%0 Journal Article %A Andrew G. McKee %A Eli F. McDonald %A Wesley D. Penn %A Charles P. Kuntz %A Karen Noguera %A Laura M. Chamness %A Francis J. Roushar %A Jens Meiler %A Kathryn E. Oliver %A Lars Plate %A Jonathan P. Schlebach %T Elucidation of Global Trends in the Effects of VX-661 and VX-445 on the Expression of Clinical CFTR Variants %D 2022 %R 10.1101/2022.10.14.512300 %J bioRxiv %P 2022.10.14.512300 %X Cystic fibrosis (CF) is a chronic genetic disease caused by mutations that compromise the expression and/ or function of the cystic fibrosis transmembrane conductance regulator chloride channel (CFTR). Most people with CF harbor a common misfolded CFTR variant (ΔF508), which can be rescued by combination therapies containing “corrector” compounds that restore its expression. Nevertheless, there are over 400 other CF variants that differ in their sensitivity to correctors for reasons that remain unclear. In this work, we utilize deep mutational scanning to quantitatively compare the effects of two FDA-approved correctors on the plasma membrane expression of 129 known CF variants, including 45 that are currently unclassified. Across 67 variants with attenuated expression, we find that VX-661-sensitive variants generally exhibit intermediate expression and feature mutations near its binding pocket within the first membrane spanning domain (MSD1). VX-445 also rescues variants with intermediate expression, though it is uniquely effective for certain mutations that perturb the later stages of CFTR assembly. Structural calculations suggest VX-661 provides similar stabilization to both sensitive and insensitive variants alike. These findings collectively suggest the mutation-specific effects of these compounds depend on the degree of variant destabilization and/ or the timing of cotranslational folding defects. Combining these correctors synergistically rescues variants with deficient and intermediate expression alike, presumably by doubling the total binding energy and suppressing defects at different stages of translation. These results provide an unprecedented overview of the properties of rare CFTR variants and establish new tools for CF pharmacology.Competing Interest StatementThe authors have declared no competing interest. %U https://www.biorxiv.org/content/biorxiv/early/2022/10/17/2022.10.14.512300.full.pdf