PMID- 31707831
OWN - NLM
STAT- MEDLINE
DCOM- 20200630
LR  - 20230729
IS  - 1524-4539 (Electronic)
IS  - 0009-7322 (Print)
IS  - 0009-7322 (Linking)
VI  - 141
IP  - 4
DP  - 2020 Jan 28
TI  - Inhibition of mTOR Signaling Enhances Maturation of Cardiomyocytes Derived From 
      Human-Induced Pluripotent Stem Cells via p53-Induced Quiescence.
PG  - 285-300
LID - 10.1161/CIRCULATIONAHA.119.044205 [doi]
AB  - BACKGROUND: Current differentiation protocols to produce cardiomyocytes from 
      human induced pluripotent stem cells (iPSCs) are capable of generating highly 
      pure cardiomyocyte populations as determined by expression of cardiac troponin T. 
      However, these cardiomyocytes remain immature, more closely resembling the fetal 
      state, with a lower maximum contractile force, slower upstroke velocity, and 
      immature mitochondrial function compared with adult cardiomyocytes. Immaturity of 
      iPSC-derived cardiomyocytes may be a significant barrier to clinical translation 
      of cardiomyocyte cell therapies for heart disease. During development, 
      cardiomyocytes undergo a shift from a proliferative state in the fetus to a more 
      mature but quiescent state after birth. The mechanistic target of rapamycin 
      (mTOR)-signaling pathway plays a key role in nutrient sensing and growth. We 
      hypothesized that transient inhibition of the mTOR-signaling pathway could lead 
      cardiomyocytes to a quiescent state and enhance cardiomyocyte maturation. 
      METHODS: Cardiomyocytes were differentiated from 3 human iPSC lines using small 
      molecules to modulate the Wnt pathway. Torin1 (0 to 200 nmol/L) was used to 
      inhibit the mTOR pathway at various time points. We quantified contractile, 
      metabolic, and electrophysiological properties of matured iPSC-derived 
      cardiomyocytes. We utilized the small molecule inhibitor, pifithrin-alpha, to inhibit 
      p53 signaling, and nutlin-3a, a small molecule inhibitor of MDM2 (mouse double 
      minute 2 homolog) to upregulate and increase activation of p53. RESULTS: Torin1 
      (200 nmol/L) increased the percentage of quiescent cells (G(0) phase) from 24% to 
      48% compared with vehicle control (P<0.05). Torin1 significantly increased 
      expression of selected sarcomere proteins (including TNNI3 [troponin I, cardiac 
      muscle]) and ion channels (including Kir2.1) in a dose-dependent manner when 
      Torin1 was initiated after onset of cardiomyocyte beating. Torin1-treated cells 
      had an increased relative maximum force of contraction, increased maximum oxygen 
      consumption rate, decreased peak rise time, and increased downstroke velocity. 
      Torin1 treatment increased protein expression of p53, and these effects were 
      inhibited by pifithrin-alpha. In contrast, nutlin-3a independently upregulated p53, 
      led to an increase in TNNI3 expression and worked synergistically with Torin1 to 
      further increase expression of both p53 and TNNI3. CONCLUSIONS: Transient 
      treatment of human iPSC-derived cardiomyocytes with Torin1 shifts cells to a 
      quiescent state and enhances cardiomyocyte maturity.
FAU - Garbern, Jessica C
AU  - Garbern JC
AD  - Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell 
      Institute (J.C.G., A.H., R.S., M.S., A.A., G.O.E., D.A.M., R.T.L.), Harvard 
      University, Cambridge, MA.
AD  - Department of Cardiology, Boston Children's Hospital, MA (J.C.G., R.O., V.J.B.).
FAU - Helman, Aharon
AU  - Helman A
AD  - Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell 
      Institute (J.C.G., A.H., R.S., M.S., A.A., G.O.E., D.A.M., R.T.L.), Harvard 
      University, Cambridge, MA.
FAU - Sereda, Rebecca
AU  - Sereda R
AD  - Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell 
      Institute (J.C.G., A.H., R.S., M.S., A.A., G.O.E., D.A.M., R.T.L.), Harvard 
      University, Cambridge, MA.
FAU - Sarikhani, Mohsen
AU  - Sarikhani M
AD  - Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell 
      Institute (J.C.G., A.H., R.S., M.S., A.A., G.O.E., D.A.M., R.T.L.), Harvard 
      University, Cambridge, MA.
FAU - Ahmed, Aishah
AU  - Ahmed A
AD  - Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell 
      Institute (J.C.G., A.H., R.S., M.S., A.A., G.O.E., D.A.M., R.T.L.), Harvard 
      University, Cambridge, MA.
FAU - Escalante, Gabriela O
AU  - Escalante GO
AD  - Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell 
      Institute (J.C.G., A.H., R.S., M.S., A.A., G.O.E., D.A.M., R.T.L.), Harvard 
      University, Cambridge, MA.
FAU - Ogurlu, Roza
AU  - Ogurlu R
AD  - Department of Cardiology, Boston Children's Hospital, MA (J.C.G., R.O., V.J.B.).
FAU - Kim, Sean L
AU  - Kim SL
AD  - Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell 
      Institute (J.C.G., A.H., R.S., M.S., A.A., G.O.E., D.A.M., R.T.L.), Harvard 
      University, Cambridge, MA.
AD  - Disease Biophysics Group, Wyss Institute for Biologically Inspired Engineering, 
      School of Engineering and Applied Sciences (S.L.K., J.F.Z., A.C., L.M., K.K.P.), 
      Harvard University, Cambridge, MA.
FAU - Zimmerman, John F
AU  - Zimmerman JF
AD  - Disease Biophysics Group, Wyss Institute for Biologically Inspired Engineering, 
      School of Engineering and Applied Sciences (S.L.K., J.F.Z., A.C., L.M., K.K.P.), 
      Harvard University, Cambridge, MA.
FAU - Cho, Alexander
AU  - Cho A
AD  - Disease Biophysics Group, Wyss Institute for Biologically Inspired Engineering, 
      School of Engineering and Applied Sciences (S.L.K., J.F.Z., A.C., L.M., K.K.P.), 
      Harvard University, Cambridge, MA.
FAU - MacQueen, Luke
AU  - MacQueen L
AD  - Disease Biophysics Group, Wyss Institute for Biologically Inspired Engineering, 
      School of Engineering and Applied Sciences (S.L.K., J.F.Z., A.C., L.M., K.K.P.), 
      Harvard University, Cambridge, MA.
FAU - Bezzerides, Vassilios J
AU  - Bezzerides VJ
AD  - Department of Cardiology, Boston Children's Hospital, MA (J.C.G., R.O., V.J.B.).
FAU - Parker, Kevin Kit
AU  - Parker KK
AD  - Disease Biophysics Group, Wyss Institute for Biologically Inspired Engineering, 
      School of Engineering and Applied Sciences (S.L.K., J.F.Z., A.C., L.M., K.K.P.), 
      Harvard University, Cambridge, MA.
FAU - Melton, Douglas A
AU  - Melton DA
AD  - Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell 
      Institute (J.C.G., A.H., R.S., M.S., A.A., G.O.E., D.A.M., R.T.L.), Harvard 
      University, Cambridge, MA.
FAU - Lee, Richard T
AU  - Lee RT
AD  - Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell 
      Institute (J.C.G., A.H., R.S., M.S., A.A., G.O.E., D.A.M., R.T.L.), Harvard 
      University, Cambridge, MA.
AD  - Division of Cardiovascular Medicine, Department of Medicine, Brigham and Women's 
      Hospital and Harvard Medical School, Boston, MA (R.T.L.).
LA  - eng
GR  - UG3 HL141798/HL/NHLBI NIH HHS/United States
GR  - R01 HL151684/HL/NHLBI NIH HHS/United States
GR  - T32 HL007572/HL/NHLBI NIH HHS/United States
GR  - R01 HL137710/HL/NHLBI NIH HHS/United States
GR  - R01 HL119230/HL/NHLBI NIH HHS/United States
GR  - K08 HL140197/HL/NHLBI NIH HHS/United States
GR  - UH3 TR000522/TR/NCATS NIH HHS/United States
GR  - T32 EB016652/EB/NIBIB NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20191111
PL  - United States
TA  - Circulation
JT  - Circulation
JID - 0147763
RN  - 0 
      (1-(4-(4-propionylpiperazin-1-yl)-3-(trifluoromethyl)phenyl)-9-(quinolin-3-yl)benzo(h)(1,6)naphthyridin-2(1H)-one)
RN  - 0 (Benzothiazoles)
RN  - 0 (Imidazoles)
RN  - 0 (Naphthyridines)
RN  - 0 (Piperazines)
RN  - 0 (TP53 protein, human)
RN  - 0 (Tumor Suppressor Protein p53)
RN  - 3FPU23BG52 (Toluene)
RN  - 53IA0V845C (nutlin 3)
RN  - D213B92S1Y (pifithrin)
RN  - EC 2.7.1.1 (MTOR protein, human)
RN  - EC 2.7.11.1 (TOR Serine-Threonine Kinases)
SB  - IM
MH  - Benzothiazoles/pharmacology
MH  - Cell Line
MH  - Humans
MH  - Imidazoles/pharmacology
MH  - Induced Pluripotent Stem Cells/cytology/*metabolism
MH  - Myocytes, Cardiac/cytology/*metabolism
MH  - Naphthyridines/*pharmacology
MH  - Piperazines/pharmacology
MH  - TOR Serine-Threonine Kinases/antagonists & inhibitors/*metabolism
MH  - Toluene/analogs & derivatives/pharmacology
MH  - Tumor Suppressor Protein p53/antagonists & inhibitors/*metabolism
MH  - Wnt Signaling Pathway/*drug effects
PMC - PMC7009740
MID - NIHMS1549573
OTO - NOTNLM
OT  - MTOR protein
OT  - induced pluripotent stem cells
OT  - myocytes, cardiac
OT  - stem cells
OT  - troponin I
EDAT- 2019/11/12 06:00
MHDA- 2020/07/01 06:00
PMCR- 2021/01/28
CRDT- 2019/11/12 06:00
PHST- 2019/11/12 06:00 [pubmed]
PHST- 2020/07/01 06:00 [medline]
PHST- 2019/11/12 06:00 [entrez]
PHST- 2021/01/28 00:00 [pmc-release]
AID - 10.1161/CIRCULATIONAHA.119.044205 [doi]
PST - ppublish
SO  - Circulation. 2020 Jan 28;141(4):285-300. doi: 10.1161/CIRCULATIONAHA.119.044205. 
      Epub 2019 Nov 11.