title: Proteomic Analysis of Human Pluripotent Stem Cell-Derived, Fetal, and Adult Ventricular Cardiomyocytes Reveals Pathways Crucial for Cardiac Metabolism and Maturation.
abstract: BACKGROUND
  Differentiation of pluripotent human embryonic stem cells (hESCs) to the cardiac lineage represents a potentially unlimited source of ventricular cardiomyocytes (VCMs), but hESC-VCMs are developmentally immature. Previous attempts to profile hESC-VCMs primarily relied on transcriptomic approaches, but the global proteome has not been examined. Furthermore, most hESC-CM studies focus on pathways important for cardiac differentiation, rather than regulatory mechanisms for CM maturation. We hypothesized that gene products and pathways crucial for maturation can be identified by comparing the proteomes of hESCs, hESC-derived VCMs, human fetal and human adult ventricular and atrial CMs.
  
  
  RESULTS
  Using two-dimensional-differential-in-gel electrophoresis, 121 differentially expressed (>1.5-fold; P<0.05) proteins were detected. The data set implicated a role of the peroxisome proliferator-activated receptor α signaling in cardiac maturation. Consistently, WY-14643, a peroxisome proliferator-activated receptor α agonist, increased fatty oxidative enzyme level, hyperpolarized mitochondrial membrane potential and induced a more organized morphology. Along this line, treatment with the thyroid hormone triiodothyronine increased the dynamic tension developed in engineered human ventricular cardiac microtissue by 3-fold, signifying their maturation.
  
  
  CONCLUSIONS
  We conclude that the peroxisome proliferator-activated receptor α and thyroid hormone pathways modulate the metabolism and maturation of hESC-VCMs and their engineered tissue constructs. These results may lead to mechanism-based methods for deriving mature chamber-specific CMs.
authors.affiliation: From the Stem Cell and Regenerative Medicine Consortium (E.P., W.K., B.Y., Z.W., O.T. W., K.R.B., R.A.L.) and Department of Physiology, LKS Faculty of Medicine (E.P., W.K., B.Y., Z.W., O.T.W., K.R.B., R.A.L.), University of Hong Kong, Hong Kong, P.R. China; Departments of Biology and Chemistry (Y.M.L., R.R., Y.W.L.) and Computer Science (H.S.W.), City University of Hong Kong, Hong Kong, P.R. China; Department of Computer Science, Guangzhou University, Guangzhou, P.R. China (S.Z.); Department of Bioengineering, Boston University, MA (A.C., C.C.); Harvard Wyss Institute for Biologically Inspired Engineering, Boston, MA (A.C., C.C.); Department of Cell Biology and Human Anatomy, University of California, Davis (J.M., A.H., D.K.L.); Cardiovascular Research Center, Mount Sinai School of Medicine, New York (D.K.L., R.A.L.); and Division of Cardiology, Johns Hopkins University, Baltimore, MD (G.F.T., K.R.B.).
authors.forename: Ellen
authors.initials: E
authors.lastname: Poon
authors.affiliation: From the Stem Cell and Regenerative Medicine Consortium (E.P., W.K., B.Y., Z.W., O.T. W., K.R.B., R.A.L.) and Department of Physiology, LKS Faculty of Medicine (E.P., W.K., B.Y., Z.W., O.T.W., K.R.B., R.A.L.), University of Hong Kong, Hong Kong, P.R. China; Departments of Biology and Chemistry (Y.M.L., R.R., Y.W.L.) and Computer Science (H.S.W.), City University of Hong Kong, Hong Kong, P.R. China; Department of Computer Science, Guangzhou University, Guangzhou, P.R. China (S.Z.); Department of Bioengineering, Boston University, MA (A.C., C.C.); Harvard Wyss Institute for Biologically Inspired Engineering, Boston, MA (A.C., C.C.); Department of Cell Biology and Human Anatomy, University of California, Davis (J.M., A.H., D.K.L.); Cardiovascular Research Center, Mount Sinai School of Medicine, New York (D.K.L., R.A.L.); and Division of Cardiology, Johns Hopkins University, Baltimore, MD (G.F.T., K.R.B.).
authors.forename: Wendy
authors.initials: W
authors.lastname: Keung
authors.affiliation: From the Stem Cell and Regenerative Medicine Consortium (E.P., W.K., B.Y., Z.W., O.T. W., K.R.B., R.A.L.) and Department of Physiology, LKS Faculty of Medicine (E.P., W.K., B.Y., Z.W., O.T.W., K.R.B., R.A.L.), University of Hong Kong, Hong Kong, P.R. China; Departments of Biology and Chemistry (Y.M.L., R.R., Y.W.L.) and Computer Science (H.S.W.), City University of Hong Kong, Hong Kong, P.R. China; Department of Computer Science, Guangzhou University, Guangzhou, P.R. China (S.Z.); Department of Bioengineering, Boston University, MA (A.C., C.C.); Harvard Wyss Institute for Biologically Inspired Engineering, Boston, MA (A.C., C.C.); Department of Cell Biology and Human Anatomy, University of California, Davis (J.M., A.H., D.K.L.); Cardiovascular Research Center, Mount Sinai School of Medicine, New York (D.K.L., R.A.L.); and Division of Cardiology, Johns Hopkins University, Baltimore, MD (G.F.T., K.R.B.).
authors.forename: Yimin
authors.initials: Y
authors.lastname: Liang
authors.affiliation: From the Stem Cell and Regenerative Medicine Consortium (E.P., W.K., B.Y., Z.W., O.T. W., K.R.B., R.A.L.) and Department of Physiology, LKS Faculty of Medicine (E.P., W.K., B.Y., Z.W., O.T.W., K.R.B., R.A.L.), University of Hong Kong, Hong Kong, P.R. China; Departments of Biology and Chemistry (Y.M.L., R.R., Y.W.L.) and Computer Science (H.S.W.), City University of Hong Kong, Hong Kong, P.R. China; Department of Computer Science, Guangzhou University, Guangzhou, P.R. China (S.Z.); Department of Bioengineering, Boston University, MA (A.C., C.C.); Harvard Wyss Institute for Biologically Inspired Engineering, Boston, MA (A.C., C.C.); Department of Cell Biology and Human Anatomy, University of California, Davis (J.M., A.H., D.K.L.); Cardiovascular Research Center, Mount Sinai School of Medicine, New York (D.K.L., R.A.L.); and Division of Cardiology, Johns Hopkins University, Baltimore, MD (G.F.T., K.R.B.).
authors.forename: Rajkumar
authors.initials: R
authors.lastname: Ramalingam
authors.affiliation: From the Stem Cell and Regenerative Medicine Consortium (E.P., W.K., B.Y., Z.W., O.T. W., K.R.B., R.A.L.) and Department of Physiology, LKS Faculty of Medicine (E.P., W.K., B.Y., Z.W., O.T.W., K.R.B., R.A.L.), University of Hong Kong, Hong Kong, P.R. China; Departments of Biology and Chemistry (Y.M.L., R.R., Y.W.L.) and Computer Science (H.S.W.), City University of Hong Kong, Hong Kong, P.R. China; Department of Computer Science, Guangzhou University, Guangzhou, P.R. China (S.Z.); Department of Bioengineering, Boston University, MA (A.C., C.C.); Harvard Wyss Institute for Biologically Inspired Engineering, Boston, MA (A.C., C.C.); Department of Cell Biology and Human Anatomy, University of California, Davis (J.M., A.H., D.K.L.); Cardiovascular Research Center, Mount Sinai School of Medicine, New York (D.K.L., R.A.L.); and Division of Cardiology, Johns Hopkins University, Baltimore, MD (G.F.T., K.R.B.).
authors.forename: Bin
authors.initials: B
authors.lastname: Yan
authors.affiliation: From the Stem Cell and Regenerative Medicine Consortium (E.P., W.K., B.Y., Z.W., O.T. W., K.R.B., R.A.L.) and Department of Physiology, LKS Faculty of Medicine (E.P., W.K., B.Y., Z.W., O.T.W., K.R.B., R.A.L.), University of Hong Kong, Hong Kong, P.R. China; Departments of Biology and Chemistry (Y.M.L., R.R., Y.W.L.) and Computer Science (H.S.W.), City University of Hong Kong, Hong Kong, P.R. China; Department of Computer Science, Guangzhou University, Guangzhou, P.R. China (S.Z.); Department of Bioengineering, Boston University, MA (A.C., C.C.); Harvard Wyss Institute for Biologically Inspired Engineering, Boston, MA (A.C., C.C.); Department of Cell Biology and Human Anatomy, University of California, Davis (J.M., A.H., D.K.L.); Cardiovascular Research Center, Mount Sinai School of Medicine, New York (D.K.L., R.A.L.); and Division of Cardiology, Johns Hopkins University, Baltimore, MD (G.F.T., K.R.B.).
authors.forename: Shaohong
authors.initials: S
authors.lastname: Zhang
authors.affiliation: From the Stem Cell and Regenerative Medicine Consortium (E.P., W.K., B.Y., Z.W., O.T. W., K.R.B., R.A.L.) and Department of Physiology, LKS Faculty of Medicine (E.P., W.K., B.Y., Z.W., O.T.W., K.R.B., R.A.L.), University of Hong Kong, Hong Kong, P.R. China; Departments of Biology and Chemistry (Y.M.L., R.R., Y.W.L.) and Computer Science (H.S.W.), City University of Hong Kong, Hong Kong, P.R. China; Department of Computer Science, Guangzhou University, Guangzhou, P.R. China (S.Z.); Department of Bioengineering, Boston University, MA (A.C., C.C.); Harvard Wyss Institute for Biologically Inspired Engineering, Boston, MA (A.C., C.C.); Department of Cell Biology and Human Anatomy, University of California, Davis (J.M., A.H., D.K.L.); Cardiovascular Research Center, Mount Sinai School of Medicine, New York (D.K.L., R.A.L.); and Division of Cardiology, Johns Hopkins University, Baltimore, MD (G.F.T., K.R.B.).
authors.forename: Anant
authors.initials: A
authors.lastname: Chopra
authors.affiliation: From the Stem Cell and Regenerative Medicine Consortium (E.P., W.K., B.Y., Z.W., O.T. W., K.R.B., R.A.L.) and Department of Physiology, LKS Faculty of Medicine (E.P., W.K., B.Y., Z.W., O.T.W., K.R.B., R.A.L.), University of Hong Kong, Hong Kong, P.R. China; Departments of Biology and Chemistry (Y.M.L., R.R., Y.W.L.) and Computer Science (H.S.W.), City University of Hong Kong, Hong Kong, P.R. China; Department of Computer Science, Guangzhou University, Guangzhou, P.R. China (S.Z.); Department of Bioengineering, Boston University, MA (A.C., C.C.); Harvard Wyss Institute for Biologically Inspired Engineering, Boston, MA (A.C., C.C.); Department of Cell Biology and Human Anatomy, University of California, Davis (J.M., A.H., D.K.L.); Cardiovascular Research Center, Mount Sinai School of Medicine, New York (D.K.L., R.A.L.); and Division of Cardiology, Johns Hopkins University, Baltimore, MD (G.F.T., K.R.B.).
authors.forename: Jennifer
authors.initials: J
authors.lastname: Moore
authors.affiliation: From the Stem Cell and Regenerative Medicine Consortium (E.P., W.K., B.Y., Z.W., O.T. W., K.R.B., R.A.L.) and Department of Physiology, LKS Faculty of Medicine (E.P., W.K., B.Y., Z.W., O.T.W., K.R.B., R.A.L.), University of Hong Kong, Hong Kong, P.R. China; Departments of Biology and Chemistry (Y.M.L., R.R., Y.W.L.) and Computer Science (H.S.W.), City University of Hong Kong, Hong Kong, P.R. China; Department of Computer Science, Guangzhou University, Guangzhou, P.R. China (S.Z.); Department of Bioengineering, Boston University, MA (A.C., C.C.); Harvard Wyss Institute for Biologically Inspired Engineering, Boston, MA (A.C., C.C.); Department of Cell Biology and Human Anatomy, University of California, Davis (J.M., A.H., D.K.L.); Cardiovascular Research Center, Mount Sinai School of Medicine, New York (D.K.L., R.A.L.); and Division of Cardiology, Johns Hopkins University, Baltimore, MD (G.F.T., K.R.B.).
authors.forename: Anthony
authors.initials: A
authors.lastname: Herren
authors.affiliation: From the Stem Cell and Regenerative Medicine Consortium (E.P., W.K., B.Y., Z.W., O.T. W., K.R.B., R.A.L.) and Department of Physiology, LKS Faculty of Medicine (E.P., W.K., B.Y., Z.W., O.T.W., K.R.B., R.A.L.), University of Hong Kong, Hong Kong, P.R. China; Departments of Biology and Chemistry (Y.M.L., R.R., Y.W.L.) and Computer Science (H.S.W.), City University of Hong Kong, Hong Kong, P.R. China; Department of Computer Science, Guangzhou University, Guangzhou, P.R. China (S.Z.); Department of Bioengineering, Boston University, MA (A.C., C.C.); Harvard Wyss Institute for Biologically Inspired Engineering, Boston, MA (A.C., C.C.); Department of Cell Biology and Human Anatomy, University of California, Davis (J.M., A.H., D.K.L.); Cardiovascular Research Center, Mount Sinai School of Medicine, New York (D.K.L., R.A.L.); and Division of Cardiology, Johns Hopkins University, Baltimore, MD (G.F.T., K.R.B.).
authors.forename: Deborah K
authors.initials: DK
authors.lastname: Lieu
authors.affiliation: From the Stem Cell and Regenerative Medicine Consortium (E.P., W.K., B.Y., Z.W., O.T. W., K.R.B., R.A.L.) and Department of Physiology, LKS Faculty of Medicine (E.P., W.K., B.Y., Z.W., O.T.W., K.R.B., R.A.L.), University of Hong Kong, Hong Kong, P.R. China; Departments of Biology and Chemistry (Y.M.L., R.R., Y.W.L.) and Computer Science (H.S.W.), City University of Hong Kong, Hong Kong, P.R. China; Department of Computer Science, Guangzhou University, Guangzhou, P.R. China (S.Z.); Department of Bioengineering, Boston University, MA (A.C., C.C.); Harvard Wyss Institute for Biologically Inspired Engineering, Boston, MA (A.C., C.C.); Department of Cell Biology and Human Anatomy, University of California, Davis (J.M., A.H., D.K.L.); Cardiovascular Research Center, Mount Sinai School of Medicine, New York (D.K.L., R.A.L.); and Division of Cardiology, Johns Hopkins University, Baltimore, MD (G.F.T., K.R.B.).
authors.forename: Hau San
authors.initials: HS
authors.lastname: Wong
authors.affiliation: From the Stem Cell and Regenerative Medicine Consortium (E.P., W.K., B.Y., Z.W., O.T. W., K.R.B., R.A.L.) and Department of Physiology, LKS Faculty of Medicine (E.P., W.K., B.Y., Z.W., O.T.W., K.R.B., R.A.L.), University of Hong Kong, Hong Kong, P.R. China; Departments of Biology and Chemistry (Y.M.L., R.R., Y.W.L.) and Computer Science (H.S.W.), City University of Hong Kong, Hong Kong, P.R. China; Department of Computer Science, Guangzhou University, Guangzhou, P.R. China (S.Z.); Department of Bioengineering, Boston University, MA (A.C., C.C.); Harvard Wyss Institute for Biologically Inspired Engineering, Boston, MA (A.C., C.C.); Department of Cell Biology and Human Anatomy, University of California, Davis (J.M., A.H., D.K.L.); Cardiovascular Research Center, Mount Sinai School of Medicine, New York (D.K.L., R.A.L.); and Division of Cardiology, Johns Hopkins University, Baltimore, MD (G.F.T., K.R.B.).
authors.forename: Zhihui
authors.initials: Z
authors.lastname: Weng
authors.affiliation: From the Stem Cell and Regenerative Medicine Consortium (E.P., W.K., B.Y., Z.W., O.T. W., K.R.B., R.A.L.) and Department of Physiology, LKS Faculty of Medicine (E.P., W.K., B.Y., Z.W., O.T.W., K.R.B., R.A.L.), University of Hong Kong, Hong Kong, P.R. China; Departments of Biology and Chemistry (Y.M.L., R.R., Y.W.L.) and Computer Science (H.S.W.), City University of Hong Kong, Hong Kong, P.R. China; Department of Computer Science, Guangzhou University, Guangzhou, P.R. China (S.Z.); Department of Bioengineering, Boston University, MA (A.C., C.C.); Harvard Wyss Institute for Biologically Inspired Engineering, Boston, MA (A.C., C.C.); Department of Cell Biology and Human Anatomy, University of California, Davis (J.M., A.H., D.K.L.); Cardiovascular Research Center, Mount Sinai School of Medicine, New York (D.K.L., R.A.L.); and Division of Cardiology, Johns Hopkins University, Baltimore, MD (G.F.T., K.R.B.).
authors.forename: On Tik
authors.initials: OT
authors.lastname: Wong
authors.affiliation: From the Stem Cell and Regenerative Medicine Consortium (E.P., W.K., B.Y., Z.W., O.T. W., K.R.B., R.A.L.) and Department of Physiology, LKS Faculty of Medicine (E.P., W.K., B.Y., Z.W., O.T.W., K.R.B., R.A.L.), University of Hong Kong, Hong Kong, P.R. China; Departments of Biology and Chemistry (Y.M.L., R.R., Y.W.L.) and Computer Science (H.S.W.), City University of Hong Kong, Hong Kong, P.R. China; Department of Computer Science, Guangzhou University, Guangzhou, P.R. China (S.Z.); Department of Bioengineering, Boston University, MA (A.C., C.C.); Harvard Wyss Institute for Biologically Inspired Engineering, Boston, MA (A.C., C.C.); Department of Cell Biology and Human Anatomy, University of California, Davis (J.M., A.H., D.K.L.); Cardiovascular Research Center, Mount Sinai School of Medicine, New York (D.K.L., R.A.L.); and Division of Cardiology, Johns Hopkins University, Baltimore, MD (G.F.T., K.R.B.).
authors.forename: Yun Wah
authors.initials: YW
authors.lastname: Lam
authors.affiliation: From the Stem Cell and Regenerative Medicine Consortium (E.P., W.K., B.Y., Z.W., O.T. W., K.R.B., R.A.L.) and Department of Physiology, LKS Faculty of Medicine (E.P., W.K., B.Y., Z.W., O.T.W., K.R.B., R.A.L.), University of Hong Kong, Hong Kong, P.R. China; Departments of Biology and Chemistry (Y.M.L., R.R., Y.W.L.) and Computer Science (H.S.W.), City University of Hong Kong, Hong Kong, P.R. China; Department of Computer Science, Guangzhou University, Guangzhou, P.R. China (S.Z.); Department of Bioengineering, Boston University, MA (A.C., C.C.); Harvard Wyss Institute for Biologically Inspired Engineering, Boston, MA (A.C., C.C.); Department of Cell Biology and Human Anatomy, University of California, Davis (J.M., A.H., D.K.L.); Cardiovascular Research Center, Mount Sinai School of Medicine, New York (D.K.L., R.A.L.); and Division of Cardiology, Johns Hopkins University, Baltimore, MD (G.F.T., K.R.B.).
authors.forename: Gordon F
authors.initials: GF
authors.lastname: Tomaselli
authors.affiliation: From the Stem Cell and Regenerative Medicine Consortium (E.P., W.K., B.Y., Z.W., O.T. W., K.R.B., R.A.L.) and Department of Physiology, LKS Faculty of Medicine (E.P., W.K., B.Y., Z.W., O.T.W., K.R.B., R.A.L.), University of Hong Kong, Hong Kong, P.R. China; Departments of Biology and Chemistry (Y.M.L., R.R., Y.W.L.) and Computer Science (H.S.W.), City University of Hong Kong, Hong Kong, P.R. China; Department of Computer Science, Guangzhou University, Guangzhou, P.R. China (S.Z.); Department of Bioengineering, Boston University, MA (A.C., C.C.); Harvard Wyss Institute for Biologically Inspired Engineering, Boston, MA (A.C., C.C.); Department of Cell Biology and Human Anatomy, University of California, Davis (J.M., A.H., D.K.L.); Cardiovascular Research Center, Mount Sinai School of Medicine, New York (D.K.L., R.A.L.); and Division of Cardiology, Johns Hopkins University, Baltimore, MD (G.F.T., K.R.B.).
authors.forename: Christopher
authors.initials: C
authors.lastname: Chen
authors.affiliation: From the Stem Cell and Regenerative Medicine Consortium (E.P., W.K., B.Y., Z.W., O.T. W., K.R.B., R.A.L.) and Department of Physiology, LKS Faculty of Medicine (E.P., W.K., B.Y., Z.W., O.T.W., K.R.B., R.A.L.), University of Hong Kong, Hong Kong, P.R. China; Departments of Biology and Chemistry (Y.M.L., R.R., Y.W.L.) and Computer Science (H.S.W.), City University of Hong Kong, Hong Kong, P.R. China; Department of Computer Science, Guangzhou University, Guangzhou, P.R. China (S.Z.); Department of Bioengineering, Boston University, MA (A.C., C.C.); Harvard Wyss Institute for Biologically Inspired Engineering, Boston, MA (A.C., C.C.); Department of Cell Biology and Human Anatomy, University of California, Davis (J.M., A.H., D.K.L.); Cardiovascular Research Center, Mount Sinai School of Medicine, New York (D.K.L., R.A.L.); and Division of Cardiology, Johns Hopkins University, Baltimore, MD (G.F.T., K.R.B.).
authors.forename: Kenneth R
authors.initials: KR
authors.lastname: Boheler
authors.affiliation: From the Stem Cell and Regenerative Medicine Consortium (E.P., W.K., B.Y., Z.W., O.T. W., K.R.B., R.A.L.) and Department of Physiology, LKS Faculty of Medicine (E.P., W.K., B.Y., Z.W., O.T.W., K.R.B., R.A.L.), University of Hong Kong, Hong Kong, P.R. China; Departments of Biology and Chemistry (Y.M.L., R.R., Y.W.L.) and Computer Science (H.S.W.), City University of Hong Kong, Hong Kong, P.R. China; Department of Computer Science, Guangzhou University, Guangzhou, P.R. China (S.Z.); Department of Bioengineering, Boston University, MA (A.C., C.C.); Harvard Wyss Institute for Biologically Inspired Engineering, Boston, MA (A.C., C.C.); Department of Cell Biology and Human Anatomy, University of California, Davis (J.M., A.H., D.K.L.); Cardiovascular Research Center, Mount Sinai School of Medicine, New York (D.K.L., R.A.L.); and Division of Cardiology, Johns Hopkins University, Baltimore, MD (G.F.T., K.R.B.). ronaldli@hku.hk.
authors.forename: Ronald A
authors.initials: RA
authors.lastname: Li
mesh_terms: D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D016000:Cluster Analysis; D015180:Electrophoresis, Gel, Two-Dimensional; D005333:Fetus / Q000166:cytology*; D005786:Gene Expression Regulation / Q000187:drug effects; D006352:Heart Ventricles / Q000166:cytology / Q000378:metabolism; D000066449:Human Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D015227:Lipid Peroxidation / Q000187:drug effects; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D009119:Muscle Contraction / Q000187:drug effects; D009206:Myocardium / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000166:cytology*; D047493:PPAR alpha / Q000037:antagonists & inhibitors / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D040901:Proteomics; D011743:Pyrimidines / Q000494:pharmacology; D014284:Triiodothyronine / Q000494:pharmacology
chemical_list: D047493:PPAR alpha; D011743:Pyrimidines; D014284:Triiodothyronine; C006253:pirinixic acid
keywords: PGC1alpha protein; PPARalpha; embryonic stem cells; metabolism; proteomics; thyroid-stimulating hormone