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1、天津醫(yī)科大學碩士畢業(yè)論文AbstractObjective:Duchennemusculardystrophy(DMD)isanX-linked,lethalneuromusculardisordercausedbyframe-disruptingmutationsinthedystrophingene.Currentlythereisnotreatmentavailableinclinic.Antisenseoligonucleotidemediatedexon-skippingtherapeuticsisoneofthemostpromisingapp
2、roachesfortreatingDMD.However,recentphaseIIIclinicaltrialfailedtomeettheprimaryendpoints,whichislikelyattributedtotheinsufficientsystemicdeliveryasdemonstratedinpre-clinicalstudies.Thisfurtherunderlinedtheimportantanceofdevelopingsafe,effectivedeliverytechnologies,compatiblewithth
3、eAOsdevelopment.Previously,wescreenedanumberofdifferentformulationsinmdxmiceintramuscularlyandidentifiedafewtopcandidatesincludingF5,whichcansignificantlyenhancethepotencyofAOs.Basedonpreviousstudies,wetriedtoexplorethepotentialofF5inmdxmicesystemicallybycomparingF5withotherfourto
4、pcandidatesintheabilitytoenhancetheactivityofdifferentAOsinmdxmiceintramuscularly.Furthermore,weinvestigatedthesystemiceffectofF5onaugmentingtheactivityofAOsandthepotentialdrug-relatedtoxicitiesinmdxmiceviadifferentdosingregimens.OurstudywillprovideanewavenueandstrategyforAOsystem
5、icdeliveryforDMDandserveasacornerstoneforclinicaltranslationofF5.SecondpartofourstudywastoexploreotherpotentialDMDmousemodelsforacceleratingthedrugdevelopmentforDMDgiventheaccessibilityandhighmaintainencecostofmdxmice.Methods:1.Basedonpreviouswork,wechosefivetopcandidateformulatio
6、nsincludingF4,F5,F9,F10andF12forfurtherevaluationinmdxmice.Fivecandidateswereco-injectedwith5μgMOEAOsintotibialisanterior(TA)musclesofmdxmiceandTAmuscleswereharvestedtwoweekspost-injection.Immunohistochemistry,RT-PCRandWesternblotwereusedforexaminingtheexpressionanddistributionofd
7、ystrophin-positivefibersandthelevelofexon-skippinganddystrophinrestoration.2.Fivecandidateformulationswereco-injectedwith5μgpeptidenucleicacid(PNA)AOsintoTAmusclesofmdxmiceandTAmuscleswereharvestedtwoweekspost-injection.Immunohistochemistry,RT-PCRandWesternblotwereusedforexamining
8、theexpressionanddistributionofdys