Chemical Cell Surface Engineering

化学细胞表面工程

基本信息

批准号：
9130185
负责人：
Carolyn Bertozzi
金额：
$ 42.84万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-08-01 至 2018-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9130185
关键词：
Affect Allografting Behavior Binding Biological Biological Process Biology Breast Epithelial Cells Cancer Biology Cancerous Cell Adhesion Cell Surface Proteins Cell Survival Cell Transplants Cell membrane Cell surface Cells Chemicals Chemistry Collection Complex Cultured Cells Diffusion Engineering Epidermal Growth Factor Receptor Extracellular Matrix Galactose Binding Lectin Glycobiology Glycocalyx Glycoconjugates Graft Rejection Grant Health Human Immune response Immune system In Vitro Integrins Islet Cell Islets of Langerhans Transplantation Kinetics Life Ligands Lipids Malignant Neoplasms Mediating Metastatic Neoplasm to the Lung Methods Modeling Molecular Mucin 1 protein Mucins Natural Killer Cells Neoplasm Metastasis Nucleic Acids Nylons Organ Transplantation Plant Roots Polysaccharides Predisposition Proteins Role Signal Transduction Structure Suspension substance Suspensions Tail Technology Time Vertebral column Work Xenograft procedure allograft rejection base cancer cell cancer therapy cell killing design extracellular in vitro Assay in vivo insight islet allograft killings overexpression physical property prevent receptor residence sialic acid binding Ig-like lectin sugar tool trafficking tumor

项目摘要

DESCRIPTION (provided by applicant): Glycans attached to cell surface proteins and lipids mediate interactions with receptors on other cells, in the extracellular matrix, or within the same cell membrane. Collectively, cell-surface glycans constitute a "glycocalyx" with bulk physical properties that can also influence extracellular interactions. The biological functions of natural glycoconjugates are difficult to study due to a dearth of experimental tools. Unlike proteins and nucleic acids, glycan structures are impossible to alter with molecular precision using biological methods. The broad objective of this project is to develop chemical approaches that enable fundamental studies of cell surface glycobiology. In the last granting period, we designed synthetic glycopolymers that emulate the structures of mucin glycodomains and can be anchored to cell membranes through a lipid tail. We demonstrated that these fully synthetic materials, whose structures can be modulated with precision, can be introduced onto live cells where they acquire biological activity. In collaborative work with Prof. Valerie Weaver (UCSF) we used the glycopolymers as models of MUC1, a cell-surface mucin that is overexpressed on many cancers. Mammary epithelial cells remodeled with the glycopolymers underwent changes in integrin clustering and extracellular matrix binding that were similar to the effects of MUC1 overexpression. These results validated synthetic glycopolymers as functionally relevant tools for unraveling the biology of the cancer glycocalyx. In the next granting period, we will build upon our work with synthetic glycopolymers with three Specific Aims. In Aim 1, we will build a collection of glycopolymers possessing varied glycan structures, more biologically authentic backbones, and tunable plasma membrane residence times. In Aim 2, we will continue our collaborative work with Prof. Weaver's group, using the glycopolymers from Aim 1 to ascertain the roles of mucin glycodomains in regulating cell adhesion, survival and proliferation in vitro, and metastasis in vivo. In Aim 3 we will use glycopolymers to probe the involvement of SigLecs in NK cell-mediated transplant rejection; we also seek to develop immunoprotective materials for islet cell transplants based on our findings.

描述（由申请人提供）：连接到细胞表面蛋白和脂质的聚糖介导与其他细胞上的受体，细胞外基质或在同一细胞中的相互作用细胞膜。总体而言，细胞表面聚糖构成具有散装物理特性的“糖椰子”，也可以影响细胞外相互作用。由于缺乏实验工具，因此很难研究天然糖缀合物的生物学功能。与蛋白质和核酸不同，使用生物学方法可以通过分子精度改变聚糖结构。该项目的广泛目的是开发化学方法，以实现细胞表面糖生物学的基本研究。在最后一个授予期间，我们设计了模仿粘蛋白糖结域结构的合成糖聚合物，并可以通过脂质尾部固定在细胞膜上。我们证明了这些完全合成的材料可以用精度调节其结构，可以将其引入生物活性的活细胞中。在与Valerie Weaver教授（UCSF）的合作工作中，我们使用了糖聚合物作为MUC1的模型，MUC1是一种细胞表面粘蛋白，在许多癌症上都过表达。用糖聚合物重塑的乳腺上皮细胞经历了整联蛋白聚类和细胞外基质结合的变化，与MUC1过表达的作用相似。这些结果验证了合成糖聚合物作为阐明癌症糖蛋白生物学的功能相关工具。在下一个授予期间，我们将以三个特定目标的合成糖聚合物的工作为基础。在AIM 1中，我们将建立一系列具有多种聚糖结构，更真实的骨架和可调的质膜停留时间的聚糖聚合物的集合。在AIM 2中，我们将使用AIM 1的糖聚合物继续与Weaver教授的合作合作，以确定粘蛋白糖酶在调节细胞粘附，体外的生存和增殖以及体内转移中的作用。在AIM 3中，我们将使用糖聚合物探测Siglecs在NK细胞介导的移植排斥反应中的参与。我们还试图根据我们的发现开发用于胰岛细胞移植的免疫保护材料。

项目成果

期刊论文数量（47）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A 96-well dot-blot assay for carbohydrate sulfotransferases.

碳水化合物磺基转移酶的 96 孔斑点印迹测定。

DOI：
10.1016/s0003-2697(02)00060-x
发表时间：
2002
期刊：
Analytical biochemistry
影响因子：
2.9
作者：
Verdugo,DawnE;Bertozzi,CarolynR
通讯作者：
Bertozzi,CarolynR

The glycosylphosphatidylinositol anchor: a complex membrane-anchoring structure for proteins.

DOI：
10.1021/bi8006324
发表时间：
2008-07-08
期刊：
BIOCHEMISTRY
影响因子：
2.9
作者：
Paulick, Margot G.;Bertozzi, Carolyn R.
通讯作者：
Bertozzi, Carolyn R.

Investigating cell surface galectin-mediated cross-linking on glycoengineered cells.

DOI：
10.1021/ja301694s
发表时间：
2012-06-13
期刊：
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
影响因子：
15
作者：
Belardi, Brian;O'Donoghue, Geoff P.;Smith, Adam W.;Groves, Jay T.;Bertozzi, Carolyn R.
通讯作者：
Bertozzi, Carolyn R.

Synthesis of heterobifunctional protein fusions using copper-free click chemistry and the aldehyde tag.

DOI：
10.1002/anie.201108130
发表时间：
2012-04-23
期刊：
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
影响因子：
16.6
作者：
Hudak, Jason E.;Barfield, Robyn M.;de Hart, Gregory W.;Grob, Patricia;Nogales, Eva;Bertozzi, Carolyn R.;Rabuka, David
通讯作者：
Rabuka, David

Cu-free click cycloaddition reactions in chemical biology.