Mechanistic analysis of transport through the mucus barrier

描述：这个项目的目标是获得通过黏液屏障控制运输的分子机制的深刻机制理解和预测能力。粘液层形成选择性屏障，促进营养物质、精子和氧气的吸收，同时防止有害病毒、细菌和毒素的自由通过。区分通过粘液屏障的颗粒和被粘液屏障拒绝的颗粒的详细分子特性在很大程度上是未知的，因此，粘膜运输的预测模型目前是缺失的，尽管它们对药物输送和预防流行的传染病（例如乳头状瘤病毒，HIV）具有巨大的意义。虽然已经单独研究了颗粒大小、净电荷和疏水性与黏液运输的相关性，但结合这些特性的影响和空间排列的作用还没有得到研究，从而无法预测黏液相互作用或设计具有定制黏液运输特性的药物输送载体。我们建议表征分子通过黏液屏障的运输，并将结果与电荷、疏水性和特定肽序列的空间表面排列联系起来。这些知识将使我们能够确定生物物理指纹，用于诊断快速和慢速通道，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（用于组织靶向），同时独立调整载体的运输特性。在第一个目标中，我们将测试电荷分布和疏水性对粘液运输的影响，使用具有系统变化残基的短肽，以及微流体系统来测量吸收，空间分布和通过粘液的运输。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（特别是噬菌体）表面的肽时，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-黏液相互作用的其他参数，如肽长度、特定残基序列、表面显示密度和颗粒几何形状。在Aims 3中，我们将整合Aims 1和Aims 2的知识，并以小鼠阴道为模型，确定表面电荷、疏水性和特定肽序列在体内影响粘液运输的相关性。多学科研究团队展示了将基础科学问题与前沿工程应用相结合所必需的专业知识：在生物水凝胶系统方面具有实验和理论专长的生物学家，在生物组织中运输现象方面具有专长的机械工程师，在工程噬菌体展示系统方面具有专长的合成生物学家，以及在体内控制颗粒表面功能化和表征方面具有专长的化学工程师。

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.