Regulatory Interactions of CFTR and ENaC

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DESCRIPTION (provided by applicant): The Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a multifunctional protein that transports chloride across the apical plasma membrane of epithelial cells. CFTR also regulates ion transport by other proteins, such as the Epithelial Sodium Channel, ENaC. CFTR inhibits sodium transport by ENaC in airway epithelia, and one of the cardinal features of Cystic Fibrosis (CF) is hyperactivity of ENaC in the airway epithelia. However, the mechanism by which regulatory interactions of CFTR and ENaC occurs is not clear. Efforts at pharmacologic repair of mutant CFTR function have concentrated on assessing restoration of a mutant CFTR's chloride transport properties and have largely ignored other functions of CFTR, such as its interregulation with ENaC. It thus remains an open question whether pharmacologic repair of mutant CFTR function will also restore the critical regulatory interactions of CFTR and ENaC. Understanding the molecular basis of these critical regulatory interactions is therefore key in the implementation of pharmacologic strategies to improve CFTR function. Others have suggested that CFTR inhibits ENaC activity by decreasing ENaC open probability (Po). In contrast, our preliminary data strongly suggests that CFTR and ENaC may also have regulatory interactions related to intracellular trafficking. We will therefore test the hypothesis that: Regulatory interactions of CFTR and ENaC result in altered intracellular trafficking and surface expression of these channels in epithelial cells. Pharmacologic agents that repair mutant CFTR may modulate these trafficking interactions. Some such agents may influence trafficking by altering the expression of cytosolic molecular chaperones. As certain cytosolic domains of CFTR and ENaC appear critical for regulatory interactions, binding of chaperones to these cytosolic domains may regulate such interactions. The present proposal will build on these investigations and preliminary data and address this hypothesis with studies directed at the following Specific Aims: 1) To determine the kinetic mechanism by which CFTR and ENaC regulate each other's intracellular trafficking in epithelial cells. 2) To determine the mechanism by which cytosolic 70 kilodalton molecular chaperones influence and regulate these trafficking interactions of CFTR and ENaC in epithelial cells. Relevance: The major mechanism by which the lung and airway defends itself from the environment depends on proper ion transport in the respiratory epithelia. Such ion transport is aberrant in Cystic Fibrosis and leads to significant morbidity and mortality. These data will promote better understanding of the pathogenesis of not only Cystic Fibrosis lung disease, but also a number of other diseases of the airway, as well as diseases of other secretory epithelia.