Click
here to close Hello! We notice that
you are using Internet Explorer, which is not supported by Echinobase
and may cause the site to display incorrectly. We suggest using a
current version of Chrome,
FireFox,
or Safari.
Mar Drugs
2018 Jun 02;166:. doi: 10.3390/md16060192.
Show Gene links
Show Anatomy links
A Novel Atypical PKC-Iota Inhibitor, Echinochrome A, Enhances Cardiomyocyte Differentiation from Mouse Embryonic Stem Cells.
Kim HK
,
Cho SW
,
Heo HJ
,
Jeong SH
,
Kim M
,
Ko KS
,
Rhee BD
,
Mishchenko NP
,
Vasileva EA
,
Fedoreyev SA
,
Stonik VA
,
Han J
.
???displayArticle.abstract???
Echinochrome A (EchA) is a marine bioproduct extracted from sea urchins having antioxidant, antimicrobial, anti-inflammatory, and chelating effects, and is the active component of the clinical drug histochrome. We investigated the potential use of Ech A for inducing cardiomyocyte differentiation from mouse embryonic stem cells (mESCs). We also assessed the effects of Ech A on mitochondrial mass, inner membrane potential (Δψm), reactive oxygen species generation, and levels of Ca2+. To identify the direct target of Ech A, we performed in vitro kinase activity and surface plasmon resonance binding assays. Ech A dose-dependently enhanced cardiomyocyte differentiation with higher beating rates. Ech A (50 μM) increased the mitochondrial mass and membrane potential but did not alter the mitochondrial superoxide and Ca2+ levels. The in vitro kinase activity of the atypical protein kinase C-iota (PKCι) was significantly decreased by 50 μM of Ech A with an IC50 for PKCι activity of 107 μM. Computational protein-ligand docking simulation results suggested the direct binding of Ech A to PKCι, and surface plasmon resonance confirmed the direct binding with a low KD of 6.3 nM. Therefore, Ech A is a potential drug for enhancing cardiomyocyte differentiation from mESCs through direct binding to PKCι and inhibition of its activity.
Figure 1. The chemical structure and cell viability effect of echinochrome A (Ech A). (A) The chemical structure of Ech A (6-ethyl-2,3,5,7,8-pentahydroxy-1,4-naphthoquinone; FW = 266.2). (B) The cell viability according to the dose of Ech A. Each group, n = 3–5.
Figure 2. Ech A enhances cardiomyocyte differentiation from mouse embryonic stem cells (mESCs). (A) The protocol for the differentiation of cardiomyocytes from the mESCs using the hanging drop method to produce differentiated embryoid bodies (EBs). (B) The beating number per minute (BPM) of EBs during differentiation. Each group, n = 3. * p < 0.05 (C) Representative FACS analyses and the percentage of mESC-derived αMHC+ cardiomyocyte-like cells according to the dose of Ech A. Each group, n = 3. (D) Relative mRNA expression levels of cardiomyocyte specific genes in nontreated control (Con), Ech A treatment (Ech), heart tissue from neonatal (7 day, Neo) or adult mice (10 weeks, Adult). Each group, n = 3. * p < 0.05 vs. Con, †p < 0.05 vs. Ech A. The mRNA level of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as an internal control. (E) Western blot analyses for the expression of cardiomyocyte specific proteins incubated with the control vehicle (Con) or 50 µM of Ech A. (F) Images displaying αMHC-GFP+, α-actinin+, cTnT+, and Cx43+ cells incubated with Con or 50 µM of Ech A (scale bars, 50 μm). SCC: side scatter cell, cTnI: cardiac troponin I, cTNT: cardiac troponin T, Cx43: connexin 43, Mef2c: Myocyte Enhancer Factor 2C, Mlc2: Myosin light chain 2.
Figure 5. Computer simulated Ech A-PKCι docking model. (A) A three-dimensional (3D) model of Ech A-PKCι (PDB:3A8W) docking simulation was generated by using the online DockingServer site (https://www.dockingserver.com). (B) Magnified 3D structure of the Ech A-PKCι binding site.
Figure 6. In vitro surface plasmon resonance binding assay for PKCι and Ech A. The PKCι binding response to Ech A was measured in the range of 0–40 nM of Ech A. The flow rate was 30 μL/minute. The association and dissociation times were 3 min each. The calculated binding constants are shown at the bottom. ka: Association rate, kd: Dissociation rate, KD (Affinity) = kd/ka (n = 3).
Ali,
A novel adamantane thiadiazole derivative induces mitochondria-mediated apoptosis in lung carcinoma cell line.
2017, Pubmed
Ali,
A novel adamantane thiadiazole derivative induces mitochondria-mediated apoptosis in lung carcinoma cell line.
2017,
Pubmed
Bikadi,
Application of the PM6 semi-empirical method to modeling proteins enhances docking accuracy of AutoDock.
2009,
Pubmed
Burridge,
Production of de novo cardiomyocytes: human pluripotent stem cell differentiation and direct reprogramming.
2012,
Pubmed
Butler,
A small molecule inhibitor of atypical protein kinase C signaling inhibits pancreatic cancer cell transformed growth and invasion.
2015,
Pubmed
Buĭmov,
[Effect of the bioantioxidant histochrome on myocardial injury in reperfusion therapy on patients with myocardial infarction].
2002,
Pubmed
Chand,
Protein kinase C-theta inhibitors: a novel therapy for inflammatory disorders.
2012,
Pubmed
Cho,
Dual modulation of the mitochondrial permeability transition pore and redox signaling synergistically promotes cardiomyocyte differentiation from pluripotent stem cells.
2014,
Pubmed
Chung,
Mitochondrial oxidative metabolism is required for the cardiac differentiation of stem cells.
2007,
Pubmed
Desai,
Regulation of Cdk7 activity through a phosphatidylinositol (3)-kinase/PKC-ι-mediated signaling cascade in glioblastoma.
2012,
Pubmed
Hansson,
Regeneration next: toward heart stem cell therapeutics.
2009,
Pubmed
Heo,
Mitochondrial pyruvate dehydrogenase phosphatase 1 regulates the early differentiation of cardiomyocytes from mouse embryonic stem cells.
2016,
Pubmed
Hong,
Generation of PDGFRα+ Cardioblasts from Pluripotent Stem Cells.
2017,
Pubmed
Jatoi,
A proof-of-concept trial of protein kinase C iota inhibition with auranofin for the paclitaxel-induced acute pain syndrome.
2017,
Pubmed
Jeong,
Echinochrome a increases mitochondrial mass and function by modulating mitochondrial biogenesis regulatory genes.
2014,
Pubmed
,
Echinobase
Jeong,
Echinochrome A protects mitochondrial function in cardiomyocytes against cardiotoxic drugs.
2014,
Pubmed
,
Echinobase
Kim,
Echinochrome A regulates phosphorylation of phospholamban Ser16 and Thr17 suppressing cardiac SERCA2A Ca²⁺ reuptake.
2015,
Pubmed
,
Echinobase
Lebedev,
Echinochrome, a naturally occurring iron chelator and free radical scavenger in artificial and natural membrane systems.
2005,
Pubmed
Lin,
Strategies for cardiac regeneration and repair.
2014,
Pubmed
Maculins,
A Generic Platform for Cellular Screening Against Ubiquitin Ligases.
2016,
Pubmed
Mah,
Atypical PKC-iota Controls Stem Cell Expansion via Regulation of the Notch Pathway.
2015,
Pubmed
Mahato,
Regulation of mitochondrial function and cellular energy metabolism by protein kinase C-λ/ι: a novel mode of balancing pluripotency.
2014,
Pubmed
McCray,
The interruption of PKC-ι signaling and TRAIL combination therapy against glioblastoma cells.
2014,
Pubmed
Passier,
Stem-cell-based therapy and lessons from the heart.
2008,
Pubmed
Pillai,
A novel PKC-ι inhibitor abrogates cell proliferation and induces apoptosis in neuroblastoma.
2011,
Pubmed
Porter,
Bioenergetics, mitochondria, and cardiac myocyte differentiation.
2011,
Pubmed
Ratnayake,
Two novel atypical PKC inhibitors; ACPD and DNDA effectively mitigate cell proliferation and epithelial to mesenchymal transition of metastatic melanoma while inducing apoptosis.
2017,
Pubmed
Segers,
Stem-cell therapy for cardiac disease.
2008,
Pubmed
Seidl,
Phenotypical analysis of atypical PKCs in vivo function display a compensatory system at mouse embryonic day 7.5.
2013,
Pubmed
Serasinghe,
Identification and characterization of unique proline-rich peptides binding to the mitochondrial fission protein hFis1.
2010,
Pubmed
Shiba,
Allogeneic transplantation of iPS cell-derived cardiomyocytes regenerates primate hearts.
2016,
Pubmed
Shvilkin,
[Effect of echinochrom on experimental myocardial reperfusion injury].
1991,
Pubmed
Spitkovsky,
Activity of complex III of the mitochondrial electron transport chain is essential for early heart muscle cell differentiation.
2004,
Pubmed
Thomson,
Distribution of naturally occurring quinones.
1991,
Pubmed
Tohyama,
Distinct metabolic flow enables large-scale purification of mouse and human pluripotent stem cell-derived cardiomyocytes.
2013,
Pubmed
Win,
Role of protein kinase C-iota in transformed non-malignant RWPE-1 cells and androgen-independent prostate carcinoma DU-145 cells.
2009,
Pubmed
