Regulation of epithelial function using targeted nanowires

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PROJECT SUMMARY The nasal cavity is a clinically accessible structure that is amenable to the topical application of nanoscale particles to facilitate drug delivery and control local inflammation. The success of this approach requires substrates with a high level of specificity and activity. We have designed and constructed derivatizable, biodegradable polycaprolactone (PCL) nanowires that are conjugated to anti-β1 integrin antibodies. Anti- integrin nanowires will be tested on cultured cell models, including well differentiated primary human lung and nasal airway epithelial cells, for the ability to alter barrier function, stimulate transcytosis, and affect cell growth and differentiation. Preliminary data supports our ability to develop anti-integrin nanowires with the capacity to either increase or decrease tight junction permeability, depending on composition. Several complementary approaches will be used to identify the mechanism of action for anti-integrin nanowires of different composition, focusing primarily on their effects on cytoplasmic scaffold proteins associated with tight junctions and integrins that regulate the actin cytoskeleton. This includes candidate and discovery based molecular methods to identify nanowire-driven changes to the cytoplasmic scaffold proteome. Live cell and super resolution microscopy will be used to measure the effects of anti-integrin nanowires on junctions and integrins in a native context. One goal is to establish modes of stimulating apical integrins as a pharmacologically tractable approach that can be preferentially switched to either improve respiratory epithelial barrier function or to facilitate drug delivery across epithelial barriers. We also will test the effects of anti-integrin nanowires on epithelia derived from nasal polyps, which have impaired tight junctions, are partially de-polarized, and lack growth control. These in vitro studies will evaluate nanowires as a potential platform to treat chronic rhinosinusitis with nasal polyposis, a chronic condition that significantly impairs quality of life and that is frequently associated with lung disease.