"A molecular approach toward elucidating the role of the mucin glycocalyx in canc

“阐明粘蛋白糖萼在癌症中作用的分子方法

• 批准号: 9143863
• 负责人: Jessica Kramer
• 金额: $ 2.83万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-02-01 至 2017-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9143863
• 关键词: Address; Adhesions; Affect; Aldehydes; Apoptotic; Biochemical; Biological; Biological Markers; Biology; Breast; Breast Epithelial Cells; Cancer Vaccines; Cell Adhesion; Cell Adhesion Molecules; Cell Survival; Cell surface; Cells; Cellular biology; Chimera organism; Chimeric Proteins; Collection; Colon Carcinoma; Complex; Detection; Development; Embryonic Development; Engineering; Epidermal Growth Factor Receptor; Epithelial; Epitopes; Extracellular Domain; Extracellular Matrix; Fluorescent Probes; Focal Adhesion Kinase 1; Focal Adhesions; Galactose Binding Lectin; Genes; Genetic; Genetic Polymorphism; Glycobiology; Glycocalyx; Glycoproteins; Goals; Growth; Growth Factor; Health; Human; Immune Cell Activation; Immune system; In Vitro; Integrins; Kinetics; Lead; Length; Libraries; Life; Light; Lipids; MCF10A cells; Malignant Neoplasms; Mechanics; Membrane; Membrane Glycoproteins; Metabolic Pathway; Methods; Microscopic; Molecular; Mucin-1 Staining Method; Mucins; Nature; Neoplasm Metastasis; Non-Malignant; Ovarian Carcinoma; Pancreatic carcinoma; Pattern; Phase; Play; Polymer Chemistry; Polysaccharides; Process; Property; Protein Engineering; Regulation; Research; Role; Signal Transduction; Structure; Supplementation; Surface; Suspension substance; Suspensions; Techniques; Tissues; cancer cell; cell motility; crosslink; density; extracellular; glycosylation; insight; interdisciplinary approach; lung Carcinoma; migration; mimetics; novel therapeutic intervention; overexpression; tumor progression

DESCRIPTION (provided by applicant): Cell-surface glycans play essential roles in diverse cell surface interactions, yet glycobiology studies are complicated by the varied and dynamic nature of glycan presentation. New techniques to precisely modify and engineer glycans on the cell surface are of paramount importance for crucial insights into processes such as immune cell activation, embryonic development, and cancer progression. An illustrative example of the glycobiology challenge is the membrane associated mucin glycoprotein MUC1. MUC1 is highly over-expressed and aberrantly glycosylated on ca. 90% of breast, ovarian, lung, colon, and pancreatic carcinomas, is a biomarker for detection of epithelial tissue derived cancers, and is a target for cancer vaccines. MUC1 studies have been complicated by its sheer size (150-300 kDa), polymorphism, variable glycosylation patterns, and because both the cytosolic and extracellular domains can participate in signaling that affects cell survival and migration. There is a widespread assumption that cancer-associated mucins sterically block access of cell surface adhesion molecules to the extracellular matrix and shield cancer cells from the immune system; yet molecular evidence is lacking and recent evidence has suggested more sophisticated roles. The proposed research seeks to understand the functional significance of cancer-associated mucin overexpression by using a new method of cell surface engineering that will enable correlation of subtle changes in glycosylation with biochemical actions. A library of synthetic glycopolymer mimics of MUC1's extracellular glycodomain will be prepared, attached to membrane anchoring lipids or engineered MUC1 protein chimeras, and displayed on live cells. Using microscopic and biochemical methods, I will investigate how changes in the mucin glycocalyx influence mechanical regulation of integrin clustering, focal adhesion formation, and signaling that has downstream effects on cellular adhesion, survival, and migration. This research could lead to a new understanding of how the glycocalyx affects extracellular matrix interactions pertinent to cancer progression. Additionally, development of our cell surface engineering method has strong potential for broad applications in studying diverse cell surface interactions. Overall, this interdisciplinary approach will combine techniques in polymer chemistry and cell biology to answer important questions about cancer progression that cannot be undertaken by biological methods alone.

描述（由申请方提供）：细胞表面聚糖在多种细胞表面相互作用中发挥重要作用，但聚糖呈递的多样性和动态性使糖生物学研究复杂化。精确修饰和改造细胞表面聚糖的新技术对于深入了解免疫细胞活化、胚胎发育和癌症进展等过程至关重要。糖生物学挑战的说明性实例是膜相关粘蛋白糖蛋白MUC 1。MUC 1在CA上高度过表达并异常糖基化。90%的乳腺癌、卵巢癌、肺癌、结肠癌和胰腺癌，是用于检测上皮组织来源的癌症的生物标志物，并且是癌症疫苗的靶标。MUC 1的研究因其庞大的尺寸（150-300 kDa）、多态性、可变的糖基化模式而变得复杂，并且因为胞质和胞外结构域都可以参与影响细胞存活和迁移的信号传导。有一个广泛的假设，即癌症相关粘蛋白空间阻断细胞表面粘附分子的细胞外基质的访问和屏蔽癌细胞从免疫系统，但缺乏分子证据，最近的证据表明更复杂的作用。拟议的研究旨在通过使用一种新的细胞表面工程方法来理解癌症相关粘蛋白过表达的功能意义，该方法将使糖基化的细微变化与生化作用相关。图书馆 将制备MUC 1胞外糖结构域的合成糖聚合物模拟物，连接到膜锚定脂质或工程化MUC 1蛋白嵌合体上，并在活细胞上展示。使用显微镜和生物化学方法，我将研究粘蛋白糖萼的变化如何影响整合素聚集，粘着斑形成和信号传导的机械调节，对细胞粘附，存活和迁移具有下游效应。这项研究可能会导致对糖萼如何影响与癌症进展相关的细胞外基质相互作用的新理解。此外，我们的细胞表面工程方法的发展在研究不同的细胞表面相互作用具有广泛的应用潜力。总的来说，这种跨学科的方法将联合收割机技术 在聚合物化学和细胞生物学中，回答了关于癌症进展的重要问题，这些问题不能单独通过生物学方法来解决。