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

基本信息

批准号：
9822110
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
项目类别：
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/9822110
关键词：
Affinity Chromatography Affinity Chromatography Area Area Asparagine Asparagine Bar Codes Biological Biological Biological Assay Biological Assay Biological Models Biological Models Biology Biology Brain Brain Cancer Biology Cancer Biology Cancer Patient Cancer Patient Carbohydrates Carbohydrates Cardiovascular system Cardiovascular system Cell Adhesion Cell Adhesion Cell Adhesion Molecules Cell Adhesion Molecules Cell surface Cell surface Cells Cells Cessation of life Cessation of life Clinical Clinical Complex Complex Coupling Coupling Disseminated Malignant Neoplasm Disseminated Malignant Neoplasm Distal Distal Distant Distant Enzymes Enzymes Epitopes Epitopes Fluorescence Fluorescence Fucosyltransferase Fucosyltransferase Genetic Transcription Genetic Transcription Glycoproteins Glycoproteins Glycoside Hydrolases Glycoside Hydrolases Homing Homing Immune Immune Immune Evasion Immune Evasion In Vitro In Vitro Invaded Invaded Investigation Investigation Knowledge Knowledge Laboratories Laboratories Lectin Lectin Libraries Libraries Link Link Liver Liver Lung Lung Lymphatic System Lymphatic System Malignant - descriptor Malignant - descriptor Malignant Neoplasms Malignant Neoplasms Malignant neoplasm of prostate Malignant neoplasm of prostate Mediating Mediating Melanoma Cell Melanoma Cell Metastatic Melanoma Metastatic Melanoma Metastatic malignant neoplasm to brain Metastatic malignant neoplasm to brain MicroRNAs MicroRNAs Movement Movement Mutate Mutate Neoplasm Metastasis Neoplasm Metastasis Neural Cell Adhesion Molecule L1 Neural Cell Adhesion Molecule L1 Non-Small-Cell Lung Carcinoma Non-Small-Cell Lung Carcinoma Organ Organ Pathway interactions Pathway interactions Patients Patients Pattern Pattern Pigments Pigments Polysaccharides Polysaccharides Primary Neoplasm Primary Neoplasm Process Process Prognosis Proliferating Proliferating Property Property Protein Glycosylation Protein Glycosylation Proteins Proteins Regulation Regulation Research Research Role Role Sampling Sampling Secondary to Secondary to Series Series Serine Serine Shotguns Shotguns Signal Pathway Signal Pathway Site Site Skin Skin Specificity Specificity Structure Structure Systems Biology Systems Biology Therapeutic Therapeutic Threonine Threonine Tissue Sample Tissue Sample Tissue Survival Tissue Survival Tropism Tropism Validation Validation Work Work Xenograft Model Xenograft Model angiogenesis angiogenesis bone bone cancer cell cancer cell cancer type cancer type candidate validation candidate validation capillary bed capillary bed cell motility cell motility clinically relevant clinically relevant data mining data mining differential expression differential expression glycoproteomics glycoproteomics glycosylation glycosylation glycosyltransferase glycosyltransferase high throughput screening high throughput screening in vitro Assay in vitro Assay in vitro Model in vitro Model in vivo in vivo in vivo Model in vivo Model innovation innovation insight insight liquid chromatography mass spectrometry liquid chromatography mass spectrometry lymph nodes lymph nodes malignant phenotype malignant phenotype melanocyte melanocyte melanoma melanoma metastatic process metastatic process mortality mortality neoplastic cell neoplastic cell novel therapeutics novel therapeutics outcome forecast programs programs screening screening small hairpin RNA small hairpin RNA treatment site treatment site tumor tumor tumor progression tumor progression

项目摘要

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.

糖基化作为黑色素瘤转移向性的调节剂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。肿瘤转移至继发部位是 90% 癌症死亡的原因。一个显着特点是转移是原发肿瘤定植次要器官的能力。这促使人们寻求确定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（例如，脑、肺、肝）。黑色素瘤最具破坏性的并发症之一是大约 50% 的患者患有转移性黑色素瘤会发生脑转移，大多数患者存活时间不到 6 个月。患者患有脑转移的患者无法从新疗法中受益，并且预后极差。了解大脑和其他器官适应的基础可能会揭示新的疗法。碳水化合物，其变化肿瘤参与免疫逃避、细胞归巢至组织、存活和锚定。我们最近的临床相关研究确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次级位点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方法来确定治疗相关性聚糖结构和糖基化酶作为抗转移治疗的靶标。我将确定候选人通过相关体内模型的聚糖分析来调节聚糖和糖原的位点特异性转移以及带有条形码的糖原混合shRNA文库的高通量筛选。候选聚糖和糖原将通过黑色素瘤患者 FFPE 样本的凝集素荧光和 IHC 分析进行验证。我们将使用体内异种移植模型进一步验证候选糖原并剖析其作用机制通过各种体外测定。此外，我们将使用糖蛋白组策略，耦合凝集素亲和力使用 LC/MS 鸟枪法蛋白质鉴定进行纯化，以鉴定具有特定聚糖的糖基化蛋白质主题。最后，将研究与位点特异性转移相关的糖蛋白的机制行动。积极参与黑色素瘤趋向性的聚糖和糖基化酶的鉴定以及对其作用机制的精确理解有可能提供大量聚糖表位和酶尚未被探索作为治疗位点特异性转移的抗肿瘤靶标。