心肌素相关转录因子调控米色脂肪细胞的产生

心肌素相关转录因子调控米色脂肪细胞的产生第1页，课件共34页，创作于2023年2月Whiteadiposetissue

storesexcessenergyintheformoftriglycerides.BrownadiposetissuedissipatesenergyasheatviatheuniqueexpressionofthemitochondrialuncouplingproteinUCP1.Thedevelopmentaloriginofbeigeadipocytesisunclear.TheydefinatedaBMP7-ROCKsignalingaxisregulatingbeigeadipocyteformationviacontroloftheG-actin-regulatedtranscriptionalcoactivatormyocardin-relatedtranscriptionfactorA,MRTFA.Summary第2页，课件共34页，创作于2023年2月WhiteadiposetissuefromMRTFA–/–micecontainsmoremultilocularadipocytesandexpressesenhancedlevelsofbrown-selectiveproteins,includingUCP1.MRTFA–/–micealsoshowimprovedmetabolicprofilesandprotectionfromdiet-inducedobesityandinsulinresistance.Thestudyhenceunravelsacentralpathwaydrivingthedevelopmentofphysiologicallyfunctionalbeigeadipocytes.Summary第3页，课件共34页，创作于2023年2月1Introduction2Result3DiscussionCONTENTS第4页，课件共34页，创作于2023年2月IntroductionWhite&BrownadipocyteWhiteadipocytesstoreslipidsinuniloculardroplets,incomparison,brownadipocytescontainmultilocularlipiddropletsandnumerousmitochondria.Whiteadipocytesstoreexcessenergyintheformoftriglycerides,brownadipocytesarethermogeniccellsthat,whenactivatedbyappropriatestimuli,dissipateenergyasheat.UCP1UCP1isabrownadiposetissue(BAT)-specificmitochondrialuncouplingprotein,andtheactivityofitrelatedtoenergyrelease.第5页，课件共34页，创作于2023年2月IntroductionBeigeadipocyteUCP1+adipocytesdesignatedasbeigeadipocyte.Currenthypothesesproposethattheyarisefromthe

conversionofexistingwhiteadipocytesand/orthedifferentiationofWATresidentprogenitorcells.Brownandbeigeadipocytesinvitropossessadistinctgeneexpressionsignature.

theoriginsofprogenitorsPPARglineagetracingstudiesindicatedthatWATprogenitorsresideinthemuralcellcompartmentofadiposevasculatureandatleastapopulationofbeigecellshaveasmoothmuscle-likeorigin.Theseobservationssuggestthatwhiteandbeigeadipocytesarisefromprogenitorscloselyassociatedwiththevasculature.第6页，课件共34页，创作于2023年2月IntroductionTGFbTGF𝛃superfamilyintimatelyregulatesthedevelopmentandmaintenanceofthevasculatureviapromotingsmoothmuscledifferentiationandcoordinatingtheexpressionofSMCgenes.Italsoinhibitsadipocytedifferentiationviaitscoeffector,Smad3,whichcomplexeswithC/EBP𝛃andrepressesactivationofadipogenictargetgenes.Smad3knockout(KO)micedevelopbrown-likeadipocytesinWATdepotsandareprotectedfromobesity.BMP7Bonemorphogeneticproteins(BMPs),promoteadipocyteformation.BMP7initiatesthecommitmentofMSCstothebrownadipocytelineagebypromotingtheexpressionofthebrownadipocytefactorsPRDM16,PGC-1aandUCP1,andmitochondrialbiogenesis.Importantly,BMP7-/-miceattenuatestheformationofBAT;BMP7isalsoabletoinducetheconversionofprogenitorsisolatedfromWAT,BAT,andmuscletobrown-likeadipocyte.第7页，课件共34页，创作于2023年2月SRF/MRTFspathwayTheRho-GTPasefamilymediatesthedynamiccontrolofmonomericandfilamentousactin.MonomericG-actincanregulatethenucleus-cytoplasmshuttlingofserumresponsefactor(SRF)coregulators,myocardin-relatedtranscriptionfactors(MRTFs)and,thereby,influencetheexpressionofSRFtargetgenesincludingsmoothmuscleactin(SMA).Introduction第8页，课件共34页，创作于2023年2月1Introduction2Result3DiscussionCONTENTS第9页，课件共34页，创作于2023年2月1BMP7andTGFbmediatedistincteffectsonlineagecommitmentofMSCsthroughalterationofROCKactivity.

。2BMP7RepressesROCKActivityandAltersActinDynamicsinMSCs.3MRTFAandSRFOverexpressionPreventDifferentiationofMSCstoAdipocytes.Results4IncreasedProductionofBeigeAdipocytesinWATofMRTFA-DeficientMice.5StromalVascularCellsfromMRTFA–/–iWATUndergoBeigeAdipogenesisMoreExtensivelyThanWTSVFs.

6MRTFA-DeficientMiceAreProtectedfromDiet-InducedObesityandInsulinResistance.

。7MRTFARegulatesWhole-BodyEnergyExpenditure.第10页，课件共34页，创作于2023年2月1BMP7andTGFbmediatedistincteffectsonlineagecommitmentofMSCsthroughalterationofROCKactivity.

。2BMP7RepressesROCKActivityandAltersActinDynamicsinMSCs.3MRTFAandSRFOverexpressionPreventDifferentiationofMSCstoAdipocytes.Results4IncreasedProductionofBeigeAdipocytesinWATofMRTFA-DeficientMice.5StromalVascularCellsfromMRTFA–/–iWATUndergoBeigeAdipogenesisMoreExtensivelyThanWTSVFs.

6MRTFA-DeficientMiceAreProtectedfromDiet-InducedObesityandInsulinResistance.

。7MRTFARegulatesWhole-BodyEnergyExpenditure.第11页，课件共34页，创作于2023年2月ResultBMP7andTGFbmediatedistincteffectsonlineagecommitmentofMSCsthroughalterationofROCKactivity.

C3h/10T1/2MSCsdifferentiation.►BMP7➞MSCs➞adipocytes►TGF𝝱➞MSCs➞muscle-likecells第12页，课件共34页，创作于2023年2月ResultBMP7andTGFbmediatedistincteffectsonlineagecommitmentofMSCsthroughalterationofROCKactivity.

Bonemarrow-derivedMSCs.TGFb1alsoactivatedSMADs1/5/8indicatingthatBMP7-inducedcommitmentisnotmediatedexclusivelybyactivationoftheSMADs,butbyadditionaldownstreameffectorsoftheBMPreceptor.第13页，课件共34页，创作于2023年2月ResultBMP7andTGFbmediatedistincteffectsonlineagecommitmentofMSCsthroughalterationofROCKactivity.

DMH1:inhibitorofBMPreceptorsGremlin1:inhibitorofBMP7SMAD1/5/8↓p38MAPK↓BMP7&receptoractivatedpathwaytogether.第14页，课件共34页，创作于2023年2月1BMP7andTGFbmediatedistincteffectsonlineagecommitmentofMSCsthroughalterationofROCKactivity.

。2BMP7RepressesROCKActivityandAltersActinDynamicsinMSCs.3MRTFAandSRFOverexpressionPreventDifferentiationofMSCstoAdipocytes.Results4IncreasedProductionofBeigeAdipocytesinWATofMRTFA-DeficientMice.5StromalVascularCellsfromMRTFA–/–iWATUndergoBeigeAdipogenesisMoreExtensivelyThanWTSVFs.

6MRTFA-DeficientMiceAreProtectedfromDiet-InducedObesityandInsulinResistance.

。7MRTFARegulatesWhole-BodyEnergyExpenditure.第15页，课件共34页，创作于2023年2月ResultBMP7RepressesROCKActivityandAltersActinDynamicsinMSCs.BMP7-treatedcells:abroadenedshapeandalessprominentactinnetwork;TGFb1-treatedcells:anelongatedmorphologyandformationofactinmicrofilaments.➣BMP7➞MSCs➞ROCKactivity↓(pMYPT1↓,ROCK'starget)➣TGF𝝱➞MSCs➞ROCKactivity↑(pMYPT1↑)?

Thechangeofactindynamics⟺ROCK第16页，课件共34页，创作于2023年2月ResultBMP7RepressesROCKActivityandAltersActinDynamicsinMSCs.ROCKactivitydeclinedrapidlyduringBMP7-inducedbrownadipogenesisreachingundetectablelevelsinmatureadipocytes.ConsistentwithitsinhibitionofROCK,biochemicalandcellstainingassaysrevealedthatBMP7increasedtheG/F-actinratio.TheROCKinhibitorY27632similarlyenhancedG-actinlevelsrelativetoF-actin.第17页，课件共34页，创作于2023年2月ResultInhibitionoftheROCK/SRFPathwayEnhancesBrownAdipogenesisinMSCs.?Y27632couldmimictheeffectsofBMP71)Y27632aloneonlyhasamodesteffectonadipocytegeneexpressionandonnumberofcellsconvertingtoadipocytesthatcouldbeNileRedstained.2)Y27632didpotentiatetheactionofBMP7.(PPAR𝞬,FABP4↑)ROCKinhibitionpartiallyovercametheinhibitoryeffectofGremlinonadipocytegeneanddifferentiationwhilehavinglimitedeffectsonUCP1expression.✪Y27632couldnotexactly

mimicBMP7.★第18页，课件共34页，创作于2023年2月ResultInhibitionoftheROCK/SRFPathwayEnhancesBrownAdipogenesisinMSCs.➣CCG1423➞MSCs➞enhanceconversiontoadipocytes,andinduceexpressionofbrownadipocytesgenes.?

TheproadipogenicactivityofBMP7involvessuppressionofMRTF/SRFactivity★CCG1423didnotaltertheactincytoskeleton.➜CCG1423blocksSRFdownstreamofF-actinpolymerization.✪CCG1423couldmimicBMP7.★第19页，课件共34页，创作于2023年2月1BMP7andTGFbmediatedistincteffectsonlineagecommitmentofMSCsthroughalterationofROCKactivity.

。2BMP7RepressesROCKActivityandAltersActinDynamicsinMSCs.3MRTFAandSRFOverexpressionPreventDifferentiationofMSCstoAdipocytes.Results4IncreasedProductionofBeigeAdipocytesinWATofMRTFA-DeficientMice.5StromalVascularCellsfromMRTFA–/–iWATUndergoBeigeAdipogenesisMoreExtensivelyThanWTSVFs.

6MRTFA-DeficientMiceAreProtectedfromDiet-InducedObesityandInsulinResistance.

。7MRTFARegulatesWhole-BodyEnergyExpenditure.第20页，课件共34页，创作于2023年2月ResultMRTFAandSRFOverexpressionPreventDifferentiationofMSCstoAdipocytes.➣C3H/10T1/2MSCsdifferentiatetoadipocytes:TheexpressionofMRTFA,MRTFB,SRF,andSMA,collagen1a1,collagen3a1(SRFtargetgenes)↓TheexpressionofFABP4,UCP-1↑第21页，课件共34页，创作于2023年2月ResultMRTFAandSRFOverexpressionPreventDifferentiationofMSCstoAdipocytes.➣EctopicallyexpressSRForMRTFA:ThedifferentiationofMSCstoadipocytes↓Theexpressionofselectwhiteandbrownadipogenicgenes↓✪theROCK-SRFpathwayisanimportantBMP7target

✮ROCK-SRF↓

➜adipogenesis↑➣CCG1423couldpartiallyrescuedtheinhibitoryeffectofMRTFAorSRFonadipocyteformation.第22页，课件共34页，创作于2023年2月1BMP7andTGFbmediatedistincteffectsonlineagecommitmentofMSCsthroughalterationofROCKactivity.

。2BMP7RepressesROCKActivityandAltersActinDynamicsinMSCs.3MRTFAandSRFOverexpressionPreventDifferentiationofMSCstoAdipocytes.Results4IncreasedProductionofBeigeAdipocytesinWATofMRTFA-DeficientMice.5StromalVascularCellsfromMRTFA–/–iWATUndergoBeigeAdipogenesisMoreExtensivelyThanWTSVFs.

6MRTFA-DeficientMiceAreProtectedfromDiet-InducedObesityandInsulinResistance.

。7MRTFARegulatesWhole-BodyEnergyExpenditure.第23页，课件共34页，创作于2023年2月Result➣KOmouse●iWAT:moremultilocularadipocytes●eWAT:fewmultilocularadipocytes,smalleradipocytes.●BAT:nomorphologicalorUCP1expressionchanges.➣WTmouse●iWAT:hypertrophy.Biggerindividualunilocularadipocytes.➣Bodyweightdepotmorphology➣KOmouse●Lessweight&lessoverallfatmassrelativetoleanmass(6weeksoflowfatdiet).●Lowerfastinglevelsofglucose,butnodifferencesinglucose(aGTT).●Lowercirculatinglevelsofleptin&higherlevelsofadiponectinandinsulin.IncreasedProductionofBeigeAdipocytesinWATofMRTFA-DeficientMice.第24页，课件共34页，创作于2023年2月ResultIncreasedProductionofBeigeAdipocytesinWATofMRTFA-DeficientMice.►RNAanalysis➤iWATofKOmouse✓moreexpressionofUCP1mRNA✓moreexpressionofotherbrownadipocytemRNAs➤BATofKOmouse✓n.s.withWTmouseinUCP1mRNA,buthigherthaniWAT.

➤WATgenes➣AgtLowerlevelsofexpressioninKOmouseiWAT&higherdevelopmentofbrown-likeadipocytesinKOmouseiWAT➣SMALowerlevelsofexpressioninKOmouseiWAT➤Tbx1(amarkerofbeigeadipocyte)HigherlevelsofexpressioninKOmouseiWAT➽theappearanceofUCP1positive,multilocularadipocytesinKOmiceislikelyduetorecruitmentofbeigeprogenitors.第25页，课件共34页，创作于2023年2月1BMP7andTGFbmediatedistincteffectsonlineagecommitmentofMSCsthroughalterationofROCKactivity.

。2BMP7RepressesROCKActivityandAltersActinDynamicsinMSCs.3MRTFAandSRFOverexpressionPreventDifferentiationofMSCstoAdipocytes.Results4IncreasedProductionofBeigeAdipocytesinWATofMRTFA-DeficientMice.5StromalVascularCellsfromMRTFA–/–iWATUndergoBeigeAdipogenesisMoreExtensivelyThanWTSVFs.

6MRTFA-DeficientMiceAreProtectedfromDiet-InducedObesityandInsulinResistance.

。7MRTFARegulatesWhole-BodyEnergyExpenditure.第26页，课件共34页，创作于2023年2月ResultStromalVascularCellsfromMRTFA–/–iWATUndergoBeigeAdipogenesisMoreExtensivelyThanWTSVFCells.

?

MRTFArepressesbeigeadipocyteformationdirectly►SVFsofWTandKOiWAT➞adipocytes(invitro)

➣comparableexpressionofadiponectin,adipsin,FABP4.☞approximatelythesameextent.

➣KOSVFs:selectBATproteins(UCP1,Chchd10/NDG2)↑

➣FSK➞adipocytes➞expressionofUCP1mRNA(WT&KO)

➘UCP1,Fabp3,Cidea,Cox7a1,andElovl3⇈(KO)✤somebeigeadipocyteshaveasmoothmuscle-likeorigin.第27页，课件共34页，创作于2023年2月ResultStromalVascularCellsfromMRTFA–/–iWATUndergoBeigeAdipogenesisMoreExtensivelyThanWTSVFCells.

►BMP7➞SVFs➞MRTFA,CRP2,SMA,andcalponin↓★theKOSVFcells:loweramountofSMproteins

►TGF𝝱

➞SVFs➞SMprotein↑✪BMP7directsSVFprogenitorstoabeigeadipocytelineageratherthanaSMlineage.第28页，课件共34页，创作于2023年2月1BMP7andTGFbmediatedistincteffectsonlineagecommitmentofMSCsthroughalterationofROCKactivity.

。2BMP7RepressesROCKActivityandAltersActinDynamicsinMSCs.3MRTFAandSRFOverexpressionPreventDifferentiationofMSCstoAdipocytes.Results4IncreasedProductionofBeigeAdipocytesinWATofMRTFA-DeficientMice.5StromalVascularCellsfromMRTFA–/–iWATUndergoBeigeAdipogenesisMoreExtensivelyThanWTSVFs.

6MRTFA-DeficientMiceAreProtectedfromDiet-InducedObesityandInsulinResistance.

。7MRTFARegulatesWhole-BodyEnergyExpenditure.第29页，课件共34页，创作于2023年2月ResultMRTFA-DeficientMiceAreProtectedfromDiet-InducedObesityandInsulinResistance.►HFD:high-fatdietStartat6weeksagefor6weeks.➣KO🐭✓lessweight✓lessfatmass✓lessiWAT&eWATweight,nosignificantchangeinBAT✓lessliverweight☜mayduetotheextensiveaccumulationofhepaticlipidinWTmice.✓lowerlevelsoffastingglucose✓enhancedglucosetolerance✓lowercirculatinglevelsofleptin⇌lessfat&highercirculatinglevelsofadiponectin✓nosignofexcesslipiddepositionintheindividualadipocytesofiWATandeWAT✓iWATproducedabundantamountsofUCP1&otherbrowngenes✓areductionincrown-likestructuresintheeWAT☞KOmicewereprotectedfromdiet-inducedinflammation

✪MRTFAdeficiencyreducesHFD-inducedobesity,insulinresistance,eWATinfl