Adenoviral Vectors Stimulate Glucagon Transcription in Human Mesenchymal Stem Cells Expressing Pancreatic Transcription Factors 英文参考文献.docVIP

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Adenoviral Vectors Stimulate Glucagon Transcription in Human Mesenchymal Stem Cells Expressing Pancreatic Transcription Factors 英文参考文献.doc

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Adenoviral Vectors Stimulate Glucagon Transcription in Human Mesenchymal Stem Cells Expressing Pancreatic Transcription Factors 英文参考文献

AdenoviralVectorsStimulateGlucagonTranscriptionin HumanMesenchymalStemCellsExpressingPancreatic TranscriptionFactors ArnaudZaldumbide1*,Franc?oiseCarlotti2,ManuelA.Gonc?alves1,ShoshanKnaa¨n-Shanzer1, SteveJ.Cramer1,BartO.Roep3,EmmanuelJ.H.J.Wiertz4,RobC.Hoeben1 1Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands, 2Department of Nephrology, Leiden University Medical Center, Leiden, The Netherlands, 3Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, Leiden, The Netherlands, 4Department of MedicalMicrobiology,UniversityMedicalCenterUtrecht,Utrecht,TheNetherlands Abstract Viralgenecarriersarebeingwidelyusedasgenetransfersystemsin(trans)differentiationandreprogrammingstrategies. Forcedexpressionofkeyregulatorsofpancreaticdifferentiationinstemcells,livercells,pancreaticductcells,orcellsfrom theexocrinepancreas,canleadtotheinitiationofendocrinepancreaticdifferentiation.Whileseveralviralvectorsystems havebeenemployedinsuchstudies,theresultsreportedwithadenovirusvectorshavebeenthemostpromisinginvitro andinvivo.Inthisstudy,weexaminedwhethertheviralvectorsystemitselfcouldimpactthedifferentiationcapacityof human bone-marrow derived mesenchymal stem cells (hMSCs) toward the endocrine lineage. Lentivirus-mediated expressionofPdx-1,Ngn-3,andMaf-Aaloneorincombinationdoesnotleadtorobustexpressionofanyoftheendocrine hormones (i.e. insulin, glucagon and somatostatin) in hMSCs. Remarkably, subsequent transduction of these genetically modifiedcellswithanirrelevantearlyregion1(E1)-deletedadenoviralvectorpotentiatesthedifferentiationstimulusand promotes glucagon gene expression in hMSCs by affecting the chromatin structure. This adenovirus stimulation was observed upon infection with an E1-deleted adenovirus vector, but not after exposure to helper-dependent adenovirus vectors,pointingattheinvolvementofgenesretainedintheE1-deletedadenovirusvectorinthisphenomenon.Lentivirus mediatedexpressionoftheadenoviru