"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

基本信息

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
项目类别：
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

项目摘要

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（由申请人提供）：细胞表面聚糖在各种细胞表面相互作用中起着重要的作用，但是糖生物学研究因聚糖表现的多样性和动态性质而变得复杂。精确修改细胞表面的Glycans的新技术对于对免疫细胞激活，胚胎发育和癌症进展等过程的关键见解至关重要。糖生物学挑战的一个说明性例子是膜相关的粘蛋白糖蛋白MUC1。 MUC1在CA上高表达和异常糖基化。 90％的乳腺癌，卵巢，肺，结肠和胰腺癌是检测上皮组织衍生癌症的生物标志物，并且是癌症疫苗的靶标。 MUC1研究的大小（150-300 kDa），多态性，可变糖基化模式以及细胞质和细胞外域都可以参与影响细胞存活和迁移的信号传导，这使MUC1研究变得复杂。有一个广泛的假设是，与癌症相关的粘蛋白在肠外基质中阻止细胞表面粘附分子的访问，并避免免疫系统中的癌细胞屏蔽癌细胞。然而，缺乏分子证据，最近的证据表明角色更复杂。拟议的研究试图通过使用一种新的细胞表面工程方法来理解与癌症相关的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。图书馆将准备MUC1细胞外糖结域的合成糖聚合物模仿，并附着在膜锚定脂质或工程的MUC1蛋白嵌合体上，并显示在活细胞上。使用显微镜和生化方法，我将研究粘蛋白糖脂的变化如何影响整联蛋白聚类，局灶性粘附形成以及对细胞粘附，生存和迁移的下游影响的信号传导的机械调节。这项研究可能会导致对糖脂的新了解如何影响与癌症进展有关的细胞外基质相互作用。此外，我们的细胞表面工程方法的开发在研究各种细胞表面相互作用方面具有强大的应用。总体而言，这种跨学科方法将结合技术在聚合物化学和细胞生物学中，回答有关癌症进展的重要问题，仅生物学方法就无法进行。