Mechanistic analysis of transport through the mucus barrier

项目摘要 这个项目的目标是获得对分子的深入的机械理解和预测能力。 控制粘液屏障运输的机制。粘液衬里形成选择性屏障， 促进营养物质、精子和氧气的吸收，同时防止有害病毒的自由传播， 细菌和毒素。区分通过粘液的颗粒的详细分子特性 被粘液屏障排斥的屏障和颗粒在很大程度上是未知的，因此，预测模型 目前尚不清楚，尽管它们对药物输送和 预防流行的传染病(如乳头瘤病毒、艾滋病毒)。而粒子的关联性 粘液传输的大小、净电荷和疏水性已经在隔离中进行了研究，结合 这些属性和空间排列的作用还没有以允许预测的方式进行研究 粘液相互作用或设计具有粘液传输特性的药物输送载体。我们建议 表征分子通过粘液屏障的传输，并将结果与空间表面联系起来 电荷排列、疏水性和特定的多肽序列。这一知识将使我们能够 确定对快速和缓慢通过进行诊断的生物物理指纹，并有可能 改变药物输送载体的设计，因为它将允许结合表面功能化(用于组织 目标)，同时独立地调整车辆的运输属性。在第一个目标中，我们将测试 短肽对粘液转运过程中电荷分布和疏水性的影响 系统变化的残留物，以及测量摄取、空间分布和传输的微流控系统 通过粘液。在第二个目标中，我们将使用基于噬菌体展示的方法来确定 这些相同的规则，当应用于颗粒表面的多肽(具体地说，噬菌体)时，可以促进 颗粒通过粘液的过程。该系统还将提供对影响 颗粒-粘液相互作用，如多肽长度、特定残基序列、表面显示密度和 粒子几何学。在目标3中，我们将整合来自目标1和目标2的知识，并确定 表面电荷、疏水性和特定的多肽序列影响体内粘液的运输，使用 以小鼠阴道为模型。多学科研究团队提供了将 具有尖端工程应用的基础科学问题：一位生物学家 生物水凝胶系统的理论专业知识，具有运输专业知识的机械工程师 生物组织中的现象，具有工程噬菌体展示系统专业知识的合成生物学家，以及 一位在受控粒子表面功能化和体内表征方面具有专业知识的化学工程师。

Project Summary The goal of this project is to gain deep mechanistic understanding and predictive capability of the molecular mechanisms that govern transport through the mucus barrier. The mucus lining forms a selective barrier that facilitates the uptake of nutrients, sperm, and oxygen, while preventing free passage of harmful viruses, bacteria, and toxins. The detailed molecular properties that distinguish particles that pass through the mucus barrier and particles that are rejected by the mucus barrier are largely unknown, and hence, predictive models for mucosal transport are currently missing, despite their tremendous implications for drug delivery and preventing prevalent infectious diseases (for example Papilloma virus, HIV). While the relevance of particle size, net charge, and hydrophobicity for mucus transport has been studied in isolation, the effect of combining these properties and the role of spatial arrangement have not been studied in a way that allows to predict mucus-interactions or to design drug delivery vehicles with tailored mucus transport properties. We propose to characterize molecular transport through the mucus barrier and relate the results to the spatial surface arrangement of charge, hydrophobicity and specific peptide sequences. This knowledge will enable us to determine biophysical fingerprints that are diagnostic for fast and slow passage, and has the potential to transform the design of drug delivery vehicles as it will allow to combine surface functionalization (for tissue targeting) while independently tuning the transport properties of a vehicle. In the first aim we will test the influence of charge distribution and hydrophobicity for transport through mucus using short peptides with systematically varied residues, and a microfluidic system to measure uptake, spatial distribution, and transport through the mucus. In the second aim, we will use phage-display-based approaches to determine whether these same rules, when applied to peptides on the surface of a particle (specifically, phage) can facilitate passage of the particle through mucus. This system will also give insight into other parameters that affect particle-mucus interactions, such as peptide length, specific residue sequence, surface display density, and particle geometry. In Aim 3 we will integrate the knowledge from Aims 1 and 2 and determine the relevance of surface charge, hydrophobicity, and specific peptide sequences influences mucus transport in vivo, using the mouse vagina as a model. The multidisciplinary research team presents the expertise necessary for combining fundamental science questions with cutting edge engineering applications: a biologist with experimental and theoretical expertise in biological hydrogel systems, a mechanical engineer with expertise in transport phenomena in biological tissues, a synthetic biologist with expertise in engineering phage display systems, and a chemical engineer with expertise in controlled particle surface functionalization and characterization in vivo.