乙型肝炎病毒X蛋白对HBV复制相关微染色体重塑的影响的开题报告.docx

乙型肝炎病毒X蛋白对HBV复制相关微染色体重塑的影响的开题报告

Introduction:

HBV(hepatitisBvirus)isamajorcauseofliverdisease,affectingmillionsofpeopleworldwide.ThereplicationofHBViscloselylinkedtothechromatinstructureofthehostcell.TheXprotein(HBx)ofHBVisamultifunctionalproteinthatplaysacriticalroleinthereplicationofHBVandinthedevelopmentoflivercancer.However,themechanismsbywhichHBxaltersthechromatinstructureofhostcellstofacilitateHBVreplicationarenotwellunderstood.

Objectives:

TheobjectiveofthisstudyistoinvestigatetheeffectsofHBxonchromatinremodelinginHBV-infectedcells.Specifically,weaimto:

1.CharacterizethechangesintheepigeneticlandscapeofHBV-infectedcellsinthepresenceofHBx.

2.DeterminetheimpactofHBxontheorganizationandaccessibilityofHBVcccDNAinthenucleus.

3.InvestigatetheroleofchromatinmodifyingenzymesinmediatingtheeffectsofHBxonchromatinstructure.

Methods:

TheproposedstudywillinvolvetheuseofhumanhepatomacelllinesinfectedwithHBV.ThecellswillbetransfectedwithHBx-expressingvectorstooverexpressHBxprotein.Thechromatinremodelinginthesecellswillbeinvestigatedusingchromatinimmunoprecipitation(ChIP)assaystoanalyzethehistonemodificationsandDNAaccessibility.Additionally,thelocalizationandaccessibilityofHBVcccDNAinthenucleuswillbeassessedusingfluorescenceinsituhybridization(FISH)andChIPassays.Theinvolvementofchromatinmodifyingenzymeswillbeinvestigatedbyusinginhibitorsofspecificenzymes.

Expectedoutcomes:

WepredictthatHBxproteinofHBVwillalterthechromatinstructureofhostcellstopromoteHBVreplicationbyincreasingtheaccessibilityofHBVcccDNA.Wealsoanticipatethatthechangesinthechromatinstructurewillbeassociatedwithalterationsinvariousepigeneticmodifications.ThestudyisexpectedtoprovideinsightsintothemechanismsunderlyingHBVreplicati