Project Number: 6066-21000-001-013-R
Project Type: Reimbursable Cooperative Agreement
Start Date: Oct 1, 2021
End Date: Jun 30, 2024
1. We will study the mechanisms of how cystic fibrosis transmembrane conductance regulator (CFTR) correctors (VX-661, VX-809, and analogs) interact with deltaF508-CFTR to improve the maturation and function of this mutant protein at the cell surface. 2. We will study the subject from the perspective of CFTR-containing macromolecular complex and employ click chemistry and photo-affinity labeling approaches.
The study will help us better understand the mechanisms of action of cystic fibrosis transmembrane conductance regulator (CFTR) correctors, expand our knowledge of CFTR-containing macromolecular complexes, and help identify novel therapeutic targets and develop more potent drugs for cystic fibrosis therapy. We propose to study (1) Does the temperature-rescued 'F508-CFTR form an unstable macromolecular complex at the plasma membrane? (2) Do CFTR correctors stabilize 'F508-CFTR at the plasma membrane by potentiating its interactions with binding partners and facilitating the formation of a stable macromolecular complex? (3) Do CFTR correctors bind directly to 'F508-CFTR? Does increasing their binding affinity produce a better rescue outcome? (4) Can we isolate the CFTR corrector-associated- and 'F508-CFTR-containing macromolecular complexes under different rescue conditions? By using proteomics, can we identify effectors and pathways important in the rescue process? We will generate derivatives of VX- CFTR correctors (named NN- CFTR correctors: NN-661, NN-809, NN-445, and NN-659) and use click chemistry and photo-affinity labeling to investigate the interactions and macromolecular complexes formation in various model systems.