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

• 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
• 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

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.

摘要 粘液是介导从人类到人类的生物体与外部世界相互作用的主要物质， 水母粘液凝胶作为一种水合屏障，参与胚胎植入、吸收等过程。 营养物质，药物和病原体，同时也容纳了大多数微生物。尽管这些基本 作用、粘液成分、物理性质和生物学仍然不明确。这是因为少校 组分粘蛋白糖蛋白是先天异质的，并且不能通过任何方法可重复地获得。 目前的方法。这个障碍阻碍了我们对不同领域上皮生物学的理解。 该提案的总体目标是产生合成粘液作为转化材料，以探测 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，但具有能够选择性调节凝胶性质的精确定义和可调的组成 和生物活性。天然粘蛋白是一个由20多个糖蛋白组成的家族，其特征在于具有大量的棒状结构域 富含糖基化的Ser/Thr和短末端结构域，其在形成交联的 粘蛋白束通过Cys二硫化物和疏水相互作用。粘蛋白表达和剪接变异 每个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。 由超过1000个基因控制的代谢途径。这些途径知之甚少， 被任何现有的遗传或生化方法操纵。总的来说，生物粘蛋白也 异质性，以探讨许多具体的假设。糖共聚物已被探索为粘液模拟物，但 先前的实施例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）开发可调谐和可重复的合成粘蛋白， 嵌段糖多肽忠实地模拟天然粘蛋白的化学和生物物理性质， 以及2）阐明粘液成分（pH、离子、脂质、DNA、蛋白质）如何影响凝胶物理性质 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， 酶促糖基化以制备与聚糖结合蛋白相互作用的结合或对照配体。这些 属性不能由任何其他当前方法控制。我们将把糖多肽组装成 凝胶与不同的组成灵感来自分析天然粘液，我们将基准我们的材料 针对市售粘蛋白。我们希望为我们的实验室和其他人提供新的工具来研究 关于粘膜运输和与健康和疾病相关的生物学的先前不可检验的假设。 预计拟议研究的成功将对不同领域产生变革性影响， 材料科学和糖生物学到制药学，免疫学，传染病学，胃肠病学， 和妇科