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Series GSE54022 Query DataSets for GSE54022
Status Public on Jan 14, 2014
Title Inhibition of TGFβ Signaling Increases Direct Conversion of Fibroblasts to Induced Cardiomyocytes
Organism Mus musculus
Experiment type Expression profiling by array
Summary Recent studies have been successful at utilizing ectopic expression of transcription factors to generate induced cardiomyocytes (iCMs) from fibroblasts, albeit at a low frequency in vitro. This work investigates the influence of small molecules that have been previously reported to improve differentiation to cardiomyocytes as well as reprogramming to iPSCs in conjunction with ectopic expression of the transcription factors Hand2, Nkx2.5, Gata4, Mef2C, and Tbx5 on the conversion to functional iCMs. We utilized a reporter system in which the calcium indicator GCaMP is driven by the cardiac Troponin T promoter to quantify iCM yield. The TGFβ inhibitor, SB431542 (SB), was identified as a small molecule capable of increasing the conversion of both mouse embryonic fibroblasts and adult cardiac fibroblasts to iCMs up to ~5 fold. Further characterization revealed that inhibition of TGFβ by SB early in the reprogramming process led to the greatest increase in conversion of fibroblasts to iCMs in a dose-responsive manner. Global transcriptional analysis at Day 3 post-induction of the transcription factors revealed an increased expression of genes associated with the development of cardiac muscle in the presence of SB compared to the vehicle control. Incorporation of SB in the reprogramming process increases the efficiency of iCM generation, one of the major goals necessary to enable the use of iCMs for discovery-based applications and for the clinic.
 
Overall design Mouse embryonic fibroblasts (MEFs) and adult mouse cardiac fibroblasts (CFs) were transfected with an empty vector (0F) or the combination of Hand2, Nkx2.5, Gata4, Mef2C, and Tbx5 (5F). Samples were exposed to the vehicle control (D, DMSO), SB431542 (SB, 0.5 uM MEF, 5 uM CF), or TGFb1 (T, 2 ng/mL) during culture. Transcription factor expression was induced at Day 0 and samples were isolated at Day 3 post-induction.
 
Contributor(s) Ifkovits JL, Addis RC, Epstein JA, Gearhart JD
Citation(s) 24586958
Submission date Jan 13, 2014
Last update date Mar 04, 2019
Contact name John Gearhart
E-mail(s) gearhart@upenn.edu
Phone 215-898-4450
Organization name University of Pennsylvania
Department Dept of Cell and Developmental Biology, Institute for Regenerative Medicine
Street address 3400 Civic Center Blvd, Building 421, 9-121 Smilow Center for Translational Research
City Philadelphia
State/province PA
ZIP/Postal code 19104
Country USA
 
Platforms (1)
GPL6246 [MoGene-1_0-st] Affymetrix Mouse Gene 1.0 ST Array [transcript (gene) version]
Samples (27)
GSM1305950 MEF 0F DMSO Rep 1
GSM1305951 MEF 0F DMSO Rep 2
GSM1305952 MEF 0F DMSO Rep 3
Relations
BioProject PRJNA234369

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE54022_RAW.tar 130.8 Mb (http)(custom) TAR (of CEL, CHP)
Raw data provided as supplementary file
Processed data included within Sample table
Processed data provided as supplementary file

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