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

糖基化作为黑色素瘤转移倾向的调节剂 - 重新提交 - 1

• 批准号: 10474953
• 负责人: Praveen Agrawal
• 金额: $ 19.44万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10474953
• 关键词: Affinity Chromatography; Area; Asparagine; Bar Codes; Biological; Biological Assay; Biological Models; Biology; Brain; Cancer Biology; Cancer Patient; Carbohydrates; Cardiovascular system; Cell Adhesion; Cell Adhesion Molecules; Cell surface; Cells; Cessation of life; Clinical; Complex; Coupling; Disseminated Malignant Neoplasm; Distal; Distant; Enzymes; Epitopes; Fluorescence; Fucosyltransferase; Genetic Transcription; Glycoproteins; Glycoside Hydrolases; Homing; Immune; Immune Evasion; In Vitro; Invaded; Investigation; Knowledge; Laboratories; Lectin; Libraries; Link; Liver; Lung; Lymphatic System; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of prostate; Mediating; Melanoma Cell; Metastatic Melanoma; Metastatic malignant neoplasm to brain; MicroRNAs; Movement; Mutate; Neoplasm Metastasis; Neural Cell Adhesion Molecule L1; Non-Small-Cell Lung Carcinoma; Organ; Pathway interactions; Patients; Pattern; Pigments; Polysaccharides; Primary Neoplasm; Process; Prognosis; Proliferating; Property; Protein Glycosylation; Proteins; Regulation; Research; Role; Sampling; Secondary to; Series; Serine; Shotguns; Signal Pathway; Site; Skin; Specificity; Structure; Systems Biology; Therapeutic; Threonine; Tissue Sample; Tissue Survival; Tropism; Validation; Work; Xenograft Model; angiogenesis; bone; cancer cell; cancer type; candidate validation; capillary bed; cell motility; clinically relevant; data mining; differential expression; glycoproteomics; glycosylation; glycosyltransferase; high throughput screening; in vitro Assay; in vitro Model; in vivo; in vivo Model; innovation; insight; liquid chromatography mass spectrometry; lymph nodes; malignant phenotype; melanocyte; melanoma; metastatic process; mortality; neoplastic cell; novel therapeutics; programs; screening; small hairpin RNA; treatment site; tumor; tumor progression

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.

糖基化对黑色素瘤转移趋向性的调节作用 恶性黑色素瘤是一种由黑色素细胞（皮肤的色素细胞）引起的癌症。 肿瘤转移到继发部位是90%癌症死亡的原因。的一个显著特点 转移是原发性肿瘤定植于继发性器官的能力。这促使人们寻求确定 支持黑色素瘤转移到作为次要部位的特定器官的因素和机制（例如， 脑、肺、肝）。黑色素瘤最具破坏性的并发症之一是，大约50%的黑色素瘤患者， 转移性黑色素瘤发展为脑转移，之后大多数患者存活不到6个月。患者 患有脑转移的患者不能从新的治疗中获益，并且预后极差。了解 根据大脑和其他器官的适应可能会发现新的治疗方法。碳水化合物，它们在 肿瘤参与免疫逃避、细胞归巢至组织、存活和锚定。我们最近 临床相关研究鉴定了原发性和转移性黑色素瘤的不同糖基化模式。 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- 转移的具体特征。 我假设肿瘤细胞对不同继发部位的适应需要细胞内特定的变化， 表面糖化我建议的工作将使用创新的方法来确定治疗相关的 聚糖结构和糖基化酶作为抗转移治疗的靶点。我将确定候选人 通过相关体内模型的聚糖谱分析来调节位点特异性转移的聚糖和糖基因 以及用糖基因的条形码化的汇集的shRNA文库进行高通量筛选。候选聚糖和 将通过黑色素瘤患者FFPE样品的凝集素荧光和IHC分析来验证糖基因。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，我们将使用糖蛋白质组学策略， 用LC/MS鸟枪蛋白鉴定纯化，以鉴定具有特定聚糖的糖基化蛋白 母题最后，将研究位点特异性转移相关糖蛋白的作用机制。 行动上鉴定积极参与黑色素瘤嗜性的聚糖和糖基化酶， 以及对其作用机制的精确理解有可能提供大量的聚糖 表位和酶，其未被探索为用于治疗位点特异性转移的抗肿瘤靶标。