title: Matrix metalloproteinase 2 (MMP-2) degrades soluble vasculotropic amyloid-beta E22Q and L34V mutants, delaying their toxicity for human brain microvascular endothelial cells.
abstract: Patients carrying mutations within the amyloid-beta (Abeta) sequence develop severe early-onset cerebral amyloid angiopathy with some of the related variants manifesting primarily with hemorrhagic phenotypes. Matrix metalloproteases (MMPs) are typically associated with blood brain barrier disruption and hemorrhagic transformations after ischemic stroke. However, their contribution to cerebral amyloid angiopathy-related hemorrhage remains unclear. Human brain endothelial cells challenged with Abeta synthetic homologues containing mutations known to be associated in vivo with hemorrhagic manifestations (AbetaE22Q and AbetaL34V) showed enhanced production and activation of MMP-2, evaluated via Multiplex MMP antibody arrays, gel zymography, and Western blot, which in turn proteolytically cleaved in situ the Abeta peptides. Immunoprecipitation followed by mass spectrometry analysis highlighted the generation of specific C-terminal proteolytic fragments, in particular the accumulation of Abeta-(1-16), a result validated in vitro with recombinant MMP-2 and quantitatively evaluated using deuterium-labeled internal standards. Silencing MMP-2 gene expression resulted in reduced Abeta degradation and enhanced apoptosis. Secretion and activation of MMP-2 as well as susceptibility of the Abeta peptides to MMP-2 degradation were dependent on the peptide conformation, with fibrillar elements of AbetaE22Q exhibiting negligible effects. Our results indicate that MMP-2 release and activation differentially degrades Abeta species, delaying their toxicity for endothelial cells. However, taking into consideration MMP ability to degrade basement membrane components, these protective effects might also undesirably compromise blood brain barrier integrity and precipitate a hemorrhagic phenotype.
authors.affiliation: Neurovascular Research Laboratory, Institut de Recerca, Hospital Vall d'Hebron, Universitat Autònoma de Barcelona, 08035 Barcelona, Spain.
authors.forename: Mar
authors.initials: M
authors.lastname: Hernandez-Guillamon
authors.affiliation: Department of Pathology, New York University School of Medicine, New York, New York 10016.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Mawhirt
authors.affiliation: Department of Pathology, New York University School of Medicine, New York, New York 10016.
authors.forename: Silvia
authors.initials: S
authors.lastname: Fossati
authors.affiliation: Department of Pharmacology, , New York University School of Medicine, New York, New York 10016; Kimmel Center for Biology and Medicine at the Skirball Institute, New York University School of Medicine, New York, New York 10016.
authors.forename: Steven
authors.initials: S
authors.lastname: Blais
authors.affiliation: Neurovascular Research Laboratory, Institut de Recerca, Hospital Vall d'Hebron, Universitat Autònoma de Barcelona, 08035 Barcelona, Spain.
authors.forename: Mireia
authors.initials: M
authors.lastname: Pares
authors.affiliation: Neurovascular Research Laboratory, Institut de Recerca, Hospital Vall d'Hebron, Universitat Autònoma de Barcelona, 08035 Barcelona, Spain.
authors.forename: Anna
authors.initials: A
authors.lastname: Penalba
authors.affiliation: Neurovascular Unit, Department of Neurology, Hospital Universitari Vall d'Hebron, Universitat Autònoma de Barcelona, 08035 Barcelona, Spain.
authors.forename: Merce
authors.initials: M
authors.lastname: Boada
authors.affiliation: Institut COCHIN, INSERM, 75014 Paris, France.
authors.forename: Pierre-Olivier
authors.initials: PO
authors.lastname: Couraud
authors.affiliation: Department of Pharmacology, , New York University School of Medicine, New York, New York 10016; Kimmel Center for Biology and Medicine at the Skirball Institute, New York University School of Medicine, New York, New York 10016.
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Neubert
authors.affiliation: Neurovascular Research Laboratory, Institut de Recerca, Hospital Vall d'Hebron, Universitat Autònoma de Barcelona, 08035 Barcelona, Spain; Neurovascular Unit, Department of Neurology, Hospital Universitari Vall d'Hebron, Universitat Autònoma de Barcelona, 08035 Barcelona, Spain.
authors.forename: Joan
authors.initials: J
authors.lastname: Montaner
authors.affiliation: Department of Pathology, New York University School of Medicine, New York, New York 10016; Department of Psychiatry, New York University School of Medicine, New York, New York 10016.
authors.forename: Jorge
authors.initials: J
authors.lastname: Ghiso
authors.affiliation: Department of Pathology, New York University School of Medicine, New York, New York 10016. Electronic address: agueda.rostagno@nyumc.org.
authors.forename: Agueda
authors.initials: A
authors.lastname: Rostagno
mesh_terms: D019943:Amino Acid Substitution; D016229:Amyloid beta-Peptides / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D001812:Blood-Brain Barrier / Q000187:drug effects / Q000201:enzymology; D001921:Brain / Q000201:enzymology* / Q000473:pathology; D042783:Endothelial Cells / Q000201:enzymology* / Q000473:pathology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000235:genetics; D020868:Gene Silencing; D006801:Humans; D020300:Intracranial Hemorrhages / Q000201:enzymology / Q000378:metabolism / Q000473:pathology; D020778:Matrix Metalloproteinase 2 / Q000096:biosynthesis* / Q000235:genetics; D020125:Mutation, Missense; D010455:Peptides / Q000235:genetics / Q000378:metabolism / Q000493:pharmacokinetics*
chemical_list: D016229:Amyloid beta-Peptides; D010455:Peptides; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2