Novel synthetic tools for mucin glycobiology

粘蛋白糖生物学的新型合成工具

基本信息

批准号：
8692104
负责人：
MARE CUDIC
金额：
$ 18.56万
依托单位：
FLORIDA ATLANTIC UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-15 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8692104
关键词：
Acetylgalactosamine Adhesions Affect Affinity Allosteric Site Amino Acids Avidity Basic Science Binding Binding Proteins Biological Assay Calorimetry Cancer Vaccines Cancer cell line Carbohydrates Cell Communication Cell Proliferation Cell physiology Cell surface Cells Deposition Development Disseminated Malignant Neoplasm Epithelium Epitopes Evaluation Galactose Galactose Binding Lectin Galactosides Galectin 3 Glycobiology Glycopeptides Goals Immunity In Vitro Individual Inflammation Lead Lectin Libraries Ligands Link Malignant - descriptor Malignant Neoplasms Mediator of activation protein Membrane Modeling Molecular Mucin 1 protein Mucins National Research Council Neoplasm Metastasis Pattern Peptides Polysaccharides Premalignant Process Protein-Carbohydrate Interaction Randomized Reporting Research Proposals Role S Phase Scanning Sialic Acids Site Solid Solutions Specificity Structure Structure-Activity Relationship Technology Testing Therapeutic Agents Thompson-Friedenreich Antigen Titrations Tumor Cell Invasion Tumor-Associated Carbohydrate Antigens Vertebral column anti-cancer therapeutic base cancer cell carbohydrate binding protein clinical application combinatorial density design glycosylation immunoregulation inhibitor/antagonist insight metastatic process mouse model neoplastic cell novel novel diagnostics novel therapeutics preclinical evaluation public health relevance receptor receptor binding scaffold small molecule sugar tool tumor tumor progression

项目摘要

DESCRIPTION (provided by applicant): Aberrant cell surface glycosylation has emerged as a new hallmark of cancer. The increased expression and altered density of shorter glycoforms of mucin 1 (MUC1) are commonly observed changes in malignant and premalignant epithelia. Whereas the functional role of O-linked N-acetylgalactosamine (Tn), sialic acid capped Tn (sTn) remains unclear, Thomsen-Friedenreich (TF) antigen has been shown to be actively involved in tumor metastasis, promoting several key cell-cell interactions via association with the endogenous ¿- galactoside-specific lectin, galectin-3 (gal-3). In turn, gal-3 promotes cancer cell invasion and metastasis. Since the interaction between TF antigen and gal-3 potentially represents an important early step in heterotypic cancer-endothelial adhesion and the formation of intravascular metastatic deposits, a thorough understanding of this process at the molecular level is essential for it to be used for biomedical applications. We hypothesize that glycan presentation and the density patterns of O-linked glycans have a major impact on recognition of TF antigen of cancer-associated MUC1 by gal-3. Our recent breakthrough findings have shown that the presentation of the glycan ligand by the natural peptide scaffold is highly relevant for gal-3 binding and provides opportunities for allosteric site targeting in the design of gal-3 inhibitors. Within this proposal, we will prepare a MUC1-derived glycopeptide positional scanning combinatorial library displaying native-like heterogeneous and aberrant tumor-associated O-glycan epitopes (Aim 1). Synthesized glycopeptide libraries will be screened for gal-3 binding using a novel bead-based proximity assay based on AlphaScreen technology. The binding affinities and selectivity of the identified individual glycopeptides for gal-3 and a panelof galectins (gal-1, gal-4, gal-7, and gal-3 CRD domain) will be determined by ITC and in a more physiologically relevant setting, using cancer cell lines representing the natural complexity of glycan chains (Aim 2). MUC1-based glycopeptides that show good potency and selectivity for gal-3 will be probed for tumor cell functions relevant to tumor invasion and metastasis (Aim 3). These studies will provide insights into the functional significance of the aberrant MUC1 glycosylation by defining the role and specificity of MUC1-gal-3 interactions in cancer progression and metastasis formation. Our long-term goals are to study the specificity of MUC1-gal-3 interactions in relation to other lectin-like receptors involved in recognition and binding o tumor- associated forms of MUC1, assess other functions of MUC1/gal-3 interactions such as their role in immune modulation, and to use identified structures in design and development of small molecule-based selective and potent gal-3 inhibitors with desirable pharmacological profiles for preclinical evaluation of a novel anti-cancer therapeutic strategy.

描述（由申请人证明）：异常的细胞表面糖基化已成为粘蛋白1（MUC1）较短的糖型的新标志。唾液酸上限TN（STN）仍然是叔叔，Thomsen-Friedenreich（TF）抗原已被证明是要与肿瘤转移相关，从 - 半乳糖苷特异性凝集素，galectin-3（GAL-3）。沉积物，对分子水平的这一过程的透彻理解对于生物医学应用至关重要。 GAL-3的MUC1。与肿瘤相关的O-Glycan表位（AIM 1），使用新型的珠3型糖肽文库使用新型的珠子质量组装和Alphascreen技术进行筛选。甘染蛋白（GAL-1，GAL-4，GAL-7和GAL-3 CRD结构域）将由MUC1的基于MUC1的糖糖替代确定良好的效力和GAL-3的选择性，以探测与肿瘤功能相关的肿瘤细胞功能和转移（目标3）。 /GAL-3相互作用，例如它们在免疫模块化中的作用，以及与B le药理学概况的基于小分子的选择性和有效的GAL-3抑制剂在设计和开发中的结构，用于临床前评估