Molecular Systems Biology (Feb 2022)
CFTR interactome mapping using the mammalian membrane two‐hybrid high‐throughput screening system
- Sang Hyun Lim,
- Jamie Snider,
- Liron Birimberg‐Schwartz,
- Wan Ip,
- Joana C Serralha,
- Hugo M Botelho,
- Miquéias Lopes‐Pacheco,
- Madalena C Pinto,
- Mohamed Taha Moutaoufik,
- Mara Zilocchi,
- Onofrio Laselva,
- Mohsen Esmaeili,
- Max Kotlyar,
- Anna Lyakisheva,
- Priscilla Tang,
- Lucía López Vázquez,
- Indira Akula,
- Farzaneh Aboualizadeh,
- Victoria Wong,
- Ingrid Grozavu,
- Teuta Opacak‐Bernardi,
- Zhong Yao,
- Meg Mendoza,
- Mohan Babu,
- Igor Jurisica,
- Tanja Gonska,
- Christine E Bear,
- Margarida D Amaral,
- Igor Stagljar
Affiliations
- Sang Hyun Lim
- Donnelly Centre University of Toronto Toronto ON Canada
- Jamie Snider
- Donnelly Centre University of Toronto Toronto ON Canada
- Liron Birimberg‐Schwartz
- Programme in Translational Medicine The Hospital for Sick Children Toronto ON Canada
- Wan Ip
- Programme in Translational Medicine The Hospital for Sick Children Toronto ON Canada
- Joana C Serralha
- Faculty of Sciences BioISI‐Biosystems and Integrative Sciences Institute University of Lisboa Lisboa Portugal
- Hugo M Botelho
- Faculty of Sciences BioISI‐Biosystems and Integrative Sciences Institute University of Lisboa Lisboa Portugal
- Miquéias Lopes‐Pacheco
- Faculty of Sciences BioISI‐Biosystems and Integrative Sciences Institute University of Lisboa Lisboa Portugal
- Madalena C Pinto
- Faculty of Sciences BioISI‐Biosystems and Integrative Sciences Institute University of Lisboa Lisboa Portugal
- Mohamed Taha Moutaoufik
- Department of Biochemistry, Research and Innovation Centre University of Regina Regina SK Canada
- Mara Zilocchi
- Department of Biochemistry, Research and Innovation Centre University of Regina Regina SK Canada
- Onofrio Laselva
- Department of Physiology University of Toronto Toronto ON Canada
- Mohsen Esmaeili
- Program in Genetics and Genome Biology The Hospital for Sick Children Toronto ON Canada
- Max Kotlyar
- Osteoarthritis Research Program Division of Orthopedic Surgery Schroeder Arthritis Institute University Health Network Toronto ON Canada
- Anna Lyakisheva
- Donnelly Centre University of Toronto Toronto ON Canada
- Priscilla Tang
- Donnelly Centre University of Toronto Toronto ON Canada
- Lucía López Vázquez
- Donnelly Centre University of Toronto Toronto ON Canada
- Indira Akula
- Donnelly Centre University of Toronto Toronto ON Canada
- Farzaneh Aboualizadeh
- Donnelly Centre University of Toronto Toronto ON Canada
- Victoria Wong
- Donnelly Centre University of Toronto Toronto ON Canada
- Ingrid Grozavu
- Donnelly Centre University of Toronto Toronto ON Canada
- Teuta Opacak‐Bernardi
- Faculty of Medicine Josip Juraj Strossmayer University of Osijek Osijek Croatia
- Zhong Yao
- Donnelly Centre University of Toronto Toronto ON Canada
- Meg Mendoza
- Department of Molecular Genetics University of Toronto Toronto ON Canada
- Mohan Babu
- Department of Biochemistry, Research and Innovation Centre University of Regina Regina SK Canada
- Igor Jurisica
- Osteoarthritis Research Program Division of Orthopedic Surgery Schroeder Arthritis Institute University Health Network Toronto ON Canada
- Tanja Gonska
- Programme in Translational Medicine The Hospital for Sick Children Toronto ON Canada
- Christine E Bear
- Department of Biochemistry University of Toronto Toronto ON Canada
- Margarida D Amaral
- Faculty of Sciences BioISI‐Biosystems and Integrative Sciences Institute University of Lisboa Lisboa Portugal
- Igor Stagljar
- Donnelly Centre University of Toronto Toronto ON Canada
- DOI
- https://doi.org/10.15252/msb.202110629
- Journal volume & issue
-
Vol. 18,
no. 2
pp. n/a – n/a
Abstract
Abstract Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a chloride and bicarbonate channel in secretory epithelia with a critical role in maintaining fluid homeostasis. Mutations in CFTR are associated with Cystic Fibrosis (CF), the most common lethal autosomal recessive disorder in Caucasians. While remarkable treatment advances have been made recently in the form of modulator drugs directly rescuing CFTR dysfunction, there is still considerable scope for improvement of therapeutic effectiveness. Here, we report the application of a high‐throughput screening variant of the Mammalian Membrane Two‐Hybrid (MaMTH‐HTS) to map the protein–protein interactions of wild‐type (wt) and mutant CFTR (F508del), in an effort to better understand CF cellular effects and identify new drug targets for patient‐specific treatments. Combined with functional validation in multiple disease models, we have uncovered candidate proteins with potential roles in CFTR function/CF pathophysiology, including Fibrinogen Like 2 (FGL2), which we demonstrate in patient‐derived intestinal organoids has a significant effect on CFTR functional expression.
Keywords
- cystic fibrosis
- high‐throughput screening
- integrative computational biology
- interactome
- mammalian membrane two‐hybrid