TY - JOUR
T1 - Distinct roles of microRNA-1 and -499 in ventricular specification and functional maturation of human embryonic stem Cell-Derived cardiomyocytes
AU - Fu, Ji Dong
AU - Rushing, Stephanie N.
AU - Lieu, Deborah K.
AU - Chan, Camie W.
AU - Kong, Chi Wing
AU - Geng, Lin
AU - Wilson, Kitchener D.
AU - Chiamvimonvat, Nipavan
AU - Boheler, Kenneth R.
AU - Wu, Joseph C.
AU - Keller, Gordon
AU - Hajjar, Roger J.
AU - Li, Ronald A.
N1 - Funding Information:
This work has received support from VistaGen Therapeutics, Inc., South San Francisco, CA. There are no patents, products in development or marketed products to declare. This does not alter the authors' adherence to all the PLoS ONE policies on sharing data and materials, as detailed online in the guide for authors.
PY - 2011/11/16
Y1 - 2011/11/16
N2 - Background: MicroRNAs (miRs) negatively regulate transcription and are important determinants of normal heart development and heart failure pathogenesis. Despite the significant knowledge gained in mouse studies, their functional roles in human (h) heart remain elusive. Methods and Results: We hypothesized that miRs that figure prominently in cardiac differentiation are differentially expressed in differentiating, developing, and terminally mature human cardiomyocytes (CMs). As a first step, we mapped the miR profiles of human (h) embryonic stem cells (ESCs), hESC-derived (hE), fetal (hF) and adult (hA) ventricular (V) CMs. 63 miRs were differentially expressed between hESCs and hE-VCMs. Of these, 29, including the miR-302 and -371/372/373 clusters, were associated with pluripotency and uniquely expressed in hESCs. Of the remaining miRs differentially expressed in hE-VCMs, 23 continued to express highly in hF- and hA-VCMs, with miR-1, -133, and -499 displaying the largest fold differences; others such as miR-let-7a, -let-7b, -26b, -125a and -143 were non-cardiac specific. Functionally, LV-miR-499 transduction of hESC-derived cardiovascular progenitors significantly increased the yield of hE-VCMs (to 72% from 48% of control; p<0.05) and contractile protein expression without affecting their electrophysiological properties (p>0.05). By contrast, LV-miR-1 transduction did not bias the yield (p>0.05) but decreased APD and hyperpolarized RMP/MDP in hE-VCMs due to increased I to, I Ks and I Kr, and decreased I f (p<0.05) as signs of functional maturation. Also, LV-miR-1 but not -499 augmented the immature Ca 2+ transient amplitude and kinetics. Molecular pathway analyses were performed for further insights. Conclusion: We conclude that miR-1 and -499 play differential roles in cardiac differentiation of hESCs in a context-dependent fashion. While miR-499 promotes ventricular specification of hESCs, miR-1 serves to facilitate electrophysiological maturation.
AB - Background: MicroRNAs (miRs) negatively regulate transcription and are important determinants of normal heart development and heart failure pathogenesis. Despite the significant knowledge gained in mouse studies, their functional roles in human (h) heart remain elusive. Methods and Results: We hypothesized that miRs that figure prominently in cardiac differentiation are differentially expressed in differentiating, developing, and terminally mature human cardiomyocytes (CMs). As a first step, we mapped the miR profiles of human (h) embryonic stem cells (ESCs), hESC-derived (hE), fetal (hF) and adult (hA) ventricular (V) CMs. 63 miRs were differentially expressed between hESCs and hE-VCMs. Of these, 29, including the miR-302 and -371/372/373 clusters, were associated with pluripotency and uniquely expressed in hESCs. Of the remaining miRs differentially expressed in hE-VCMs, 23 continued to express highly in hF- and hA-VCMs, with miR-1, -133, and -499 displaying the largest fold differences; others such as miR-let-7a, -let-7b, -26b, -125a and -143 were non-cardiac specific. Functionally, LV-miR-499 transduction of hESC-derived cardiovascular progenitors significantly increased the yield of hE-VCMs (to 72% from 48% of control; p<0.05) and contractile protein expression without affecting their electrophysiological properties (p>0.05). By contrast, LV-miR-1 transduction did not bias the yield (p>0.05) but decreased APD and hyperpolarized RMP/MDP in hE-VCMs due to increased I to, I Ks and I Kr, and decreased I f (p<0.05) as signs of functional maturation. Also, LV-miR-1 but not -499 augmented the immature Ca 2+ transient amplitude and kinetics. Molecular pathway analyses were performed for further insights. Conclusion: We conclude that miR-1 and -499 play differential roles in cardiac differentiation of hESCs in a context-dependent fashion. While miR-499 promotes ventricular specification of hESCs, miR-1 serves to facilitate electrophysiological maturation.
UR - http://www.scopus.com/inward/record.url?scp=81155134381&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=81155134381&partnerID=8YFLogxK
U2 - 10.1371/journal.pone.0027417
DO - 10.1371/journal.pone.0027417
M3 - Article
C2 - 22110643
AN - SCOPUS:81155134381
SN - 1932-6203
VL - 6
JO - PloS one
JF - PloS one
IS - 11
M1 - e27417
ER -