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1、In Vivo Cellular and Molecular MRI of Embryonic Stem Cell Survival and ProliferationPhillip C. Yang, MD Assistant Professor Division of Cardiovascular Medicine Stanford University (No disclosure or conflict of interest)GREAT WALL INTERNATIONAL CONGRESS OF CARDIOLOGY BEIJING, CHINA 2008I. Stem Cell T
2、herapy: Congestive Heart Failure Coronary artery disease: #1 killer in the United States 1.1 million new or recurrent MI and 500,000 deaths per year Improved acute MI therapy shift in the landscape Congestive heart failure: #1 cause of hospital admissions in the US Prevalence: 5 million patients Inc
3、idence: 300,000 patients/year Advanced therapy: 5-year survival 50% Heart transplant: 1,000 pt/year Recent stem cell research: alternative therapeutic option?II. Evaluation of Cell TherapyClinical trials: modest, temporary improvement in LVEF Cell survival following transplantation into the heart Fu
4、ndamental biology of cell therapy: Restore & regenerate In vivo imaging of stem cell viabilityIII. In Vivo Multi-Modality ImagingOptical Imagingd2d3d4d5d6Optical Imagingd2d3d4d5d6Optical Imagingd2d3d4d5d6Optical Imagingd2d3d4d5d6Optical Imagingd2d3d4d5d6MRIRNBLI Sensitivity 10-4 - 10-9 Mole/L10-11 -
5、10-12 Mole/L10-15 - 10-17 Mole/LSpatial resolution500 m3-5 mm3-5mmTemporal resolution10 msseconds seconds(Hendry et al, JCTS, 2008)In vivo molecular and cellular MRICellCellNucleusVector (Delivery Vehicle)ProteinReporter GeneIron-oxide labeld MAbIRESCMV/ubiquitinIgKHA/mycPDGFR-TMGFPlucmRNAPLL PSPSPS
6、 PLLPSPLLPLLIron-oxide HAmycManganeseCa2+-channel IV. Cellular MRI: iron-oxideIV. Cellular MRI: iron-oxideH&EPrus blueH&E 40xP blue 40x(Hung et al, Circulation: Cardiovascular Img. 2008)(Suzuki et al, MRM, 2007)Metabolic effects: 3 transfection methodsIn vivo MRI of myocardial restoration End- diast
7、oleEnd- systoleMI controlNon-treated heartTreated heart (Hendry et al, JCTS 2008)mESC mESC FibroblastCellular survival(Cunningham et al, Magn Reson Med 2005)Survivald2d5d15d2d5d7GREORIn vivo serial survival and proliferation1POD4POD7POD10POD16POD456 (Suzuki et al, MRM, accepted, 2008)Control0.010.05
8、0.100.501.00(mM)p/sec/cm2/srV. Cellular MRI: Manganese3.000.01 with Dead cells(Yamada et al, MRM, submitted)Biological modulation of Ca2+-channel activitycontrolMnMn + BKMn+Vera727379122097.2147.6Non-contractile embryoid bodyContractile embryoid bodyIn vivo Mn2+ MRI of myocardial survival and terato
9、maVI. In vivo molecular imaging: Reporter GeneCellCellNucleusDNA Reporter Gene ConstructPromoterReporter GeneVector (Delivery Vehicle)mRNAReporter Protein ProbeDetectorSignalIn vivo molecular MRI: SurvivalUSPIO-monoclonal Ab against HA & c-mycMRI reporter gene driven by CMV/ubiquitin promoterCloning
10、 into p2k7-RG lentivirusUbc EF1p2k7(neo or blas)Ubc EF1(Chung et al, Nat Biotech, 2008, in prep)Validation: FACS c-myc expressionHA expressionmESChESC Validation: ImmunohistologyAnti c-myc FITCDAPIPrussian blueValidation: In vitro BLI and MRImESCmESC + anti c-myc mESC-RG + anti c-mycmESC-RG + anti H
11、AmESC + anti HAmESC-RGhESC + anti c-mychESC-RG + anti c-mychESC + anti HAhESC-RG + anti HAIn vivo MRI of hESC-RGVII. ConclusionIn vivo cellular and molecular MRI: iron-oxide, manganese, and reporter geneRobust labeling Safety profile Dual imaging High sensitivity: longitudinal tracking of location,
12、survival, and teratoma High spatial and temporal resolution: myocardial restorationStanford Laboratory for Cardiovascular RegenerationYang LaboratoryTakayasu Arai Jaehoo Chung Rajesh Dash Pratima Kundu Yoriyasu Suzuki Mayumi Yamada Hunter WangCT Surgery Stephen Hendry Grant Hoyt Ahmed Sheikh Robert
13、RobbinsCV Molecular Biology Thomas QuertermousCV Imaging Feng Cao Chuck Cunningham Tim Doyle Sam Gambhir Michael McConnell John Pauly Masahiro Terashima Joe Wu Esther XiePathology Andrew ConnollyCV Intervention Todd Brinton Peter Fitzgerald Alan Yeung Paul YockStem Cell Biology Renee Reijo-Pera Micha Drukker Irv WeissmanFunding Source NIH AHA CIRM
