MicroRNA122,RegulatedbyGRLH2,ProtectsLiversofMiceandPatientsfromEthanol-inducedLiverDiseaseAbhishekSatishchandran1[GraduateStudent],AdityaAmbade1[PostDoctoralAssociate],SitaraRao1[ResearchTechnician],Ying-ChaoHsueh1[GraduateStudent],ArvinIracheta-Vellve1[GraduateStudent],DavidTornai1[GraduateStudent],PatrickLowe1[GraduateStudent],BenedekGyongyosi1[PostDoctoralAssociate],JiaLi2,3[researchtechnician],DonnaCatalano1[ResearchAssistant],LiZhong2,3[AssociateResearchProfessor],KarenKodys1[LabManager],JunXie2,4[Instructor],ShashiBala1[Instructor],GuangpingGao2,4[Professor],andGyongyiSzabo1,*[Professor]1Dept.ofMedicine,UniversityofMassachusettsMedicalSchool2GeneTherapyCenter,UniversityofMassachusettsMedicalSchool3Dept.ofPediatrics,UniversityofMassachusettsMedicalSchool4Dept.ofMicrobiologyandPhysiologicalSystems,UniversityofMassachusettsMedicalSchoolAbstractBackground&Aims—Chronic,excessivealcoholconsumptionleadstoalcoholicliverdisease(ALD)characterizedbysteatosis,inflammation,andeventuallycirrhosis.ThehepatocytespecificmicroRNA122(MIR122)regulateshepatocytedifferentiationandmetabolism.Weinvestigatedwhetheranalcohol-induceddecreaseinlevelofMIR122contributestodevelopmentofALD.Methods—Weobtainedliversamplesfrom12patientswithALDandcirrhosisand9healthyindividuals(controls)andanalyzedthembyhistologyandimmunohistochemistry.C57Bl/6micewereplacedonaLieber-DeCarliliquiddiet,inwhichtheywerefedethanolfor8weeks,asamodelofALD,oracontroldiet.ThesemicewerealsogiveninjectionsofCCl4,toincreaseliverfibrosis,for8weeks.Onday28,micewithethanol-inducedliverdiseaseandadvancedfibrosis,andcontrols,weregiveninjectionsofrecombinantadeno-associatedvirus8vectorthatexpressedtheprimarymiR-122transcript(pri-MIR122,tooverexpressMIR122inhepatocytes)orvector(control).Twoweeksbeforeethanolfeeding,somemiceweregiveninjectionsofavectorthatexpressedananti-MIR122,toknockdownitsexpression.Serumandlivertissueswerecollected;hepatocytesandlivermononuclearcellswereanalyzedbyhistology,immunoblots,andconfocalmicroscop...
