title: AAV-mediated in vivo functional selection of tissue-protective factors against ischaemia.
abstract: Functional screening of expression libraries in vivo would offer the possibility of identifying novel biotherapeutics without a priori knowledge of their biochemical function. Here we describe a procedure for the functional selection of tissue-protective factors based on the in vivo delivery of arrayed cDNA libraries from the mouse secretome using adeno-associated virus (AAV) vectors. Application of this technique, which we call FunSel, in the context of acute ischaemia, revealed that the peptide ghrelin protects skeletal muscle and heart from ischaemic damage. When delivered to the heart using an AAV9 vector, ghrelin markedly reduces infarct size and preserves cardiac function over time. This protective activity associates with the capacity of ghrelin to sustain autophagy and remove dysfunctional mitochondria after myocardial infarction. Our findings describe an innovative tool to identify biological therapeutics and reveal a novel role of ghrelin as an inducer of myoprotective autophagy.
authors.affiliation: Molecular Medicine Laboratory, International Centre for Genetic Engineering and Biotechnology (ICGEB), 34149 Trieste, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Ruozi
authors.affiliation: Molecular Medicine Laboratory, International Centre for Genetic Engineering and Biotechnology (ICGEB), 34149 Trieste, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Bortolotti
authors.affiliation: Molecular Medicine Laboratory, International Centre for Genetic Engineering and Biotechnology (ICGEB), 34149 Trieste, Italy.
authors.forename: Antonella
authors.initials: A
authors.lastname: Falcione
authors.affiliation: 1] Molecular Medicine Laboratory, International Centre for Genetic Engineering and Biotechnology (ICGEB), 34149 Trieste, Italy [2] Department of Medical, Surgical and Health Sciences, University of Trieste, 34127 Trieste, Italy.
authors.forename: Matteo
authors.initials: M
authors.lastname: Dal Ferro
authors.affiliation: Molecular Medicine Laboratory, International Centre for Genetic Engineering and Biotechnology (ICGEB), 34149 Trieste, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Ukovich
authors.affiliation: Molecular Medicine Laboratory, International Centre for Genetic Engineering and Biotechnology (ICGEB), 34149 Trieste, Italy.
authors.forename: Antero
authors.initials: A
authors.lastname: Macedo
authors.affiliation: Molecular Medicine Laboratory, International Centre for Genetic Engineering and Biotechnology (ICGEB), 34149 Trieste, Italy.
authors.forename: Lorena
authors.initials: L
authors.lastname: Zentilin
authors.affiliation: Department of Translational Medicine, University of Piemonte Orientale 'A. Avogadro', 28100 Novara, Italy.
authors.forename: Nicoletta
authors.identifier: 000000023848611X
authors.initials: N
authors.lastname: Filigheddu
authors.affiliation: Department of Medical, Surgical and Health Sciences, University of Trieste, 34127 Trieste, Italy.
authors.forename: Gianluca
authors.identifier: 0000000204385003
authors.initials: G
authors.lastname: Gortan Cappellari
authors.affiliation: Department of Medical, Surgical and Health Sciences, University of Trieste, 34127 Trieste, Italy.
authors.forename: Giovanna
authors.identifier: 0000000230662780
authors.initials: G
authors.lastname: Baldini
authors.affiliation: Department of Medical, Surgical and Health Sciences, University of Trieste, 34127 Trieste, Italy.
authors.forename: Marina
authors.initials: M
authors.lastname: Zweyer
authors.affiliation: Department of Medical, Surgical and Health Sciences, University of Trieste, 34127 Trieste, Italy.
authors.forename: Rocco
authors.initials: R
authors.lastname: Barazzoni
authors.affiliation: Department of Translational Medicine, University of Piemonte Orientale 'A. Avogadro', 28100 Novara, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Graziani
authors.affiliation: 1] Molecular Medicine Laboratory, International Centre for Genetic Engineering and Biotechnology (ICGEB), 34149 Trieste, Italy [2] Department of Medical, Surgical and Health Sciences, University of Trieste, 34127 Trieste, Italy.
authors.forename: Serena
authors.initials: S
authors.lastname: Zacchigna
authors.affiliation: 1] Molecular Medicine Laboratory, International Centre for Genetic Engineering and Biotechnology (ICGEB), 34149 Trieste, Italy [2] Department of Medical, Surgical and Health Sciences, University of Trieste, 34127 Trieste, Italy.
authors.forename: Mauro
authors.initials: M
authors.lastname: Giacca
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics*; D001343:Autophagy / Q000235:genetics*; D015153:Blotting, Western; D000229:Dependovirus; D004317:Doxorubicin / Q000494:pharmacology; D020869:Gene Expression Profiling; D015723:Gene Library; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D054439:Ghrelin / Q000235:genetics*; D006614:Hindlimb / Q000098:blood supply; D007124:Immunoenzyme Techniques; D020287:In Situ Nick-End Labeling; D007511:Ischemia / Q000235:genetics / Q000378:metabolism; D051379:Mice; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D008929:Mitochondria, Heart / Q000378:metabolism* / Q000648:ultrastructure; D018482:Muscle, Skeletal / Q000098:blood supply; D009203:Myocardial Infarction / Q000000981:diagnostic imaging / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D017202:Myocardial Ischemia / Q000235:genetics / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D014463:Ultrasonography; D018487:Ventricular Dysfunction, Left / Q000000981:diagnostic imaging / Q000235:genetics / Q000378:metabolism / Q000473:pathology
chemical_list: D000903:Antibiotics, Antineoplastic; D054439:Ghrelin; D004317:Doxorubicin