CFTR interactome mapping using the mammalian membrane two-hybrid high-throughput screening system.

Area of Science:

• Biochemistry and Molecular Biology
• Genetics and Genomics
• Physiology

Background:

• Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a vital ion channel in epithelial cells.
• Mutations in CFTR cause Cystic Fibrosis (CF), a severe genetic disorder.
• Current CFTR modulator therapies show promise but require further enhancement.

Purpose of the Study:

• To map protein-protein interactions of wild-type and F508del mutant CFTR.
• To identify novel drug targets for patient-specific CF treatments.
• To understand CF cellular effects and improve therapeutic strategies.

Main Methods:

• Utilized a high-throughput screening variant of the Mammalian Membrane Two-Hybrid (MaMTH-HTS) system.
• Mapped protein interactions for both wild-type (wt) and F508del mutant CFTR.
• Performed functional validation in multiple disease models, including patient-derived intestinal organoids.

Main Results:

• Successfully mapped CFTR protein-protein interactions.
• Identified Fibrinogen Like 2 (FGL2) as a candidate protein involved in CFTR function.
• Demonstrated that FGL2 significantly affects CFTR functional expression in patient-derived organoids.

Conclusions:

• The MaMTH-HTS approach is effective for identifying CFTR interacting proteins.
• FGL2 emerges as a promising therapeutic target for enhancing CFTR function.
• Findings support the development of novel, patient-specific treatments for Cystic Fibrosis.