Targeted Isolation and Identification of Sialylated Glycoproteins in Cancer Tissues, Cells and Biofluids

癌症组织、细胞和生物液中唾液酸化糖蛋白的靶向分离和鉴定

基本信息

批准号：
10358217
负责人：
RICHARD R. DRAKE
金额：
$ 33.7万
依托单位：
MEDICAL UNIVERSITY OF SOUTH CAROLINA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-03-15 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10358217
关键词：
Affinity Alkynes Antibodies Antigens Area Azides Biological Markers CA-19-9 Antigen Cancer Diagnostics Cancer Patient Cancer cell line Carbohydrates Cell Communication Cell Line Cell surface Cell-Cell Adhesion Cells Chemistry Clinical Complex Detection Development Drug or chemical Tissue Distribution Environment Evaluation Extracellular Matrix FDA approved Formalin Fucose Glycocalyx Glycopeptides Glycoprotein Degradation Pathway Glycoproteins Goals Histopathology Human Human Resources Image Immune Individual Isomerism Knowledge Laboratories Ligand Binding Link Malignant Neoplasms Malignant neoplasm of pancreas Malignant neoplasm of prostate Maps Mediating Methodology Microscopy Neoplasm Metastasis Patients Peptide Hydrolases Plasma Polysaccharides Post-Translational Protein Processing Process Prostatic Proteins Proteoglycan Proteomics Reagent Receptor Signaling Reproducibility Research Resolution Sampling Serum Sialic Acids Site Solid Neoplasm Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Surface Therapeutic Tissue imaging Tissues Tumor Antigens Tumor Biology Tumor Markers Tumor Tissue Tumor-Derived Urine amidation biobank cancer biomarkers cancer prevention carcinogenesis cell stroma chemical reaction clinically relevant extracellular glycosylation human tissue imaging approach improved interest mass spectrometric imaging neoplastic cell novel novel diagnostics pancreatic cancer cells portability prognosis biomarker sialyl Lewis a sialyl Lewis x tool tumor tumor microenvironment

项目摘要

Glycoproteins account for the majority of the proteins located at the cell surface and in the extracellular environment. During carcinogenesis the normal function of these glycoproteins as binding ligands for cell-cell adhesion, extracellular matrix molecules, signaling receptors, and immune cells becomes altered. Many of the steps in metastatic progression involve re-organization and degradation of the glycoproteins in the extracellular matrix. Hence, glycan and glycoprotein antigens still comprise the majority of FDA approved diagnostic cancer biomarkers. Our group has recently developed multiple N-linked glycan imaging mass spectrometry workflows to characterize the glycomes in clinically relevant tumor tissues, biofluids and cells, as well as individually targeted glycoproteins by immunocapture. In this proposal, application of a new click chemistry reagent specific for targeting of 2,3 sialic acid antigens, like sialyl Lewis A/CA19-9, sialyl Lewis X and sTRA, will be used to improve isolation and sequencing of tumor-associated glycopeptides from FFPE tissues, biofluids and cells. Standard proteases and new ECM-targeted proteases will be used in the analyses. To accomplish this, three Specific Aims are proposed: 1. Isolate, identify and co-localize sialic acid glycoproteins from clinical FFPE cancer tissues; 2. Isolate and identify sialic acid glycoproteins in plasma cancer samples; 3. Isolate and identify sialic acid glycoproteins in cancer cell lines with high levels of 2,3 sialic antigens. The carbohydrate-targeted click capture reagents developed can be applied directly to any FFPE tissue, cell or biofluid of interest using simple chemical reactions. The combination of tools developed herein will facilitate the identification of specific glycopeptide and glycoprotein species in relevant cancer samples which can be applied across the spectrum of cancer prevention, progression, prognosis, and biomarker applications.

糖蛋白解释了位于细胞表面和中的大多数蛋白质细胞外环境。在癌变期间，这些糖蛋白的正常功能作为结合配体，用于细胞细胞粘附，细胞外基质分子，信号受体和免疫细胞变化。转移进展的许多步骤涉及重组和细胞外基质中糖蛋白的降解。因此，聚糖和糖蛋白抗原仍然包括大多数FDA批准的诊断癌生物标志物。我们的小组最近已经开发了多个N-连接的聚糖成像质谱工作流程表征临床相关的肿瘤组织，生物流体和细胞中的聚糖以及通过免疫接收将单独靶向糖蛋白。在此提案中，应用新点击化学试剂针对靶向2,3唾液酸抗原的特异 Siallyl Lewis X和Stra将用于改善与肿瘤相关的分离和测序来自FFPE组织，生物流体和细胞的糖磷酸酯。标准蛋白酶和新的ECM靶向蛋白酶将用于分析。为此，提出了三个具体目标：1。分离，鉴定和鉴定并共定位于临床FFPE癌组织中的唾液酸糖蛋白； 2。分离并鉴定血浆癌样品中的唾液酸糖蛋白； 3。孤立并识别高水平2,3唾液抗原的癌细胞系中的唾液酸糖蛋白。这开发的碳含碳水化合物的点击捕获试剂可以直接应用于任何FFPE组织，使用简单的化学反应的细胞或生物流体。开发工具的组合此处将促进相关的特定糖肽和糖蛋白种类的鉴定可以在预防癌症预防，进展的范围内应用的癌症样本，预后和生物标志物应用。