title: AR-12 Inhibits Multiple Chaperones Concomitant With Stimulating Autophagosome Formation Collectively Preventing Virus Replication.
abstract: We have recently demonstrated that AR-12 (OSU-03012) reduces the function and ATPase activities of multiple HSP90 and HSP70 family chaperones. Combined knock down of chaperones or AR-12 treatment acted to reduce the expression of virus receptors and essential glucosidase proteins. Combined knock down of chaperones or AR-12 treatment inactivated mTOR and elevated ATG13 S318 phosphorylation concomitant with inducing an endoplasmic reticulum stress response that in an eIF2α-dependent fashion increased Beclin1 and LC3 expression and autophagosome formation. Over-expression of chaperones prevented the reduction in receptor/glucosidase expression, mTOR inactivation, the ER stress response, and autophagosome formation. AR-12 reduced the reproduction of viruses including Mumps, Influenza, Measles, Junín, Rubella, HIV (wild type and protease resistant), and Ebola, an effect replicated by knock down of multiple chaperone proteins. AR-12-stimulated the co-localization of Influenza, EBV and HIV virus proteins with LC3 in autophagosomes and reduced viral protein association with the chaperones HSP90, HSP70, and GRP78. Knock down of Beclin1 suppressed drug-induced autophagosome formation and reduced the anti-viral protection afforded by AR-12. In an animal model of hemorrhagic fever virus, a transient exposure of animals to low doses of AR-12 doubled animal survival from ∼30% to ∼60% and suppressed liver damage as measured by ATL, GGT and LDH release. Thus through inhibition of chaperone protein functions; reducing the production, stability and processing of viral proteins; and stimulating autophagosome formation/viral protein degradation, AR-12 acts as a broad-specificity anti-viral drug in vitro and in vivo. We argue future patient studies with AR-12 are warranted. J. Cell. Physiol. 231: 2286-2302, 2016. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Department of Biochemistry and Molecular Biology, Virginia Commonwealth University, Richmond, Virginia.
authors.forename: Laurence
authors.initials: L
authors.lastname: Booth
authors.affiliation: Department of Biochemistry and Molecular Biology, Virginia Commonwealth University, Richmond, Virginia.
authors.forename: Jane L
authors.initials: JL
authors.lastname: Roberts
authors.affiliation: School of Biological Sciences and Illawarra Health and Medical Research Institute, University of Wollongong, New South Wales, Australia.
authors.forename: Heath
authors.initials: H
authors.lastname: Ecroyd
authors.affiliation: Molecular and Translational Science, United States Army Medical Research Institute of Infectious Diseases (USAMRIID), Fort Detrick, Frederick, Maryland.
authors.forename: Sarah R
authors.initials: SR
authors.lastname: Tritsch
authors.affiliation: Molecular and Translational Science, United States Army Medical Research Institute of Infectious Diseases (USAMRIID), Fort Detrick, Frederick, Maryland.
authors.forename: Sina
authors.initials: S
authors.lastname: Bavari
authors.affiliation: Molecular and Translational Science, United States Army Medical Research Institute of Infectious Diseases (USAMRIID), Fort Detrick, Frederick, Maryland.
authors.forename: St Patrick
authors.initials: SP
authors.lastname: Reid
authors.affiliation: Arno Therapeutics, Flemington, New Jersey.
authors.forename: Stefan
authors.initials: S
authors.lastname: Proniuk
authors.affiliation: Arno Therapeutics, Flemington, New Jersey.
authors.forename: Alexander
authors.initials: A
authors.lastname: Zukiwski
authors.affiliation: Department of Otolaryngology, University of Arizona Ear Institute, Tucson, Arizona.
authors.forename: Abraham
authors.initials: A
authors.lastname: Jacob
authors.affiliation: FCEN-UBA, Ciudad Universitaria, Pabellón 2 Piso 4, lab QB-17, Buenos Aires, Argentina.
authors.forename: Claudia S
authors.initials: CS
authors.lastname: Sepúlveda
authors.affiliation: FCEN-UBA, Ciudad Universitaria, Pabellón 2 Piso 4, lab QB-17, Buenos Aires, Argentina.
authors.forename: Federico
authors.initials: F
authors.lastname: Giovannoni
authors.affiliation: FCEN-UBA, Ciudad Universitaria, Pabellón 2 Piso 4, lab QB-17, Buenos Aires, Argentina.
authors.forename: Cybele C
authors.initials: CC
authors.lastname: García
authors.affiliation: FCEN-UBA, Ciudad Universitaria, Pabellón 2 Piso 4, lab QB-17, Buenos Aires, Argentina.
authors.forename: Elsa
authors.initials: E
authors.lastname: Damonte
authors.affiliation: Institute of Biomedicine and CIBEREhd, University of León, León, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: González-Gallego
authors.affiliation: Institute of Biomedicine and CIBEREhd, University of León, León, Spain.
authors.forename: María J
authors.initials: MJ
authors.lastname: Tuñón
authors.affiliation: Department of Biochemistry and Molecular Biology, Virginia Commonwealth University, Richmond, Virginia.
authors.forename: Paul
authors.initials: P
authors.lastname: Dent
mesh_terms: D000071182:Autophagosomes / Q000187:drug effects*; D001343:Autophagy / Q000187:drug effects*; D002460:Cell Line; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism; D000091342:Endoplasmic Reticulum Chaperone BiP; D059865:Endoplasmic Reticulum Stress / Q000187:drug effects / Q000502:physiology; D006801:Humans; D018832:Molecular Chaperones / Q000378:metabolism*; D011720:Pyrazoles / Q000494:pharmacology*; D013449:Sulfonamides / Q000494:pharmacology*; D014779:Virus Replication / Q000187:drug effects* / Q000502:physiology
chemical_list: D000091342:Endoplasmic Reticulum Chaperone BiP; C000717247:HSPA5 protein, human; D018832:Molecular Chaperones; C500894:OSU 03012; D011720:Pyrazoles; D013449:Sulfonamides