vivo xenograft models and dissect their mechanism of action through various in vitro assays. Further, we will use a glycoproteomic strategy, coupling lectin-affinity purification with LC/MS shotgun protein identification, to identify glycosylated proteins with a specific glycan motif. Finally, site-specific metastasis related glycoproteins would be investigated for their mechanism of action. The identification of glycans and glycosylation enzymes actively participating in melanoma tropism as well as a precise understanding of their mechanism of action has the potential to provide a trove of glycan epitopes and enzymes, that are unexplored as anti-tumor targets for the treatment of site-specific metastases.