PROJECT ABSTRACT Voice disorders are among the most common communication disorders and are often associated with mucosa inflammatory etiologies. Furthermore, laryngeal sensory disorders represent a significant clinical cohort that present ubiquitous treatment challenges. Recent discovery of mechanosensitive ion channels, Piezo1 and Piezo2, has proved essential to development and homeostasis for various tissue phenotypes that undergo mechanical loading (e.g. lung, skin, cartilage, bladder, vasculature, heart). As molecular transducers of mechanical force, Piezo proteins are activated by mechanical stretch stimuli exerted on cell membranes, upon which they rapidly and efficiently convert stimuli into intracellular signaling events for biologic effect. Evidence supports Piezo1 presence and function in epithelial cells, however, the role of Piezo1 in vocal fold (VF) – a tissue type that undergoes extreme biomechanical challenges, is not known. In this application, a series of experiments to determine the contribution of Piezo1 to vocal fold epithelium development and function is proposed. We will employ two different in vivo strategies to delete/knockout (KO) Piezo1 using cell specific carriers. Aim 1 will define Piezo1 regulation in vocal fold epithelium morphogenesis and structural barrier protection mechanisms following widespread Piezo1 KO in VF mucosa. Aim 2 will determine Piezo1 function localized to VF basal epithelial cells via Keratin 14 promoter to elucidate differentiation and self-renewal strategies. Our overarching hypothesis is that Piezo1 is a critical mechanotransducer for VF epithelium; essential for a matured, stratified tissue architecture with necessary barrier function. This innovative application will provide vital knowledge for mechanism(s) that govern cell signaling pathways involved in mechanosensory transduction processes in the developing and adult VF epithelium.