Synthetic mucins in epithelial models to probe virus-mucin interactions

项目总结 粘蛋白糖蛋白是粘液和上皮细胞糖萼的基本成分。粘蛋白是 对从水母到人类的生物都是必不可少的，并在水合、润滑、营养吸收、 以及宿主对病原体的防御。粘蛋白糖基化受复杂的酶途径调节 以流动，导致不同组织和物种之间不同的和可变的糖链模式，并且 随着饮食和疾病的变化而进化。克莱默实验室正在开发合成粘蛋白，或称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-羧酸酐聚合可制得高分子量多肽 天然多肽和葡聚糖的连接。与短肽、多糖或传统聚合物相比 SynMUC含有附着糖，是迄今为止最真实的粘蛋白模拟物。将应用synMUC 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 上皮生物学的研究及其在癌症、药物输送、免疫和感染中的应用。因为粘蛋白是 在细胞防御的前线，各种病毒已经进化出附着在它们的糖链上的策略，改变了 甚至利用它们进入宿主细胞进行复制。病毒-粘蛋白结合可以在病毒上产生结果 扩散、组织特异性和复制，但由于粘蛋白的异质性，缺乏分子细节。我们会 化学酶修饰我们的synMUC以显示与病毒结合的唾液酸多糖。病毒结合偏好 对于不同密度的不同唾液酸结构和来自不同的肽骨架组合物 已定义。含唾液酸的synMUC将被用来探测糖萼中的粘蛋白与粘液是如何 调节黏附、细胞进入和复制、组织趋向性和病毒基因进化。这一知识将会抛弃 阐明病毒生命周期的基本方面，并可能有助于通过发展来改善人类健康 新的抗病毒疗法和疾病传播预防战略。此外，科学公民身份 和辅导是支持公平、多样性和包容性的优先事项和积极参与 在STEM中任职人数不足的群体将是项目期间及以后的重点。

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.