title: Latonduine Analogs Restore F508del-Cystic Fibrosis Transmembrane Conductance Regulator Trafficking through the Modulation of Poly-ADP Ribose Polymerase 3 and Poly-ADP Ribose Polymerase 16 Activity.
abstract: Cystic fibrosis (CF) is a major lethal genetic disease caused by mutations in the CF transmembrane conductance regulator gene (CFTR). This encodes a chloride ion channel on the apical surface of epithelial cells. The most common mutation in CFTR (F508del-CFTR) generates a protein that is misfolded and retained in the endoplasmic reticulum. Identifying small molecules that correct this CFTR trafficking defect is a promising approach in CF therapy. However, to date only modest efficacy has been reported for correctors in clinical trials. We identified the marine sponge metabolite latonduine as a corrector. We have now developed a series of latonduine derivatives that are more potent F508del-CFTR correctors with one (MCG315 [2,3-dihydro-1H-2-benzazepin-1-one]) having 10-fold increased corrector activity and an EC50 of 72.25 nM. We show that the latonduine analogs inhibit poly-ADP ribose polymerase (PARP) isozymes 1, 3, and 16. Further our molecular modeling studies point to the latonduine analogs binding to the PARP nicotinamide-binding domain. We established the relationship between the ability of the latonduine analogs to inhibit PARP-16 and their ability to correct F508del-CFTR trafficking. We show that latonduine can inhibit both PARP-3 and -16 and that this is necessary for CFTR correction. We demonstrate that latonduine triggers correction by regulating the activity of the unfolded protein response activator inositol-requiring enzyme (IRE-1) via modulation of the level of its ribosylation by PARP-16. These results establish latonduines novel site of action as well as its proteostatic mechanism of action.
authors.affiliation: Department of Biochemistry (G.W.C., R.R., Q.Y., D.Y.T.), Cystic Fibrosis Translational Research Center (G.W.C., R.R., E.M., Q.Y., J.W.H., D.Y.T.), and Department of Physiology (E.M., J.W.H.), McGill University, Montréal, Québec, Canada; GSK Respiratory Therapeutic Area Unit (R.S., R.H., V.B.), and R&D Platform Technology and Science (C.M.E.), GlaxoSmithKline Medicines Research Centre, Stevenage, Hertfordshire, United Kingdom; Departments of Chemistry and Earth, Ocean and Atmospheric Sciences, University of British Columbia, Vancouver, British Columbia, Canada (R.A.); Faculty of Medicine National Heart and Lung Institute, Imperial College London, London, United Kingdom (R.S.); Francis Crick Institute, London, United Kingdom (V.B.) graeme.carlile@McGill.ca.
authors.forename: Graeme W
authors.initials: GW
authors.lastname: Carlile
authors.affiliation: Department of Biochemistry (G.W.C., R.R., Q.Y., D.Y.T.), Cystic Fibrosis Translational Research Center (G.W.C., R.R., E.M., Q.Y., J.W.H., D.Y.T.), and Department of Physiology (E.M., J.W.H.), McGill University, Montréal, Québec, Canada; GSK Respiratory Therapeutic Area Unit (R.S., R.H., V.B.), and R&D Platform Technology and Science (C.M.E.), GlaxoSmithKline Medicines Research Centre, Stevenage, Hertfordshire, United Kingdom; Departments of Chemistry and Earth, Ocean and Atmospheric Sciences, University of British Columbia, Vancouver, British Columbia, Canada (R.A.); Faculty of Medicine National Heart and Lung Institute, Imperial College London, London, United Kingdom (R.S.); Francis Crick Institute, London, United Kingdom (V.B.).
authors.forename: Renaud
authors.initials: R
authors.lastname: Robert
authors.affiliation: Department of Biochemistry (G.W.C., R.R., Q.Y., D.Y.T.), Cystic Fibrosis Translational Research Center (G.W.C., R.R., E.M., Q.Y., J.W.H., D.Y.T.), and Department of Physiology (E.M., J.W.H.), McGill University, Montréal, Québec, Canada; GSK Respiratory Therapeutic Area Unit (R.S., R.H., V.B.), and R&D Platform Technology and Science (C.M.E.), GlaxoSmithKline Medicines Research Centre, Stevenage, Hertfordshire, United Kingdom; Departments of Chemistry and Earth, Ocean and Atmospheric Sciences, University of British Columbia, Vancouver, British Columbia, Canada (R.A.); Faculty of Medicine National Heart and Lung Institute, Imperial College London, London, United Kingdom (R.S.); Francis Crick Institute, London, United Kingdom (V.B.).
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Matthes
authors.affiliation: Department of Biochemistry (G.W.C., R.R., Q.Y., D.Y.T.), Cystic Fibrosis Translational Research Center (G.W.C., R.R., E.M., Q.Y., J.W.H., D.Y.T.), and Department of Physiology (E.M., J.W.H.), McGill University, Montréal, Québec, Canada; GSK Respiratory Therapeutic Area Unit (R.S., R.H., V.B.), and R&D Platform Technology and Science (C.M.E.), GlaxoSmithKline Medicines Research Centre, Stevenage, Hertfordshire, United Kingdom; Departments of Chemistry and Earth, Ocean and Atmospheric Sciences, University of British Columbia, Vancouver, British Columbia, Canada (R.A.); Faculty of Medicine National Heart and Lung Institute, Imperial College London, London, United Kingdom (R.S.); Francis Crick Institute, London, United Kingdom (V.B.).
authors.forename: Qi
authors.initials: Q
authors.lastname: Yang
authors.affiliation: Department of Biochemistry (G.W.C., R.R., Q.Y., D.Y.T.), Cystic Fibrosis Translational Research Center (G.W.C., R.R., E.M., Q.Y., J.W.H., D.Y.T.), and Department of Physiology (E.M., J.W.H.), McGill University, Montréal, Québec, Canada; GSK Respiratory Therapeutic Area Unit (R.S., R.H., V.B.), and R&D Platform Technology and Science (C.M.E.), GlaxoSmithKline Medicines Research Centre, Stevenage, Hertfordshire, United Kingdom; Departments of Chemistry and Earth, Ocean and Atmospheric Sciences, University of British Columbia, Vancouver, British Columbia, Canada (R.A.); Faculty of Medicine National Heart and Lung Institute, Imperial College London, London, United Kingdom (R.S.); Francis Crick Institute, London, United Kingdom (V.B.).
authors.forename: Roberto
authors.initials: R
authors.lastname: Solari
authors.affiliation: Department of Biochemistry (G.W.C., R.R., Q.Y., D.Y.T.), Cystic Fibrosis Translational Research Center (G.W.C., R.R., E.M., Q.Y., J.W.H., D.Y.T.), and Department of Physiology (E.M., J.W.H.), McGill University, Montréal, Québec, Canada; GSK Respiratory Therapeutic Area Unit (R.S., R.H., V.B.), and R&D Platform Technology and Science (C.M.E.), GlaxoSmithKline Medicines Research Centre, Stevenage, Hertfordshire, United Kingdom; Departments of Chemistry and Earth, Ocean and Atmospheric Sciences, University of British Columbia, Vancouver, British Columbia, Canada (R.A.); Faculty of Medicine National Heart and Lung Institute, Imperial College London, London, United Kingdom (R.S.); Francis Crick Institute, London, United Kingdom (V.B.).
authors.forename: Richard
authors.initials: R
authors.lastname: Hatley
authors.affiliation: Department of Biochemistry (G.W.C., R.R., Q.Y., D.Y.T.), Cystic Fibrosis Translational Research Center (G.W.C., R.R., E.M., Q.Y., J.W.H., D.Y.T.), and Department of Physiology (E.M., J.W.H.), McGill University, Montréal, Québec, Canada; GSK Respiratory Therapeutic Area Unit (R.S., R.H., V.B.), and R&D Platform Technology and Science (C.M.E.), GlaxoSmithKline Medicines Research Centre, Stevenage, Hertfordshire, United Kingdom; Departments of Chemistry and Earth, Ocean and Atmospheric Sciences, University of British Columbia, Vancouver, British Columbia, Canada (R.A.); Faculty of Medicine National Heart and Lung Institute, Imperial College London, London, United Kingdom (R.S.); Francis Crick Institute, London, United Kingdom (V.B.).
authors.forename: Colin M
authors.initials: CM
authors.lastname: Edge
authors.affiliation: Department of Biochemistry (G.W.C., R.R., Q.Y., D.Y.T.), Cystic Fibrosis Translational Research Center (G.W.C., R.R., E.M., Q.Y., J.W.H., D.Y.T.), and Department of Physiology (E.M., J.W.H.), McGill University, Montréal, Québec, Canada; GSK Respiratory Therapeutic Area Unit (R.S., R.H., V.B.), and R&D Platform Technology and Science (C.M.E.), GlaxoSmithKline Medicines Research Centre, Stevenage, Hertfordshire, United Kingdom; Departments of Chemistry and Earth, Ocean and Atmospheric Sciences, University of British Columbia, Vancouver, British Columbia, Canada (R.A.); Faculty of Medicine National Heart and Lung Institute, Imperial College London, London, United Kingdom (R.S.); Francis Crick Institute, London, United Kingdom (V.B.).
authors.forename: John W
authors.initials: JW
authors.lastname: Hanrahan
authors.affiliation: Department of Biochemistry (G.W.C., R.R., Q.Y., D.Y.T.), Cystic Fibrosis Translational Research Center (G.W.C., R.R., E.M., Q.Y., J.W.H., D.Y.T.), and Department of Physiology (E.M., J.W.H.), McGill University, Montréal, Québec, Canada; GSK Respiratory Therapeutic Area Unit (R.S., R.H., V.B.), and R&D Platform Technology and Science (C.M.E.), GlaxoSmithKline Medicines Research Centre, Stevenage, Hertfordshire, United Kingdom; Departments of Chemistry and Earth, Ocean and Atmospheric Sciences, University of British Columbia, Vancouver, British Columbia, Canada (R.A.); Faculty of Medicine National Heart and Lung Institute, Imperial College London, London, United Kingdom (R.S.); Francis Crick Institute, London, United Kingdom (V.B.).
authors.forename: Raymond
authors.initials: R
authors.lastname: Andersen
authors.affiliation: Department of Biochemistry (G.W.C., R.R., Q.Y., D.Y.T.), Cystic Fibrosis Translational Research Center (G.W.C., R.R., E.M., Q.Y., J.W.H., D.Y.T.), and Department of Physiology (E.M., J.W.H.), McGill University, Montréal, Québec, Canada; GSK Respiratory Therapeutic Area Unit (R.S., R.H., V.B.), and R&D Platform Technology and Science (C.M.E.), GlaxoSmithKline Medicines Research Centre, Stevenage, Hertfordshire, United Kingdom; Departments of Chemistry and Earth, Ocean and Atmospheric Sciences, University of British Columbia, Vancouver, British Columbia, Canada (R.A.); Faculty of Medicine National Heart and Lung Institute, Imperial College London, London, United Kingdom (R.S.); Francis Crick Institute, London, United Kingdom (V.B.).
authors.forename: David Y
authors.initials: DY
authors.lastname: Thomas
authors.affiliation: Department of Biochemistry (G.W.C., R.R., Q.Y., D.Y.T.), Cystic Fibrosis Translational Research Center (G.W.C., R.R., E.M., Q.Y., J.W.H., D.Y.T.), and Department of Physiology (E.M., J.W.H.), McGill University, Montréal, Québec, Canada; GSK Respiratory Therapeutic Area Unit (R.S., R.H., V.B.), and R&D Platform Technology and Science (C.M.E.), GlaxoSmithKline Medicines Research Centre, Stevenage, Hertfordshire, United Kingdom; Departments of Chemistry and Earth, Ocean and Atmospheric Sciences, University of British Columbia, Vancouver, British Columbia, Canada (R.A.); Faculty of Medicine National Heart and Lung Institute, Imperial College London, London, United Kingdom (R.S.); Francis Crick Institute, London, United Kingdom (V.B.).
authors.forename: Véronique
authors.initials: V
authors.lastname: Birault
mesh_terms: D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D000818:Animals; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000378:metabolism*; D002460:Cell Line; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000378:metabolism*; D004722:Endoribonucleases / Q000378:metabolism; D055785:Gene Knockdown Techniques; D006023:Glycoproteins / Q000378:metabolism; D006575:Heterocyclic Compounds, 3-Ring / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D008958:Models, Molecular; D000067856:Poly(ADP-ribose) Polymerase Inhibitors / Q000737:chemistry / Q000494:pharmacology; D011065:Poly(ADP-ribose) Polymerases / Q000737:chemistry / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects; D034741:RNA, Small Interfering / Q000378:metabolism; D056811:Unfolded Protein Response / Q000187:drug effects
chemical_list: D018797:Cell Cycle Proteins; D006023:Glycoproteins; D006575:Heterocyclic Compounds, 3-Ring; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D034741:RNA, Small Interfering; C093349:cystic fibrosis transmembrane conductance regulator delta F508; C478428:latonduine A; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D000246:Adenosine Diphosphate Ribose; C576117:PARP16 protein, human; C480979:PARP3 protein, human; D011065:Poly(ADP-ribose) Polymerases; D004722:Endoribonucleases