Protein traffic disorders: an effective high-throughput fluorescence microscopy pipeline for drug discovery

Scientific Reports
Hugo M BotelhoMargarida D Amaral

Abstract

Plasma membrane proteins are essential molecules in the cell which mediate interactions with the exterior milieu, thus representing key drug targets for present pharma. Not surprisingly, protein traffic disorders include a large range of diseases sharing the common mechanism of failure in the respective protein to reach the plasma membrane. However, specific therapies for these diseases are remarkably lacking. Herein, we report a robust platform for drug discovery applied to a paradigmatic genetic disorder affecting intracellular trafficking - Cystic Fibrosis. This platform includes (i) two original respiratory epithelial cellular models incorporating an inducible double-tagged traffic reporter; (ii) a plasma membrane protein traffic assay for high-throughput microscopy screening; and (iii) open-source image analysis software to quantify plasma membrane protein traffic. By allowing direct scoring of compounds rescuing the basic traffic defect, this platform enables an effective drug development pipeline, which can be promptly adapted to any traffic disorder-associated protein and leverage therapy development efforts.

References

Apr 4, 1986·Science·M S Brown, J L Goldstein
Oct 16, 2001·American Journal of Physiology. Cell Physiology·L V GaliettaA S Verkman
Dec 25, 2002·Human Molecular Genetics·Cecile Lubrano-BerthelierChristian Vaisse
May 1, 1992·Trends in Cell Biology·J F AmaraA E Smith
May 6, 2004·Journal of Molecular Neuroscience : MN·Margarida D Amaral
Aug 30, 2005·The Journal of Clinical Investigation·Nicoletta PedemonteA S Verkman
Apr 22, 2006·Nature Methods·Beate NeumannJan Ellenberg
Mar 16, 2007·Proceedings of the National Academy of Sciences of the United States of America·Phillip FactorSadis Matalon
Apr 5, 2007·Nature Protocols·Holger ErfleRainer Pepperkok
Jul 5, 2008·Journal of Biomolecular Screening·Holger ErfleRainer Pepperkok
Apr 14, 2009·The Journal of Biological Chemistry·Jason R RockBarbara R Grubb
Aug 27, 2009·FEBS Letters·Darren M HuttWilliam E Balch
Feb 23, 2010·Proteomics·Linn FagerbergLisa Berglund
Jul 24, 2010·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·John HolleranJonathan W Jarvik
Nov 19, 2010·The New England Journal of Medicine·Frank J AccursoBonnie W Ramsey
Dec 3, 2010·The New England Journal of Medicine·John V Fahy, Burton F Dickey
Feb 9, 2011·Inflammation Research : Official Journal of the European Histamine Research Society ... [et Al.]·Xiao SuMichael A Matthay
Apr 13, 2011·Proceedings of the National Academy of Sciences of the United States of America·Claire L NewtonRobert P Millar
May 7, 2011·Methods in Molecular Biology·Joana AlmaçaMargarida D Amaral
May 20, 2011·Methods in Molecular Biology·Nicoletta PedemonteLuis J V Galietta
Sep 9, 2011·The New England Journal of Medicine·Maria Antonietta De Matteis, Alberto Luini
Oct 7, 2011·Proceedings of the National Academy of Sciences of the United States of America·Fredrick Van GoorPaul A Negulescu
Mar 20, 2012·Pediatric Nephrology : Journal of the International Pediatric Nephrology Association·Daniel WescheNine V A M Knoers
Jan 22, 2013·Current Pharmaceutical Design·Margarida D Amaral, Carlos M Farinha
Apr 11, 2013·Journal of Cell Science·John P HolleranSimon C Watkins
May 15, 2013·Annual Review of Genomics and Human Genetics·Maria Antonietta De MatteisCathal Wilson
Jan 1, 2014·Trends in Molecular Medicine·Bibek GooptuDavid A Lomas

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Citations

Nov 4, 2015·Journal of Cystic Fibrosis : Official Journal of the European Cystic Fibrosis Society·Margarida D Amaral, William E Balch
Nov 11, 2015·Human Mutation·Susana IgrejaMargarida D Amaral
May 6, 2016·Cellular and Molecular Life Sciences : CMLS·Joana F S PereiraPeter Jordan
Feb 7, 2018·Frontiers in Pharmacology·Sang Hyun LimIgor Stagljar
Jan 25, 2020·Cellular and Molecular Life Sciences : CMLS·Cláudia Almeida LoureiroPeter Jordan
Mar 28, 2019·Journal of Molecular Medicine : Official Organ of the Gesellschaft Deutscher Naturforscher Und Ärzte·Roberta BenedettoKarl Kunzelmann
Jun 25, 2019·Frontiers in Pharmacology·Cláudia Almeida LoureiroFrancisco R Pinto
Dec 14, 2018·Cellular and Molecular Life Sciences : CMLS·Waseema PatelMichael A Gray
Mar 1, 2017·Proceedings of the National Academy of Sciences of the United States of America·Zoltan BozokyJulie D Forman-Kay
May 26, 2018·Annual Review of Genomics and Human Genetics·Sarine MarkossianMichelle R Arkin
Aug 6, 2020·International Journal of Molecular Sciences·Nara LiessiClarissa Braccia
Sep 7, 2017·Pflügers Archiv : European journal of physiology·Filipa SimõesKarl Kunzelmann
Oct 1, 2017·Scientific Reports·Roberta BenedettoKarl Kunzelmann
Sep 17, 2020·The Journal of Biological Chemistry·Stella PrinsPaola Vergani
Nov 17, 2020·European Journal of Medicinal Chemistry·Margarida D Amaral
Jan 18, 2021·The Journal of Biological Chemistry·Stella PrinsPaola Vergani
Nov 5, 2019·Journal of Cystic Fibrosis : Official Journal of the European Cystic Fibrosis Society·Margarida D AmaralNicoletta Pedemonte
May 15, 2021·Frontiers in Pharmacology·Marjolein EnsinckMarianne S Carlon

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Methods Mentioned

BETA
protein folding
transfection

Software Mentioned

R
R script
CellProfiler

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