Synthetic mucins in epithelial models to probe virus-mucin interactions

上皮模型中的合成粘蛋白用于探测病毒-粘蛋白相互作用

• 批准号: 10655654
• 负责人: Jessica Kramer
• 金额: $ 38.5万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10655654
• 关键词: Adhesions; Amino Acids; Antiviral Agents; Binding; Biology; Cell surface; Cells; Complex; Development; Diet; Diffusion; Disease; Drug Delivery Systems; Engineering; Epithelial Cells; Epithelium; Evolution; Future; Glycocalyx; Glycoproteins; Health; Heterogeneity; Host Defense; Human; Hydration status; Immunity; Infection; Jellyfish; Knowledge; Life; Life Cycle Stages; Lubrication; Malignant Neoplasms; Mentors; Modeling; Molecular; Molecular Weight; Mucins; Mucous body substance; Outcome; Pathway interactions; Pattern; Peptides; Play; Polymers; Polysaccharides; Prevention strategy; Proteins; Role; Sialic Acids; Specificity; Structure; Tissues; Underrepresented Populations; Vertebral column; Viral; Viral Genes; Virus; Virus Diseases; biophysical properties; chemical property; citizen science; disease transmission; equity, diversity, and inclusion; glycosylation; improved; novel therapeutics; nutrient absorption; pathogen; polymerization; polypeptide; preference; response; sugar; surface coating; tissue tropism

PROJECT SUMMARY Mucin glycoproteins are the essential component of mucus and the epithelial cellular glycocalyx. Mucins are essential for life in creatures from jellyfish to humans and play roles in hydration, lubrication, nutrient absorption, and host defense against pathogens. Mucin glycosylation is regulated by complex enzymatic pathways subject to flux, resulting in heterogeneous and variable glycan patterns that vary between tissues and species, and that evolve in response to diet and disease. The Kramer Lab is developing synthetic mucins, or synMUCs, that harness the chemical and biophysical properties of native mucins but have molecularly tunable structures. Polymerization of glycosylated amino acid N-carboxyanhydrides affords high molecular weight polypeptides with the native peptide and glycan linkages. Compared to short peptides, polysaccharides, or traditional polymers bearing attached sugars, synMUCs are the most authentic mucin mimics to date. The synMUCs will be applied in engineered models of the glycocalyx and secreted mucus. These models will find broad future application in studies of epithelial biology with application in cancer, drug delivery, immunity, and infection. Since mucins are on the front lines of cellular defense, diverse viruses have evolved strategies to adhere to their glycans, alter them, and even use them to enter host cells for replication. Virus-mucin binding can have outcomes on viral diffusion, tissue specificity, and replication but molecular details are lacking due to mucin heterogeneity. We will chemoenzymatically modify our synMUCs to display virus-binding sialic acid glycans. Viral binding preferences for various sialic acid structures in different densities and from varied peptide backbone compositions will be defined. The sialic-acid-bearing-synMUCs will be utilized to probe how mucins in the glycocalyx vs mucus regulate adhesion, cell entry and replication, tissue tropism and viral gene evolution. This knowledge will shed light on fundamental aspects of the viral life cycle and may assist in improving human health though development of new antiviral therapeutics and disease transmission prevention strategies. Additionally, scientific citizenship and mentoring are a priority and active involvement in supporting equity, diversity and inclusion of underrepresented groups in STEM will be a focus for the duration of the project and beyond.

项目概要 粘蛋白糖蛋白是粘液和上皮细胞糖萼的重要组成部分。粘蛋白是 从水母到人类，对生物的生命至关重要，在水合作用、润滑、营养吸收、 和宿主对病原体的防御。粘蛋白糖基化受复杂酶途径调节 流动，导致不同组织和物种之间存在异质和可变的聚糖模式，并且 进化以应对饮食和疾病。 Kramer 实验室正在开发合成粘蛋白（synMUC）， 利用天然粘蛋白的化学和生物物理特性，但具有分子可调结构。 糖基化氨基酸 N-羧酸酐的聚合可提供高分子量多肽 天然肽和聚糖连接。与短肽、多糖或传统聚合物相比 synMUC 带有附着的糖，是迄今为止最真实的粘蛋白模拟物。 synMUC 将被应用 在糖萼和分泌粘液的工程模型中。这些模型将在未来得到广泛的应用 上皮生物学研究及其在癌症、药物输送、免疫和感染中的应用。由于粘蛋白是 在细胞防御的前线，不同的病毒已经进化出粘附其聚糖的策略，改变 它们，甚至用它们进入宿主细胞进行复制。病毒-粘蛋白结合会对病毒产生影响 扩散、组织特异性和复制，但由于粘蛋白异质性而缺乏分子细节。我们将 通过化学酶法修饰我们的 synMUC 以显示病毒结合唾液酸聚糖。病毒结合偏好 对于不同密度和不同肽主链组成的各种唾液酸结构，将是 定义的。带有唾液酸的 synMUC 将用于探测糖萼中的粘蛋白与粘液之间的关系 调节粘附、细胞进入和复制、组织向性和病毒基因进化。这些知识将会流失 阐明病毒生命周期的基本方面，可能有助于通过发展改善人类健康 新的抗病毒疗法和疾病传播预防策略。此外，科学公民 和指导是支持公平、多样性和包容性的优先事项和积极参与 STEM 中代表性不足的群体将成为项目持续期间及之后的重点。