Chemoenzymatic methods for the detection of cell-surface glycans

检测细胞表面聚糖的化学酶法

• 批准号: 8964803
• 负责人: Peng Wu
• 金额: $ 3.35万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-15 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8964803
• 关键词: Adenocarcinoma; Adenocarcinoma In Situ; Adult; Alveolar; Antibodies; Biological Markers; Biological Models; Biopsy Specimen; Cancer Detection; Cancer Patient; Cell Communication; Cell Maturation; Cell Surface Receptors; Cell surface; Cells; Clinical; Collection; Communication; Complex; Detection; Development; Developmental Process; Diagnosis; Diagnostic; Disease Progression; Enterocytes; Enzymes; Epithelial; Epithelial Cells; Epitopes; Evolution; Excision; Freezing; Gastrointestinal Diseases; Glycobiology; Goals; Gold; Grant; Health; Histological Techniques; Histology; Human; Image; Imagery; Intestines; Label; Lasers; Lectin; Lesion; Life; Ligands; Lobectomy; Lung; Lung Adenocarcinoma; Malignant Neoplasms; Malignant neoplasm of lung; Mammalian Cell; Mammals; Mediating; Membrane; Methods; Microdissection; Monosaccharides; Normal Range; Normal tissue morphology; Oligosaccharides; Organism; Organoids; Pathologic; Pathologic Processes; Patients; Pattern; Peripheral; Physiological Processes; Play; Polysaccharides; Polysialic Acid; Process; Reagent; Research; Resected; Role; Sampling; Screening for cancer; Sensitivity and Specificity; Signal Transduction; Small Intestines; Staging; Surface; System; Techniques; Testing; Thompson-Friedenreich Antigen; Tissue Sample; Tissues; Translating; Villus; base; biological systems; biophysical chemistry; cancer therapy; cell motility; cell type; cost; disease diagnosis; glycosylation; human tissue; intestinal epithelium; lung cancer screening; minimally invasive; neoplastic; programs; receptor; self-renewal; signal processing; small molecule; stem cell differentiation; tool; tumor; tumor progression

 DESCRIPTION (provided by applicant): In mammalian organisms, nine monosaccharide building blocks are used to form cell-surface complex, higher order glycans that mediate cell-cell communication during development processes and differentiation programs. By contrast, aberrant glycosylation signals the onset of cancer. What directs pathogenic glycosylation? Is there a correlation between glycosylation patterns and cancer progression? Can we manipulate differentiation by changing the glycan coat? Despite forty years of glycobiology research, the answers to these questions remain obscure. Dynamic changes in glycosylation during many developmental processes and cancer progression are still poorly characterized. Because peripheral, higher order glycans encode information for cell-surface receptor recognition to trigger specific downstream signaling, there is an urgent and unmet need to develop methods for their detection. The long-term objective of this project is to develop chemoenzymatic methods for the specific labeling of cell-surface higher-order glycans with small-molecule biophysical probes for their characterization. To achieve this goal is to transform glycosylation enzymes into labeling reagents to target cell-surface specific glycans. Our central hypothesis is that chemoenzymatic methods for glycan detection in cells and tissue samples will have superior specificity and sensitivity than lectin- and antibody-based techniques, the current gold standard for glycan detection. In Aim 1, we will develop chemoenzymatic methods for the labeling of four sectors of glycans on the surface of live mammalian cells and transform these methods into histological techniques to label glycans in tissue sections. In Aim 2, we the small intestine developmental process. We will use the methods developed in Aim 1 to characterize the epithelial will use as a model system to study the role of glycans in a glycosylation patterns in this system. We will investigate the contribution of glycan-lectin interactions to the highly regulated cell migration and maturation in this compartment. Finally, we will apply the chemoenzymatic methods to screen cancer-associated glycans in lung resection samples from patients diagnosed with lung adenocarcinoma (Aim 3). Our goal is to investigate the potential role of these glycans in the evolution of lung cancer, and to pilot test their clinical utility forearly lung cancer detection.

 描述（由申请人提供）：在哺乳动物生物体中，9种单糖结构单元用于形成细胞表面复合物，即在发育过程和分化程序中介导细胞间通讯的高阶聚糖。相比之下，异常糖基化是癌症发作的信号。是什么引导致病性糖基化？糖基化模式与癌症进展之间是否存在相关性？我们可以通过改变聚糖涂层来操纵分化吗？尽管糖生物学研究已经进行了40年，但这些问题的答案仍然模糊不清。在许多发育过程和癌症进展中糖基化的动态变化仍然很难表征。由于外周高阶聚糖编码用于细胞表面受体识别的信息以触发特异性下游信号传导，因此迫切需要开发用于其检测的方法。 该项目的长期目标是开发化学酶法，用于用小分子生物物理探针特异性标记细胞表面高阶聚糖，以进行表征。为了实现这一目标，将糖基化酶转化为靶向细胞表面特异性聚糖的标记试剂。我们的中心假设是，用于细胞和组织样品中聚糖检测的化学酶法将具有上级基于凝集素和抗体的技术的特异性和灵敏度，这是目前聚糖检测的金标准。 在目标1中，我们将开发用于标记活哺乳动物细胞表面上的四个聚糖部门的化学酶方法，并将这些方法转化为组织学技术来标记组织切片中的聚糖。目的2：研究小肠的发育过程.我们将使用目标1中开发的方法来表征上皮细胞，将其用作模型系统来研究聚糖在该系统中糖基化模式中的作用。我们将调查的贡献聚糖凝集素相互作用的高度调节的细胞迁移和成熟在这个隔间。最后，我们将应用化学酶方法筛选诊断为肺腺癌患者的肺切除样本中的癌症相关聚糖（目标3）。我们的目标是研究这些聚糖在肺癌演变中的潜在作用，并初步测试它们在肺癌早期检测中的临床应用。