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Intratracheal Aerosolization of Viral Vectors to Newborn Pig Airways

Ashley L. Cooney1, Patrick L. Sinn1

Abstract

Gene therapy for airway diseases requires efficient delivery of nucleic acids to the intrapulmonary airways. In small animal models such as mice, gene delivery reagents are commonly delivered as a bolus dose. Routes of delivery may include either nasal sniffing or direct tracheal instillation. However, using a large animal model for preclinical applications is relevant for translation to human trials. Widespread and uniform distribution of transgene expression is critical for developing a successful lung gene therapy treatment. Aerosolizing viral vectors to the lungs of large animals, such as pigs or sheep, is a strategy to maximize gene transfer efficiency and results in greater airway distribution than a liquid bolus dose. Here we demonstrate a technique for direct aerosolization of a viral vector to the airways of newborn pigs. Briefly, a pig is anesthetized, intubated with an endotracheal tube, and a microsprayer is passed through the endotracheal tube. A syringe is used to push the vector through the microsprayer, resulting in a fine mist being released into the distal trachea. The microsprayer produces ~15-16 μm size particles that deposit across the proximal and distal regions of the lung. Using a microsprayer to deliver an adenoviral-based vector, we previously observed ~30-50% of surface epithelial cells transduced in both the large and small airways of newborn pigs.

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