Partial Reprogramming of Pluripotent Stem Cell-Derived Cardiomyocytes into Neurons.
Direct reprogramming of somatic cells has been demonstrated, however, it is unknown whether electrophysiologically-active somatic cells derived from separate germ layers can be interconverted. We demonstrate that partial direct reprogramming of mesoderm-derived cardiomyocytes into neurons is feasible, generating cells exhibiting structural and electrophysiological properties of both cardiomyocytes and neurons. Human and mouse pluripotent stem cell-derived CMs (PSC-CMs) were transduced with the neurogenic transcription factors Brn2, Ascl1, Myt1l and NeuroD. We found that CMs adopted neuronal morphologies as early as day 3 post-transduction while still retaining a CM gene expression profile. At week 1 post-transduction, we found that reprogrammed CMs expressed neuronal markers such as Tuj1, Map2, and NCAM. At week 3 post-transduction, mature neuronal markers such as vGlut and synapsin were observed. With single-cell qPCR, we temporally examined CM gene expression and observed increased expression of neuronal markers Dcx, Map2, and Tubb3. Patch-clamp analysis confirmed the neuron-like electrophysiological profile of reprogrammed CMs. This study demonstrates that PSC-CMs are amenable to partial neuronal conversion, yielding a population of cells exhibiting features of both neurons and CMs.
Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA.|Cardiovascular Center, Far Eastern Memorial Hospital, New Taipei City, Taiwan.
Wenpo
W
Chuang
Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA.|Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA.
Arun
A
Sharma
Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA.|Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA.
Praveen
P
Shukla
Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA.|Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA.
Guang
G
Li
Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA.
Moritz
M
Mall
Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA.|Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA.
Kuppusamy
K
Rajarajan
Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA.|Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, CA, USA.
Oscar J
OJ
Abilez
Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA.|Department of Biology, Stanford University School of Medicine, Stanford, CA, USA.
Ryoko
R
Hamaguchi
Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA.|Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA.|Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, CA, USA.|Department of Radiology, Molecular Imaging Program, Stanford University School of Medicine, Stanford, CA, USA.
Joseph C
JC
Wu
Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA.|Department of Pathology, Stanford University School of Medicine, Stanford, CA, USA.
Marius
M
Wernig
Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA.|Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, USA.|Department of Medicine, Division of Cardiology, Stanford University School of Medicine, Stanford, CA, USA.
Sean M
SM
Wu
D000818:Animals; D015415:Biomarkers; D002454:Cell Differentiation; D002460:Cell Line; D065150:Cellular Reprogramming; D000089662:Doublecortin Protein; D055724:Electrophysiological Phenomena; D053595:Embryonic Stem Cells; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D017930:Genes, Reporter; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism*; D009474:Neurons / Q000166:cytology* / Q000378:metabolism*; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D014161:Transduction, Genetic
D015415:Biomarkers; C000714847:Dcx protein, mouse; D000089662:Doublecortin Protein