Pharmacological inhibition of the cystic fibrosis transmembrane conductance regulator (CFTR)

Weill Medical Coll Of Cornell Univ, New York NY

Investigators

Abstract

PROJECT SUMMARY/ABSTRACT Autosomal dominant polycystic kidney disease (ADPKD) is the most common inherited kidney disease and fourth leading cause of end-stage renal disease (ESRD). Hyperactivation of the cystic fibrosis transmembrane conductance regulator (CFTR) is essential for cyst formation and enlargement in ADPKD, and inhibition of CFTR is a potential therapeutic strategy. Despite its importance as a pharmacological target, the conformations transited by CFTR during physiological gating are poorly resolved, and the mechanism by which nucleotide state is communicated from the nucleotide binding domains (NBDs) to the pore 50 Å away remains obscure. Additionally, molecular mechanisms of pharmacological inhibition have been understudied despite therapeutic promise for ADPKD. Specific aim 1 seeks to determine the structural basis of allosteric communication between the NBDs and the pore using cryogenic electron microscopy (cryo-EM). Specific aim 2 seeks to determine the mechanisms of existing pharmacological inhibitors of CFTR using cryo-EM in combination with electrophysiological, biochemical, and biophysical assays. Specific aim 3 seeks to find new inhibitors of CFTR to enhance treatment of ADPKD using computational docking and medicinal chemistry coupled with high-throughput assays, electrophysiology, and mouse models of ADPKD. Successful completion of these goals will fill a gap in our understanding of the CFTR gating cycle, provide new conformations for pharmacological targeting, expand our understanding of CFTR’s pharmacological regulation, and establish new leads for treatment of ADPKD. The proposed research strategy will be completed under the mentorship of Dr. Jue Chen in the Laboratory of Membrane Biology and Biophysics at The Rockefeller University (RU) in New York City with the co-advisement of Dr. Jiankun Lyu at RU. The accompanying fellowship training plan will be completed through the Tri- Institutional MD-PhD Program of Weill Cornell Medicine, The Rockefeller University, and Memorial Sloan Kettering Cancer Center. The exceptional clinical and scientific resources of these environments will facilitate successful completion of both research and training components. The electrophysiological, biochemical, biophysical, and structural biology equipment and expertise available at The Rockefeller University is unparalleled, and trainees are encouraged to collaborate with other scientists and institutions in New York City’s rich structural biology research network. The Tri-Institutional MD-PhD Program offers a diversity of clinical, research, and extracurricular opportunities for trainees to develop the analytical, technical, communication, and mentorship skills necessary to become independent physician-scientists. Trainees are encouraged to continuously develop their clinical and scientific skills through both medical and graduate phases of the MD-PhD curriculum. This curriculum is supervised by a supportive administration that provides ample guidance for trainees to pursue productive careers promoting the health of the American public.