title: Proteomic analysis of ischemia-reperfusion injury upon human liver transplantation reveals the protective role of IQGAP1.
abstract: Ischemia-reperfusion injury (IRI) represents a major determinant of liver transplantation. IRI-induced graft dysfunction is related to biliary damage, partly due to a loss of bile canaliculi (BC) integrity associated with a dramatic remodeling of actin cytoskeleton. However, the molecular mechanisms associated with these events remain poorly characterized. Using liver biopsies collected during the early phases of organ procurement (ischemia) and transplantation (reperfusion), we characterized the global patterns of expression and phosphorylation of cytoskeleton-related proteins during hepatic IRI. This targeted functional proteomic approach, which combined protein expression pattern profiling and phosphoprotein enrichment followed by mass spectrometry analysis, allowed us to identify IQGAP1, a Cdc42/Rac1 effector, as a potential regulator of actin cytoskeleton remodeling and maintenance of BC integrity. Cell fractionation and immunohistochemistry revealed that IQGAP1 expression and localization were affected upon IRI and related to actin reorganization. Furthermore using an IRI model in human hepatoma cells, we demonstrated that IQGAP1 silencing decreased the basal level of actin polymerization at BC periphery, reflecting a defect in BC structure coincident with reduced cellular resistance to IRI. In summary, this study uncovered new mechanistic insights into the global regulation of IRI-induced cytoskeleton remodeling and led to the identification of IQGAP1 as a regulator of BC structure. IQGAP1 therefore represents a potential target for the design of new organ preservation strategies to improve transplantation outcome.
authors.affiliation: Organelle Signalling Laboratory, Hepato-Biliary and Transplant Research Group, Department of Surgery, McGill University, Montreal, Quebec, Canada.
authors.forename: Anouk
authors.initials: A
authors.lastname: Emadali
authors.forename: Béatrice
authors.initials: B
authors.lastname: Muscatelli-Groux
authors.forename: Frédéric
authors.initials: F
authors.lastname: Delom
authors.forename: Sarah
authors.initials: S
authors.lastname: Jenna
authors.forename: Daniel
authors.initials: D
authors.lastname: Boismenu
authors.forename: David B
authors.initials: DB
authors.lastname: Sacks
authors.forename: Peter P
authors.initials: PP
authors.lastname: Metrakos
authors.forename: Eric
authors.initials: E
authors.lastname: Chevet
mesh_terms: D000199:Actins / Q000378:metabolism; D001648:Bile Canaliculi / Q000033:anatomy & histology; D001706:Biopsy; D016764:Cell Polarity; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D005708:Gallium / Q000032:analysis / Q000378:metabolism; D022781:Hepatocytes / Q000378:metabolism; D006801:Humans; D007365:Intercellular Junctions; D008099:Liver / Q000601:surgery; D016031:Liver Transplantation / Q000502:physiology*; D011108:Polymers / Q000378:metabolism; D040901:Proteomics; D015427:Reperfusion Injury / Q000209:etiology* / Q000534:rehabilitation; D014018:Tissue Distribution; D014162:Transfection; D015032:Zinc / Q000032:analysis / Q000378:metabolism; D020703:ras GTPase-Activating Proteins / Q000378:metabolism / Q000502:physiology*
chemical_list: D000199:Actins; D003598:Cytoskeletal Proteins; C101203:IQ motif containing GTPase activating protein 1; C101002:IQGAP2 protein, human; D011108:Polymers; D020703:ras GTPase-Activating Proteins; D005708:Gallium; D015032:Zinc