Proximity Profiling of the CFTR Interaction Landscape in Response to Orkambi

Deletion of phenylalanine 508 ( increment F508) of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) anion channel protein is the leading cause of Cystic Fibrosis (CF). Here, we report the analysis of CFTR and increment F508-CFTR interactomes using BioID (proximity-dependent biotin identification), a technique that can also detect transient associations. We identified 474 high-confidence CFTR proximity-interactors, 57 of which have been previously validated, with the remainder representing novel interaction space. The increment F508 interactome, comprising 626 proximity-interactors was markedly different from its wild type counterpart, with numerous alterations in protein associations categorized in membrane trafficking and cellular stress functions. Furthermore, analysis of the increment F508 interactome in cells treated with Orkambi identified several interactions that were altered as a result of this drug therapy. We examined two candidate CFTR proximity interactors, VAPB and NOS1AP, in functional assays designed to assess surface delivery and overall chloride efflux. VAPB depletion impacted both CFTR surface delivery and chloride efflux, whereas NOS1AP depletion only affected the latter. The wild type and increment F508-CFTR interactomes represent rich datasets that could be further mined to reveal additional candidates for the functional rescue of increment F508-CFTR.

Automated summary

Deletion of phenylalanine 508 (increment F508) of the CFTR protein is the main cause of CF. This study analyzed the CFTR and increment F508-CFTR interactomes using BioID, and identified known and novel protein interactions. The increment F508 interactome showed significant differences compared to the wild type, with alterations in membrane trafficking and cellular stress functions. Drug therapy also affected the interactome, and depletion of specific interactors impacted CFTR functionality.