1、China-Japan Cardiovascular Forum 2019New Strategies for Immune-mediated Heart DiseasesMitsuakiIsobeDepartment of Cardiovascular MedicineTokyo Medical and Dental UniversityImmune-mediated heart diseasesIschemia-reperfusion injuryAtherosclerosisRestenosisCardiac allograft rejectionCardiac allograft va
2、sculopathyMyocarditisCongestive heart failureArrhythmiaImmune-mediated heart diseasesIschemia-reperfusion injuryAtherosclerosisRestenosisCardiac allograft rejectionCardiac allograft vasculopathyMyocarditisCongestive heart failureArrhythmiaInflammation in coronary arteriosclerosis and atherogenesisPP
3、AR(Peroxisome proliferator-activated receptor-)NFBAcute cardiac allograft rejectionEctopic heart transplantionrecipients:C3H/He donors:BALB/c full mismatchAdministration of pioglitazone(3mg/kg/day)to the recipient mice.Murine heart transplantationProlongation of heart graft by pioglitazone1Survival
4、Rate0.80.60.4*Pioglitazone(3mg/kg/day)Control chow0.2010203040506070*p0.05Postoperative DaysKosuge,Suzuki,Isobe:Circulation 113:2613-2622、2019Suppression of T cell Proliferation by Pioglitazone in Mixed Lymphocyte Reaction*0.8*Optical Density0.60.4*p0.050.20R:responder(recipient)S:stimulator(donor)P
5、io:pioglitazoneChronic cardiac rejectionGraft arterial disease(GAD)Accelerated coronary sclerosisGraft vasculopathyCardiac allograft vasculopathyMyocardiumRCASmallarteriesPathology:Diffuse intimalthickening of small to largecoronary arteriesProgression:months to yearsRisk factors:CMV infectionepisod
6、es of acute rejectionTherapy:?LAD8-y-o boyLCXChronic cardiac allograft rejectionCardiac allograft vasculopathyrecipients:C57BL/6 donors:Bm12 classII mismatch-10 7 14 21 28 35 42 49 56 daysControl chowPioglitazone(3mg/kg/day)Telmisartan(10mg/kg/day)sacrificeAttenuation of Graft Arteriosclerosis by Tr
7、eatment with Pioglitazone or Telmisartanp0.05ControlLuminal occlusion(%)TelmisartanControlPioglitazonep0.05Pioglitazone8 Weeks after TransplantationControlTelmisartanKosuge,Suzuki,Isobe:Circulation 2019SMC Proliferation AssaySMCsAortaPioglitazonetelmisartanactivated splenocytesProliferationSuppressi
8、on of SMC proliferation?Suppression of SMC Proliferation after Interaction with Splenocytes by Pioglitazone or Telmisartan*5.0*0.70.60.50.40.30.2*OpticalDensity4.03.02.01.000.10Summary 1?PPAR is associated with pathophysiology of immune-mediated heart diseases.?PPAR-agonists(pioglitazone and telmisa
9、rtan)are effective in suppressing experimental cardiac allograft rejection(acute rejection and allograft vasculopathy).ReferencesKosuge,Suzuki,Isobe:Circulation 113:2613-2622,2019Isobe,Kosuge,Suzuki:ATVB,26:1447-1456,2019Kosuge,Suzuki,Isobe:ATVB,26:2660-2665,2019Onai,Suzuki,Maejima:Am J Physiol,292,
10、H530-538,2019 Kosuge,Suzuki,Isobe,Transplanation,in pressMaejima,Suzuki,Okada,Isobe:in submissionInflammation in coronary arteriosclerosis and atherogenesisPPAR(Peroxisome proliferator-activated receptor-)NFBLPSTNFTCellTCRNFBRole of NF-kB in Cardiovascular DiseasesStimuliAnoxiaReactive oxygen specie
11、sLPSCytokines(IL-1,TNF)Re-oxygenationLipoproteinThrombinNF-B ActivationGene ExpressionCytokines(IL-1,TNF)iNOS,COX-2Adhesion molecules(ICAM-1,VCAM-1)Immunoreceptors(MHC class I,II)IL-6,M-CSF,GM-CSFChemokines(MCP-1,)Acute phase protein(CRP,)Heart failureVascular remodelingAllograft rejectionMyocarditi
12、sReperfusion injuryVentricular remodelingNF-B decoy-Mechanism of Action-StimuliStimuliI-?B kinasesI-?B kinasesInflammatoryInflammatorymoleculesmoleculesDegradationCell Cell MembraneMembraneI-Bp50p50p65p65PI-Bp50p50p65p65I-BXmRNANF-?Bp50p50p65p65p50p50p65p65DecoyDecoyNucleusNucleusp50p50p65p65XInflam
13、matory genesInflammatory genesCis-ElementCis-ElementAcute MyocarditisExperimental Allergic Myocarditis in RatsImmunizationLewis rats were immunized with porcine cardiac myosin.In vivo gene transferInjection of HVJ-liposome-NFB decoy at coronary cuspFITC-labeled Decoy in Rat HeartDay 0,7 or 1421 days
14、 after transfectionNFkB decoy reduces extent of autoimmune myocariditisControl(%)(%)10010080806060NFkB decoy404020200 0UTUTSDSDday0day0day7day7day14day14Myocarditis-affected area(injection at day 7)(Yokoseki O,Isobe M,Circ Res 2019)Method of gene transfer to cardiac allograftDonorHVJ-liposomeHVJ-Don
15、or HeartDecoy TransfectionTransfection:10 min,4CIschemic time:30 minRecipient28 daysHistological analysisEffect of NFB decoy on GADEvG stainingPDGF-B mRNAVCAM-1ScrambledecoyNFB decoy(Suzuki,Isobe,Gene Therapy,2019)Gene Transfer SystemDecoy+microbubble complexUltrasound(US)Wire injuryInhibition of in
16、timal hyperplasia by NF-kB decoy transfectionNeointimal formation(I/M)I/M2p 75%by QCA)NegativePositivePatients171Before PCIAfter PCIRates94.1%5.9%6 Mo.LaterAverage%Restenosis（by QCA）%Restenosis88.51.539.6ConclusionsPPAR and NFB are good molecular targets for treatment of immune-mediated cardiovascul
17、ar diseases.Further studies should be conducted to explore the possibility to develop new therapeutic strategies.AcknowledgementTokyo Medical and Dental UniversityJun-ichi SuzukiHisanori Kosuge Noritaka KogaNoriko TamuraMasahito Ogawa Ryuichi MorishitaYasufumi KanedaKensuke EgashiraHiroshi ItoOsaka
18、UniversityKyushu UniversityAkita UniversitySuppression of Cytokines Expression in Allografts by Treatment with PioglitazoneRNase protection assay IL-10IL-15IL-6IFN-5 days after transplantationSuppression of T cell Proliferation by Telmisartan in Mixed Lymphocyte Reaction0.8*Optical Density0.60.40.20
19、*p0.05R:responder(recipient)S:stimulator(donor)Tel:telmisartanUltrasound+microbubbles increase efficacy of transfectionFITC Positive Cells per Arteryp0.05No Ultrasound*200100Decoy(+)Microbubble(-)Ultrasound(+)(+)(+)(-)(+)(+)(+)Ultrasound(Suzuki J,Isobe M,J Vasc Res,2019)Suppression of Neointimal Formation by NF-B Decoy(%)100*P P 0.01 0.01*Minor mismatch(DBA to B10D2)500NF-kBDecoyE2FDecoyAntisenseNo ODNcdk2 ODN28 DaysVascular remodeling after injuryTransfection without viral vectorMicrobubbles with ultrasound
