Development of Mucin-mimetics to Study the Role of the Glycocalyx in Oncogenesis

开发粘蛋白模拟物以研究糖萼在肿瘤发生中的作用

• 批准号: 9031563
• 负责人: Elliot Carter Woods
• 金额: $ 3.73万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-05 至 2017-03-04
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9031563
• 关键词: Achievement; Adhesions; Affect; Alkanes; Apoptosis; Automobile Driving; Binding; Biochemical; Biological; Biological Assay; Biological Process; Carbohydrate Chemistry; Carcinoma; Cell Survival; Cell membrane; Cell physiology; Cell surface; Cells; Charge; Chemicals; Complex; Computer Analysis; Crowding; Cytoskeleton; Data; Development; Elements; Endocytosis; Engineering; Epithelium; Event; Exhibits; Extracellular Domain; Focal Adhesion Kinase 1; Focal Adhesions; Genetic; Genetic Techniques; Glycocalyx; Growth; Health; Hour; Integrin Binding; Integrins; Ketones; Kinetics; Length; Ligands; Link; Lipids; Malignant - descriptor; Malignant Neoplasms; Measures; Mechanics; Mediating; Membrane; Membrane Glycoproteins; Membrane Proteins; Methods; Microscopy; Modeling; Molecular Biology Techniques; Mucin 1 protein; Mucins; Nature; Neoplasm Metastasis; Non-Malignant; Outcome; Phenotype; Phosphorylation; Play; Polymer Chemistry; Polymers; Polysaccharides; Recycling; Role; Signal Transduction; Structure; Structure-Activity Relationship; Tail; Technology; Testing; Time; Up-Regulation; Variant; Vertebral column; Western Blotting; Work; base; cell growth; density; experience; improved; in vivo; malignant breast neoplasm; malignant phenotype; migration; mimetics; next generation; overexpression; physical property; polyacrylamide gels; research study; residence; sugar; tool; trait; tumor; tumor progression; tumorigenesis

 DESCRIPTION (provided by applicant): Mucins are upregulated in more than 90% of breast cancers and many other carcinomas, and their link to oncogenesis is clear. Still, the role of their large extracellular domains-a major structural component of the glycocalyx-in this oncogenesis has yet to be elucidated. Because it is known that integrin-mediated adhesion can play a critical role in such diverse cellular processes as differentiation, proliferation, migration and apoptosis, aberrant integrin function has been linked to oncogenesis in myriad ways. Preliminary studies suggest that mucins could play a role in boosting the pro-malignant functions of integrin signaling by changing the physical properties of the glycocalyx. Mucins are heavily O-glycosylated membrane-associated proteins involved in the protection of epithelia from luminal insults. Because they often consist of greater than 50% glycan by mass, their structures are not easily manipulated using standard molecular biology techniques. An approach to studying these molecules relying on synthetic mucin-mimetic glycopolymers has proven a tractable method to overcome this obstacle. Well-defined, polymeric alkane backbones with pendant ketones are easily decorated with aminooxy-glycans. One end of the polymer contains a probe for microscopy and the other a hydrophobic tail for spontaneous insertion into cell membranes. In this proposal we will test the hypothesis that the glycocalyx affects integrin-mediated adhesion in a structure dependent manner, and thus drives a metastatic phenotype, by utilizing these mucin-mimetics. In Specific Aim 1, mucin-mimetic glycopolymers will be improved by increasing their ability to reside on cell surfaces for longer periods of time by chemically modifying their hydrophobic anchors. Increasing cell-surface half-lives will vastly improve these polymers as tools with which to study the glycocalyx and its components. In Specific Aim 2, structural variation of the glycocalyx will be studied using these tools and its effect on integrin- driven metastatic phenotype will be characterized. If confirmed, this hypothesis would present a paradigm shift for the role of the mucins, and the glycocalyx, in oncogenesis.

 描述(申请人提供)：粘蛋白在超过90%的乳腺癌和许多其他癌症中表达上调，它们与肿瘤发生的联系是明确的。尽管如此，他们的角色 大的胞外区--糖萼的主要结构成分--在这一肿瘤发生中的作用尚未阐明。因为已知整合素介导的黏附在分化、增殖、迁移和凋亡等多种细胞过程中发挥关键作用， 整合素功能的异常与肿瘤的发生有多种联系。初步研究表明，粘蛋白可以通过改变糖萼的物理性质，在增强整合素信号的促恶性功能方面发挥作用。粘蛋白是一种高度O-糖基化的膜相关蛋白，参与保护上皮细胞免受管腔损伤。因为它们通常由质量超过50%的多糖组成，所以它们的结构不容易使用标准的分子生物学技术来操纵。一种依靠合成的粘蛋白模拟糖共聚物来研究这些分子的方法已经证明是克服这一障碍的一种容易处理的方法。定义明确的、带有悬垂酮的聚合烷烃主链很容易被氨氧基-葡聚糖装饰。聚合物的一端包含用于显微镜观察的探针，另一端包含用于自发插入细胞膜的疏水尾巴。在这项建议中，我们将通过利用这些粘蛋白模拟物来检验这一假设，即糖萼以一种结构依赖的方式影响整合素介导的黏附，从而驱动转移表型。在具体目标1中，拟粘蛋白糖共聚物将通过对其疏水锚点进行化学修饰来提高其在细胞表面停留更长时间的能力。增加细胞表面的半衰期将极大地改善这些聚合物作为研究糖萼及其成分的工具。在特定的目标2中，将使用这些工具研究糖萼的结构变异，并将表征其对整合素驱动的转移表型的影响。如果得到证实，这一假说将代表着粘蛋白和糖萼在肿瘤发生中的作用的范式转变。

项目成果

Targeted glycan degradation potentiates the anticancer immune response in vivo.

• DOI: 10.1038/s41589-020-0622-x
• 发表时间: 2020-12
• 期刊: Nature chemical biology
• 影响因子: 14.8
• 作者: Gray MA;Stanczak MA;Mantuano NR;Xiao H;Pijnenborg JFA;Malaker SA;Miller CL;Weidenbacher PA;Tanzo JT;Ahn G;Woods EC;Läubli H;Bertozzi CR
• 通讯作者: Bertozzi CR