Advancing Regenerative Medicine with The Boston University ......Wellington Cardoso, MD, PhD Joyce...
Transcript of Advancing Regenerative Medicine with The Boston University ......Wellington Cardoso, MD, PhD Joyce...
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www.bumc.bu.edu/stemcellswww.bumc.bu.edu/stemcells
Advancing Regenerative Medicine withThe Boston University�Induced Pluripotent Stem (iPS) Cell Bank
Darrell N. Kotton, MDGustavo Mostoslavsky, MD, PhD
George Murphy, PhDWellington Cardoso, MD, PhD
Joyce Wong, PhDFeb. 16, 2010
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iPS Cell Bank
• What is it?
• Why is it important?
• How far off are we from using the Bank for Regenerative Medicine applications?
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Embryonic Stem Cells Embryonic Stem Cells -- ESCESC
BlastocystBlastocyst
ES cell colonyES cell colony
55--6 d6 daysays
Fertilized eggFertilized egg
EndodermEndodermEndoderm
MesodermMesodermMesoderm
EctodermEctodermEctoderm
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ESC Major Limitations for Regenerative MedicineESC Major Limitations for Regenerative Medicine
ESCESC iPS cellsiPS cells
Technical hurdlesTechnical hurdles
Ethical concernsEthical concerns
ImmunogenicityImmunogenicity
ESCESC
Technical hurdlesTechnical hurdles
Ethical concernsEthical concerns
ImmunogenicityImmunogenicity
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‘Reprogramming’
ESCell
PluripotentDifferentiatedSkin Fibroblasts
iPSCell
Takahashi and Yamanaka.Cell 2006.
The Discovery of Induced Pluripotent Stem (iPS) Cells
Oct4Sox2Klf4
cMyc
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‘Reprogramming’ with STEMCCA
55’’ LTRLTR PSIPSI RRERREHIV cpPuHIV cpPu
Oct4Oct4 F2AF2A Klf4Klf4 IRESIRES Sox2Sox2 E2AE2A cMyccMyc
WPREWPREdU3dU3
33’’ LTRLTR
STEMCCASTEMCCA
EF1EF1 or TetOor TetO
Tail Fibroblast
iPSCell
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iPS derivation without the presence of exogenous transgenesiPS derivation without the presence of exogenous transgenes
55’’ LTRLTR PSIPSI RRERREHIV cpPuHIV cpPu
Oct4Oct4 F2AF2A Klf4Klf4 IRESIRES Sox2Sox2 E2AE2A cMyccMyc
WPREWPREdU3dU3
33’’ LTRLTR
STEMCCASTEMCCA
EF1EF1 or TetOor TetO
loxPloxP
STEMCCASTEMCCA
EF1EF1
loxploxp
55’’ LTRLTR PSIPSI RRERREHIV cpPuHIV cpPu
Oct4Oct4 F2AF2A Klf4Klf4 IRESIRES Sox2Sox2 E2AE2A cMyccMycWPREWPRE
dU3dU333’’ LTRLTR
loxploxpSTEMCCASTEMCCA
55’’ LTRLTR PSIPSI RRERREHIV cpPuHIV cpPu
EF1EF1
Oct4Oct4 F2AF2A Klf4Klf4 IRESIRES Sox2Sox2 E2AE2A
WPREWPREdU3dU3
33’’ LTRLTR
loxploxp
dU3dU3
ChromChrom ChromChrom
dU3dU3
ChromChrom
loxploxpChromChrom
Reverse Transcription Reverse Transcription + integration + integration
Cre excisionCre excision
cMyccMyc
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Boston University iPS Cell Bank
• What is it?- A repository of human iPS cells generated
from individuals affected by diseases treated or studied at BUSM/BMC
•Why is it important?
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The Potential of iPS Research The Potential of iPS Research
•• Skin fibroblastsSkin fibroblasts•• Blood cellsBlood cells•• Hair keratinocytesHair keratinocytes•• ????
iPS cellsiPS cells
cell therapycell therapy
Normal embryo developmentNormal embryo developmentDisease modelsDisease modelsDrug screeningDrug screening
Differentiation into
Differentiation into
specific lineages
specific lineages
Emphysema (AAT Def.)Cystic Fibrosis Diabetes
IBD Sickle Cell AmyloidosisLong QT Syndrome
Transduction/Transduction/TransfectionTransfection
Nucle
ar Re
progra
mming
Nucle
ar Re
progra
mming Oct4
Klf4 Sox2
cMyc
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Boston University iPS Cell Bank
• What is it?- A repository of human iPS cells generated
from individuals affected by diseases treated or studied at BUSM/BMC
•Why is it important?-Model disease/development-Develop cell-based therapies
•How far off are we from using the Bank for Regenerative Medicine applications?
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ARC StructureSkin biopsyFibroblasts
Reprogram
STEMCCA
iPS Cell Bank
EndodermLungLiverGutPancreas
Kotton, Mostoslavsky,Cardoso
MesodermBloodEndotheliumCardiomyocytesOther mesenchyme
Murphy, MostoslavskyWong, Keller
In vivo DeliveryScaffoldsBiocapsules
BU BMEWong
Tra1-81
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Cystic Fibrosis
JC JeanAba SomersChris FordAmel Omari
Emphysema(AAT Deficiency)
Scleroderma
Long QT Syndrome
Disease-Specific Human iPS Cell LinesAmyloid (TTR)
Sickle Cell Dz
Brenden SmithGeorge MurphyCesar SommerBob Lafyatis
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Regenerative Medicine ARCDarrell Kotton Lab
George Murphy Lab
www.bumc.bu.edu/stemcellswww.bumc.bu.edu/stemcells
Joyce Wong LabWellington Cardoso Lab
Gustavo Mostoslavsky Lab