be230cLecture5

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Unformatted text preview: Cardiomyocyte DifferenJaJon Assays to Discover and Characterize Small Molecules for RegeneraJve ApplicaJons Screening IdenJfies Small Molecules that Promote DifferenJaJon of Cardiomyocytes From MulJpotent hESC Progenitors DAPI, eGFP Flk1+, MesP1+ MulJpotent Cardiogenic Progenitor Cardiomyocyte Mesendoderm Progenitor Vascular endothelial cells Neg. control Vascular smooth muscle Compounds Day 4 Day 9 Pos. control Flow Cytometry: AcJve Series Example Control (DMSO vehicle) MLG ­III ­025AH cardiomyocytes Day 10 Willems et al., CirculaJon Res. 2011; Willems et al., Cell Stem Cell, 2012 13  80 ITD 1(+) ITD 1 0.01 0.1 1 4/15/13 10 ITD 1 ITD 1( )/ITD 1(+) ncentration(µM) ITD 2 ITDts L Lefty1expressionlevel          ITDts ITD 2 ITD 1 SB  ITD 1(+)  1400 140 1200 120 Candidate Molecules 1000 100 •  Cardiac regeneraJon (Wnt inhibitor, 800 Willems et al., Circ Res 2011) •  Cardiac regeneraJon (first selecJve 600 TGFβ inhibitor; Willems et al., Cell Stem Cell 2012) 400 •  Highly potent TCF inhibitor is not 200 very acJve in cardiogenesis, but 0 under evaluaJon for prostate cancer ActivinA •  Inhibitor of TriC chaperone protein re ­folding ITD 15µM acJve in cardiogenesis •  AcJvator of cSB5µM ardiomyocyte Ca2+ handling and contracJlity ActivinA pound 80 60 40 20 0 TGF 2 ITD 15µM SB5µM Bruce Conklin, UCSF TGF 2  IC50( M) Emax(%Inh) IC50( M) Emax(%Inh) 1.51±0.52 2.52±0.94 1.00±0.32 2.60±2.2 3.50±3.10 0.17±0.02 65±2 51±5 57±5 53±4 21±10 99±1 0.85±0.30 6.90±0.34 0.46±...
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This document was uploaded on 04/09/2014.

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