Novel synthetic tools for mucin glycobiology

粘蛋白糖生物学的新型合成工具

• 批准号: 8927562
• 负责人: MARE CUDIC
• 金额: $ 20.58万
• 依托单位: FLORIDA ATLANTIC UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-15 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8927562
• 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; Health; 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; receptor; receptor binding; scaffold; small molecule; sugar; tool; tumor; tumor progression

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（由申请人提供）：异常细胞表面糖基化已成为癌症的新标志。mucin 1 （MUC1）的短糖型表达增加和密度改变是在恶性和癌前上皮中常见的变化。尽管O-linked n- acetyl半乳糖胺（Tn）、唾液酸帽Tn （sTn）的功能作用尚不清楚，但thomson - friedenreich （TF）抗原已被证明积极参与肿瘤转移，通过与内源性半乳糖苷特异性凝集素半乳糖素-3 （gal-3）的关联，促进了几种关键的细胞-细胞相互作用。反过来，gal-3促进癌细胞的侵袭和转移。由于TF抗原和gal-3之间的相互作用可能代表了异型癌症内皮粘附和血管内转移沉积形成的重要早期步骤，因此在分子水平上对这一过程的透彻理解对于将其用于生物医学应用至关重要。我们假设，聚糖的呈现和o链聚糖的密度模式对gal-3对癌症相关MUC1的TF抗原的识别有重要影响。我们最近的突破性发现表明，天然肽支架提供的聚糖配体与gal-3结合高度相关，并为gal-3抑制剂设计中的变构位点靶向提供了机会。在本提案中，我们将准备一个muc1衍生的糖肽定位扫描组合文库，显示天然样异质和异常肿瘤相关的o -聚糖表位（Aim 1）。合成的糖肽文库将使用基于alphasgreen技术的新型基于头的接近试验筛选gal-3结合。所鉴定的单个糖肽对gal-3和一组凝集素（gal-1、gal-4、gal-7和gal-3 CRD结构域）的结合亲和力和选择性将由ITC和在更生理相关的环境中确定，使用代表聚糖链自然复杂性的癌细胞系（目的2）。基于muc1的糖肽对gal-3表现出良好的效力和选择性，将用于研究与肿瘤侵袭和转移相关的肿瘤细胞功能（目的3）。这些研究将通过定义MUC1-gal-3相互作用在癌症进展和转移形成中的作用和特异性，深入了解MUC1异常糖基化的功能意义。我们的长期目标是研究MUC1-gal-3相互作用与其他凝集素样受体的特异性，这些受体参与MUC1肿瘤相关形式的识别和结合，评估MUC1/gal-3相互作用的其他功能，例如它们在免疫调节中的作用。并利用已确定的结构设计和开发基于小分子的选择性和有效的gal-3抑制剂，这些抑制剂具有理想的药理学特征，用于新型抗癌治疗策略的临床前评估。

项目成果

Targeting cancer-specific glycans by cyclic peptide lectinomimics.

通过环肽凝集素靶向癌症特异性聚糖。

• DOI: 10.1007/s00726-017-2485-3
• 发表时间: 2017
• 期刊: Amino acids
• 影响因子: 3.5
• 作者: Rodriguez,MariaC;Yongye,AustinB;Cudic,Mihael;MartinezMayorga,Karina;Liu,Enbo;Mueller,BarbaraM;Ainsley,Jon;Karabencheva-Christova,Tatyana;Christov,ChristoZ;Cudic,Mare;Cudic,Predrag
• 通讯作者: Cudic,Predrag

How altering the modular architecture affects aspects of lectin activity: case study on human galectin-1.

• DOI: 10.1093/glycob/cwz034
• 发表时间: 2019-05
• 期刊: Glycobiology
• 影响因子: 4.3
• 作者: Tanja J. Kutzner;A. Gabba;Forrest G. FitzGerald;N. Shilova;Gabriel García Caballero;Anna‐Kristin Ludwig;J. C. Manning;C. Knospe;H. Kaltner;F. Sinowatz;P. Murphy;M. Cudic;N. Bovin;H. Gabius

Thermodynamic Switch in Binding of Adhesion/Growth Regulatory Human Galectin-3 to Tumor-Associated TF Antigen (CD176) and MUC1 Glycopeptides.

粘附/生长调节人半乳糖凝集素 3 与肿瘤相关 TF 抗原 (CD176) 和 MUC1 糖肽结合的热力学转换。

• DOI: 10.1021/acs.biochem.5b00555
• 发表时间: 2015
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Rodriguez,MariaC;Yegorova,Svetlana;Pitteloud,Jean-Philippe;Chavaroche,AnaisE;André,Sabine;Ardá,Ana;Minond,Dimitriy;Jiménez-Barbero,Jesús;Gabius,Hans-Joachim;Cudic,Mare
• 通讯作者: Cudic,Mare