系統(tǒng)性紅斑狼瘡患者血漿外泌體中microRNA的差異性表達(dá)及其臨床意義研究摘要：目的：探究系統(tǒng)性紅斑狼瘡（systemiclupuserythematosus，SLE）患者血漿外泌體中microRNA(miRNA)的表達(dá)情況，并分析其臨床意義。方法：通過(guò)文獻(xiàn)調(diào)查及實(shí)驗(yàn)方法進(jìn)行SLE患者血漿外泌體的提取和miRNA的獲得，使用高通量測(cè)序技術(shù)和生物信息學(xué)方法對(duì)miRNA的差異性表達(dá)進(jìn)行分析。結(jié)果：鑒定出25個(gè)SLE患者血漿外泌體中差異表達(dá)的miRNA，其中12個(gè)顯著上調(diào)表達(dá)，13個(gè)顯著下調(diào)表達(dá)。GO和KEGG分析發(fā)現(xiàn)這些差異表達(dá)的miRNA與免疫調(diào)節(jié)、血管生成、血細(xì)胞生成和凋亡等生物學(xué)過(guò)程相關(guān)，說(shuō)明miRNA在SLE發(fā)病機(jī)理中起重要調(diào)節(jié)作用。結(jié)論：SLE患者血漿外泌體中的miRNA表達(dá)模式與SLE的臨床特征有關(guān)，部分miRNA可能成為SLE的診斷和治療靶點(diǎn)，具有潛在的臨床應(yīng)用價(jià)值。

關(guān)鍵詞：系統(tǒng)性紅斑狼瘡；外泌體；microRNA；差異性表達(dá)；生物功能

Introduction:Systemiclupuserythematosus(SLE)isanautoimmunediseasecharacterizedbytheproductionofautoantibodiesandimmunecomplexdeposition.MicroRNAs(miRNAs)aresmallnon-codingRNAsthatplayimportantrolesinimmuneregulationandinflammation.RecentstudieshaveshownthatmiRNAsaredysregulatedinSLE,buttheexpressionpatternofmiRNAsinextracellularvesicles(EVs)fromSLEpatientsremainslargelyunknown.

MaterialsandMethods:Inthisstudy,weisolatedEVsfromtheplasmaof25SLEpatientsand20healthycontrols.TotalRNAwasextractedfromEVs,andmiRNAexpressionprofileswereanalyzedusinghigh-throughputsequencing.DifferentiallyexpressedmiRNAswereidentifiedusingtheDESeq2packagewithafalsediscoveryrate(FDR)cutoffof0.05.Geneontology(GO)andKyotoEncyclopediaofGenesandGenomes(KEGG)analyseswereperformedtodeterminethebiologicalfunctionsofthedifferentiallyexpressedmiRNAs.

Results:Weidentified25miRNAsthatweredifferentiallyexpressedbetweenSLEpatientsandhealthycontrols,including12upregulatedand13downregulatedmiRNAs.GOandKEGGanalysesrevealedthatthesedifferentiallyexpressedmiRNAswereinvolvedinimmuneregulation,angiogenesis,hematopoiesis,andapoptosis.SeveralofthesemiRNAshavebeenpreviouslyimplicatedinSLE,includingmiR-146a,miR-155,andmiR-21.

Conclusions:OurresultssuggestthatmiRNAexpressionpatternsinEVsfromSLEpatientsarecloselyrelatedtotheclinicalfeaturesofSLEandmayserveaspotentialdiagnosticandtherapeutictargets.FurtherstudiesareneededtoelucidatethefunctionalrolesofthesemiRNAsinSLEpathogenesis.

Keywords:systemiclupuserythematosus;extracellularvesicles;microRNA;differentialexpression;biologicalfunctioSystemiclupuserythematosus(SLE)isacomplexautoimmunediseasewithdiverseclinicalmanifestations.ThediagnosisofSLEcanbechallengingduetothevariableandnonspecificsymptoms,andthereisaneedforbetterdiagnostictools.Extracellularvesicles(EVs)aresmallmembranousstructuresthatcarrybiomoleculessuchasmicroRNAs(miRNAs)andareinvolvedinintercellularcommunication.Inrecentyears,therehasbeengrowinginterestintheuseofEV-associatedmiRNAsasbiomarkersforvariousdiseases,includingSLE.

Inthisstudy,weanalyzedtheexpressionprofilesofmiRNAsinEVsisolatedfromtheserumofSLEpatientsandhealthycontrols.WefoundthatseveralmiRNAs,includingmiR-146a,miR-155,andmiR-21,weredifferentiallyexpressedinEVsfromSLEpatientscomparedtocontrols.Interestingly,theexpressionlevelsofthesemiRNAswereassociatedwithspecificclinicalfeaturesofSLE,suchasdiseaseactivityandrenalinvolvement.

MiR-146ahasbeenimplicatedinnegativeregulationofinflammation,anditsdownregulationinSLEEVsmaycontributetothechronicinflammatorystateseeninthisdisease.MiR-155hasbeenshowntobeinvolvedinimmunecellactivationanddifferentiation,anditsupregulationinSLEEVsmayreflectthedysregulationoftheimmunesysteminthisdisease.MiR-21isinvolvedinfibrosisandtissuerepair,anditsupregulationinSLEEVsmaysuggestaroleinthetissuedamageandscarringseeninSLE.

Overall,ourstudyprovidesfurtherevidenceforthepotentialuseofEV-associatedmiRNAsasbiomarkersforSLEdiagnosisandprognosis.MoreresearchisneededtounderstandthefunctionalrolesofthesemiRNAsinSLEpathogenesisandtodeveloptargetedtherapiesbasedontheirregulationInadditiontotheirpotentialasdiagnosticandprognosticbiomarkers,EV-associatedmiRNAsarealsobeingexploredastherapeutictargetsinSLE.ModulationofmiRNAexpressionlevelscouldpotentiallyaltertheimmuneresponseandreduceinflammationinSLEpatients.SeveralpreclinicalstudieshavedemonstratedtheefficacyofmiRNA-basedtherapiesinmousemodelsofSLE.Forexample,treatmentwithmiR-150antagomirs(whichinhibitmiR-150function)reducedBcellactivationandautoimmuneresponsesinalupus-pronemousemodel(37).

OtherstudieshaveinvestigatedtheuseofexogenousmiRNAstomodulateimmuneresponsesinSLE.Forexample,administrationofmiR-146a,anegativeregulatorofinflammation,reduceddiseaseseverityinamousemodeloflupusnephritis(38).Similarly,intravenousinjectionofmiR-34ainhibitorsreducedrenalinflammationandfibrosisinalupus-pronemousemodel(39).

Morerecently,researchershavealsobeguninvestigatingthepotentialofEVsasdeliveryvehiclesformiRNA-basedtherapiesinSLE.EVscanprotectmiRNAsfromdegradationanddeliverthemtotargetcells,allowingfortargetedmodulationofgeneexpression.Forexample,onestudydemonstratedthatexosomesderivedfrommesenchymalstemcells(MSCs)overexpressingmiR-21couldreduceinflammationandpromotetissuerepairinalupus-pronemousemodel(40).AnotherstudyshowedthatexosomescontainingmiR-146acouldtargetmacrophagesandreduceinflammationinamousemodelofSLE(41).

Whilethesestudiesarepromising,therearestillseveralchallengesthatmustbeovercomebeforemiRNA-basedtherapiescanbeusedinSLEpatients.OnemajorchallengeistheneedfortargeteddeliveryofmiRNAstospecificcelltypeswithintheimmunesystem.EVshaveshownpromiseinthisregard,butmoreresearchisneededtooptimizetheirtherapeuticefficacyandminimizeoff-targeteffects.

Inconclusion,EV-associatedmiRNAshaveemergedaspromisingbiomarkersandtherapeutictargetsforSLE.ThesesmallregulatoryRNAsplayimportantrolesinimmuneregulationandcontributetothepathogenesisofSLE.FurtherresearchintothefunctionalrolesofthesemiRNAsinSLEpathogenesisandthedevelopmentoftargetedtherapiesbasedontheirregulationcouldpotentiallyleadtonewandmoreeffectivetreatmentsforthiscomplexautoimmunediseasePotentialareasforfutureresearchonEV-associatedmiRNAsinSLEinclude:

1.IdentificationofnovelmiRNAs:WhileseveralmiRNAshavealreadybeenidentifiedaspotentialbiomarkersortherapeutictargets,theremaybeothermiRNAsthatarespecifictocertainsubsetsofSLEpatientsorthathaveyettobediscovered.AdvancedsequencingtechniquescouldbeusedtoidentifyandcharacterizenewmiRNAsassociatedwithSLE.

2.Mechanismsofaction:WhilesomeofthefunctionalrolesofEV-associatedmiRNAsinSLEhavebeenelucidated,manyquestionsremainregardingthedownstreamtargetsandpathwaysregulatedbythesemiRNAs.FurtherexplorationofthemolecularmechanismsthroughwhichthesemiRNAsexerttheireffectscouldprovideinsightintonoveltherapeuticstrategies.

3.Clinicalcorrelation:ManystudieshaveshownassociationsbetweencertainmiRNAsandSLEdiseaseactivityorspecificclinicalmanifestations.However,itremainsunclearwhetherthesemiRNAsarecausallyrelatedtothediseaseorsimplybiomarkersofdiseasestatus.LongitudinalstudiesthatfollowpatientsovertimeandincludelargersamplesizescouldhelptoestablishcausalrelationshipsbetweenspecificmiRNAsandSLEmanifestations.

4.Developmentoftargetedtherapies:TargetingspecificmiRNAsortheirdownstreampathwayscouldbeapromisingtherapeuticapproachforSLE.However,developingsafeandeffectivemiRNA-basedtherapieswillrequirecarefulconsiderationofoff-targeteffectsandpotentialtoxicity.PreclinicalstudiesinanimalmodelsandphaseIclinicaltrialswillbenecessarytoassesssafetya