Synthetic Mucins for Structural and Compositional Studies of Mucus Gels

用于粘液凝胶结构和成分研究的合成粘蛋白

基本信息

批准号：
9760806
负责人：
AUSTIN EDWARD SCHLIRF
金额：
$ 1.71万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-01 至 2019-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9760806
关键词：
Address Adhesions Affect Amino Acid Sequence Architecture Bacteria Behavior Benchmarking Binding Binding Proteins Biochemical Biochemical Genetics Biocompatible Materials Biological Biological Products Biology Biophysics Carcinoma Characteristics Charge Chemical Structure Chronic Obstructive Airway Disease Communicable Diseases Complex Cystic Fibrosis Cystine DNA Data Diffusion Disease Disulfides Electrostatics Environment Epithelial Event Eye Eye diseases Family Fertilization Gases Gel Genes Genetic Glycobiology Glycoproteins Gynecology Health Housing Human Human Biology Human body Hydration status Hydrophobic Interactions Hydrophobicity Immunity Immunology Individual Infection Ions Jellyfish Kinetics Lead Length Ligands Lipids Liquid substance Livestock Location Malignant Neoplasms Mechanics Mediating Medical Metabolic Pathway Methodology Methods Molecular Morphology Mucin 1 protein Mucin-2 Staining Method Mucins Mucous Membrane Mucous body substance N-glycylalanine Nutrient Organism Pathologic Pathway interactions Pattern Peptides Pharmaceutical Preparations Pharmacy (field)Play Polymers Polysaccharides Preparation Process Property Protein Glycosylation Proteins RNA Splicing Recombinants Reporting Reproducibility Research Rheology Role Salts Sampling Side Skin Structure Surface Tissues Transition Elements Variant Vertebral column Water Work absorption base biophysical properties body system chemical bond chemical property chemical synthesis crosslink density design experimental study eye dryness flexibility genetic manipulation glutamylalanine glycosylation host microbiome human tissue leucyl-alanine materials science microbiome mimetics molecular size monomer natural Blastocyst Implantation novel therapeutics nutrient absorption particle pathogen physical property polymerization reconstitution respiratory retinal rods self assembly small molecule success tool

项目摘要

Abstract Mucus is the primary material that mediates interactions with the outside world in organisms from humans to jellyfish. Mucus gels function as a hydrating barrier involved in events such as embryo implantation, absorption of nutrients, drugs, and pathogens, while also housing the majority of the microbiome. Despite these essential roles, mucus composition, physical properties, and biology remain poorly defined. This is because the major component, mucin glycoproteins, is innately heterogeneous and cannot be reproducibly obtained by any current methodology. This roadblock has hindered our understanding of epithelial biology across diverse fields. The overall objective of this proposal is to generate synthetic mucus as transformative materials to probe the structure and function of native mucus, and with biomedical applications treating compromised tissues. We hypothesize that synthetic multi-block glycopolypeptides can emulate natural multi-domain gel-forming mucins, but with precisely defined and tunable compositions capable of selective modulation of gel properties and bioactivity. Native mucins are a family of 20+ glycoproteins characterized by massive rod-like domain rich in glycosylated-Ser/Thr, and short terminal domains that play a role in formation of cross-linked mucins bundles via Cys disulfides and hydrophobic interactions. Mucin expression and splice variation are unique to each tissue and disease, and the proteins' glycosylation patterns are the product of complex metabolic pathways controlled by >1000 genes. These pathways are poorly understood and cannot be manipulated by any current genetic or biochemical methods. Overall, biological mucins are too heterogeneous to probe many specific hypotheses. Glycopolymers have been explored as mucus-mimics, but prior examples have failed to recapitulate the chemical structures and biophysics of native mucins. During the project period, we will 1) develop tunable and reproducible synthetic mucins based on multi- block glycopolypeptides that faithfully emulate the chemical and biophysical properties of natural mucins, and 2) unravel how mucus composition (pH, ions, lipids, DNA, proteins) affects both gel physical properties and glycan-dependent bioactivity. We will precisely tune the glycan patterns by chemical synthesis and enzymatic glycosylation to prepare binding or control ligands to interact with glycan-binding proteins. These properties cannot be controlled by any other current methods. We will assemble the glycopolypeptides into gels with varied compositions inspired by analysis of native mucus, and we will benchmark our materials against commercially available mucins. We expect to provide new tools for our lab and others to study previously untestable hypotheses regarding mucosal transport and biology relevant to health and disease. Success of the proposed research is anticipated to make a transformative impact across diverse fields from materials science and glycobiology to pharmaceutics, immunology, infectious diseases, gasteroenterology, and gynecology.

抽象的粘液是介导从人类到生物体与外界相互作用的主要物质海蜇。粘液凝胶充当水合屏障，参与胚胎植入、吸收等事件营养物质、药物和病原体，同时也容纳了大部分微生物组。尽管有这些必要的作用、粘液成分、物理性质和生物学仍然不明确。这是因为主要成分粘蛋白糖蛋白本质上是异质的，不能通过任何方法重复获得当前的方法。这一障碍阻碍了我们对不同领域的上皮生物学的理解。该提案的总体目标是生成合成粘液作为转化材料来探索天然粘液的结构和功能，以及治疗受损组织的生物医学应用。我们假设合成的多嵌段糖多肽可以模拟天然的多结构域凝胶形成粘蛋白，但具有精确定义和可调的成分，能够选择性调节凝胶特性和生物活性。天然粘蛋白是一个由 20 多种糖蛋白组成的家族，其特征是大量的杆状结构域富含糖基化 Ser/Thr 和短末端结构域，在交联形成中发挥作用粘蛋白束通过半胱氨酸二硫化物和疏水相互作用。粘蛋白表达和剪接变异每种组织和疾病都是独特的，蛋白质的糖基化模式是复杂的产物代谢途径由超过 1000 个基因控制。人们对这些途径知之甚少并且无法通过任何当前的遗传或生化方法进行操纵。总体而言，生物粘蛋白太异质来探究许多特定的假设。糖聚合物已被探索作为粘液模拟物，但是先前的例子未能概括天然粘蛋白的化学结构和生物物理学。在项目期间，我们将1）开发基于多因素的可调节和可重复的合成粘蛋白阻断糖多肽，忠实地模拟天然粘蛋白的化学和生物物理特性， 2) 揭示粘液成分（pH、离子、脂质、DNA、蛋白质）如何影响凝胶物理特性和聚糖依赖性生物活性。我们将通过化学合成精确调整聚糖模式酶促糖基化以制备结合或控制配体以与聚糖结合蛋白相互作用。这些属性不能由任何其他当前方法控制。我们将糖多肽组装成受到天然粘液分析的启发，我们制作了具有不同成分的凝胶，我们将对我们的材料进行基准测试针对市售粘蛋白。我们期望为我们的实验室和其他人的研究提供新的工具关于与健康和疾病相关的粘膜运输和生物学的先前无法检验的假设。预计拟议研究的成功将对各个领域产生变革性影响材料科学和糖生物学到药剂学、免疫学、传染病、胃肠病学、和妇科。