Chemoenzymatic methods for the detection of cell-surface glycans

检测细胞表面聚糖的化学酶法

基本信息

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
项目类别：
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

项目摘要

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，九种单糖结构单元用于形成细胞表面复合物、在发育过程和分化程序期间介导细胞间通讯的高级聚糖。相比之下，异常糖基化标志着癌症的发生。是什么指导致病性糖基化？糖基化模式与癌症进展之间是否存在相关性？尽管经过四十年的糖生物学研究，我们能否通过改变聚糖外壳来控制分化？这些问题仍然不清楚。许多发育过程和癌症进展过程中糖基化的动态变化仍然知之甚少，因为外周的高级聚糖编码细胞表面受体识别的信息以触发特定的下游信号传导，因此迫切需要解决这一问题。该项目的长期目标是开发用小分子生物物理探针特异性标记细胞表面高阶聚糖的酶法，以实现其表征。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方法将比基于凝集素和抗体的技术（当前聚糖检测的黄金标准）具有更高的特异性和灵敏度。目标 1，我们将开发用于标记活哺乳动物细胞表面的四个聚糖部分的化学酶方法，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。我们将使用目标 1 中开发的方法来表征上皮细胞，并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。最后，我们将应用化学酶法筛选肺腺癌患者肺切除样本中的癌症相关聚糖（目标 3）。目的是研究这些聚糖在肺癌进化中的潜在作用，并初步测试它们在早期肺癌检测中的临床实用性。