Mechanistic analysis of transport through the mucus barrier

产品说明：该项目的目标是获得深刻的机械理解和预测能力的分子机制，管理运输通过粘液屏障。粘液衬里形成一个选择性屏障，促进营养物质，精子和氧气的吸收，同时防止有害病毒，细菌和毒素的自由通过。区分穿过粘液屏障的颗粒和被粘液屏障排斥的颗粒的详细分子性质在很大程度上是未知的，因此，粘膜转运的预测模型目前缺失，尽管它们对药物递送和预防流行的传染病（例如乳头瘤病毒，HIV）具有巨大的影响。虽然已经单独研究了颗粒大小、净电荷和疏水性对粘液转运的相关性，但是还没有以允许预测粘液相互作用或设计具有定制粘液转运性质的药物递送载体的方式研究组合这些性质的效果和空间排列的作用。我们建议通过粘液屏障的分子运输的特点，并与电荷，疏水性和特定的肽序列的空间表面排列的结果。这种知识将使我们能够确定诊断快速和缓慢通过的生物物理指纹，并有可能改变药物递送载体的设计，因为它将允许联合收割机表面功能化（用于组织靶向），同时独立地调节载体的运输性质。在第一个目标中，我们将测试电荷分布和疏水性的影响，通过粘液运输使用短肽与系统不同的残基，和一个微流体系统来测量摄取，空间分布，并通过粘液运输。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（具体地，噬菌体）表面上的肽时，是否可以促进颗粒通过粘液。该系统还将深入了解影响颗粒-粘液相互作用的其他参数，例如肽长度、特定残基序列、表面显示密度和颗粒几何形状。在目标3中，我们将整合目标1和2的知识，并使用小鼠阴道作为模型，确定表面电荷、疏水性和特定肽序列影响体内粘液转运的相关性。多学科研究团队提供将基础科学问题与尖端工程应用相结合所需的专业知识：具有生物水凝胶系统的实验和理论专长的生物学家，具有生物组织中运输现象专长的机械工程师，具有工程噬菌体展示系统专长的合成生物学家，以及在受控颗粒表面功能化和体内表征方面具有专业知识的化学工程师。

DESCRIPTION: 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.