Interaction of Wnt/β-catenin and notch signaling in the early stage of cardiac differentiation of P19CL6 cells

2012 ◽  
Vol 113 (2) ◽  
pp. 629-639 ◽  
Author(s):  
Binhong Li ◽  
Zhuqing Jia ◽  
Tao Wang ◽  
Weiping Wang ◽  
Chenguang Zhang ◽  
...  
Keyword(s):  
Notch Signaling ◽  
Early Stage ◽  
Cardiac Differentiation ◽  
P19cl6 Cells

scholarly journals Rapamycin efficiently promotes cardiac differentiation of mouse embryonic stem cells

2017 ◽  
Vol 37 (3) ◽  
Author(s):  
Qin Lu ◽  
Yinan Liu ◽  
Yang Wang ◽  
Weiping Wang ◽  
Zhe Yang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Early Stage ◽  
Mammalian Target Of Rapamycin ◽  
Embryonic Stem ◽  
Cardiac Differentiation ◽  
Bone Morphogenetic Protein 2 ◽  
Morphogenetic Protein ◽  
Cardiac Transcription Factors ◽  
20 Nm

To investigate the effects of rapamycin on cardiac differentiation, murine embryonic stem cells (ESCs) were induced into cardiomyocytes by 10−4 M ascorbic acid (AA), 20 nM rapamycin alone or 0.01% solvent DMSO. We found that rapamycin alone was insufficient to initiate cardiomyogenesis. Then, the ESCs were treated with AA and rapamycin (20 nM) or AA and DMSO (0.01%) as a control. Compared with control, mouse ESCs (mESCs) treated with rapamycin (20 nM) and AA yielded a significantly higher percentage of cardiomyocytes, as confirmed by the percentage of beating embryonic bodies (EBs), the immunofluorescence and FACS analysis. Rapamycin significantly increased the expression of a panel of cardiac markers including Gata4, α-Mhc, β-Mhc, and Tnnt2. Additionally, rapamycin enhanced the expression of mesodermal and cardiac transcription factors such as Mesp1, Brachyury T, Eomes, Isl1, Gata4, Nkx2.5, Tbx5, and Mef2c. Mechanistic studies showed that rapamycin inhibits Wnt/β-catenin and Notch signaling but promotes the expression of fibroblast growth factor (Fgf8), Fgf10, and Nodal at early stage, and bone morphogenetic protein 2 (Bmp 2) at later stages. Sequential treatment of rapamycin showed that rapamycin promotes cardiac differentiation at the early and later stages. Interestingly, another mammalian target of rapamycin (mTOR) inhibitor Ku0063794 (1 µM) had similar effects on cardiomyogenesis. In conclusion, our results highlight a practical approach to generate cardiomyocytes from mESCs by rapamycin.

scholarly journals 4-phenylbutyrate exerts stage-specific effects on cardiac differentiation via HDAC inhibition

PLoS ONE ◽  
2021 ◽  
Vol 16 (4) ◽  
pp. e0250267
Author(s):  
Yanming Li ◽  
Xiaofei Weng ◽  
Pingping Wang ◽  
Zezhao He ◽  
Siya Cheng ◽  
...  
Keyword(s):  
Late Stage ◽  
Histone Deacetylases ◽  
Early Stage ◽  
Trichostatin A ◽  
Es Cells ◽  
Hdac Inhibitors ◽  
Embryonic Stem ◽  
Cardiac Differentiation ◽  
Temporal Expression ◽  
Cardiac Progenitors

4-phenylbutyrate (4-PBA), a terminal aromatic substituted fatty acid, is used widely to specifically attenuate endoplasmic reticulum (ER) stress and inhibit histone deacetylases (HDACs). In this study, we investigated the effect of 4-PBA on cardiac differentiation of mouse embryonic stem (ES) cells. Herein, we found that 4-PBA regulated cardiac differentiation in a stage-specific manner just like trichostatin A (TSA), a well-known HDAC inhibitor. 4-PBA and TSA favored the early-stage differentiation, but inhibited the late-stage cardiac differentiation via acetylation. Mechanistic studies suggested that HDACs exhibited a temporal expression profiling during cardiomyogenesis. Hdac1 expression underwent a decrease at the early stage, while was upregulated at the late stage of cardiac induction. During the early stage of cardiac differentiation, acetylation favored the induction of Isl1 and Nkx2.5, two transcription factors of cardiac progenitors. During the late stage, histone acetylation induced by 4-PBA or TSA interrupted the gene silence of Oct4, a key determinant of self-renewal and pluripotency. Thereby, 4-PBA and TSA at the late stage hindered the exit from pluripotency, and attenuated the expression of cardiac-specific contractile proteins. Overexpression of HDAC1 and p300 exerted different effects at the distinct stages of cardiac induction. Collectively, our study shows that timely manipulation of HDACs exhibits distinct effects on cardiac differentiation. And the context-dependent effects of HDAC inhibitors depend on cell differentiation states marked by the temporal expression of pluripotency-associated genes.

Abstract 1964: Bone Morphogenetic Protein Antagonist “Protein Related to Dan and Cerberus” Promotes Differentiation of Embryonic Stem Cells to Atrial Cardiomyocytes

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Jingbo Yan ◽  
Jianyong Hu ◽  
Iris I Mueller ◽  
William H Heaton ◽  
Wan-Der Wang ◽  
...  
Keyword(s):  
Notch Signaling ◽  
Progenitor Cells ◽  
Molecular Mechanisms ◽  
Es Cells ◽  
Embryonic Stem ◽  
Cell Types ◽  
Bmp Signaling ◽  
Cardiac Differentiation ◽  
Cardiovascular Cells

The molecular factors that regulate cardiac differentiation have been extensively studied, yet, relatively little is known about how cardiomyocytes acquire atrial versus ventricular characteristics. Embryonic stem (ES) cells, which have the potential to differentiate to a wide array of distinct cell types, including most types of cardiovascular cells, offer a pertinent in vitro model to work out the molecular mechanisms of atrial specification and differentiation. We discovered that the secreted antagonist of BMP signaling, Protein Related to Dan and Cerberus (PRDC, also called Gremlin2) leads to a surge in cardiomyocytic differentiation when applied to mouse ES-derived cardiac progenitor cells. This property is unique to PRDC among tested BMP antagonists. Lineage expansion is restricted to cardiomyocytes, with the differentiation of endodermal, blood, endothelial and neuronal cells being unaffected. Using molecular and electrophysiological analyses, we show that PRDC-induced cardiomyocytes acquire atrial characteristics. Consistent with the in vitro results, we found that injection of PRDC mRNA into the developing zebrafish embryo leads to supernumerary contracting areas. The ectopic cardiomyocytes express atrial-, but not ventricular- specific cardiac genes. We determined that PRDC treatment induces the expression of COUP-TFII, a known transcriptional regulator of atrial differentiation, but suppresses Notch signaling. Inhibition of Notch is sufficient to induce atrial-specific genes; however, blocking Notch does not expand the cardiogenic fields. Taken together, our data suggest that antagonism of BMP and Notch signaling by PRDC is a critical early step in the specification, expansion and differentiation of atrial progenitor cells. This information might be relevant for treating atrial degeneration, as well as for understanding the etiology of atrial fibrillation.

Abstract 689: Notch Signaling Stimulates Expression Of Wnts And Contributes To Cardiac Markers Gene Expression In Human Progenitor Cell

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Masamichi Koyanagi ◽  
Philipp Bushoven ◽  
Masayoshi Iwasaki ◽  
Carmen Urbich ◽  
Andreas M Zeiher ◽  
...  
Keyword(s):  
Gene Expression ◽  
Notch Signaling ◽  
Human Cells ◽  
Cardiac Differentiation ◽  
Neonatal Rat ◽  
Notch 1 ◽  
Cardiac Marker ◽  
Wnt Proteins ◽  
Mesenchymal Transformation ◽  
Notch Activation

It has been demonstrated that adult human circulating endothelial progenitor cells (EPC) can differentiate to a cardiomyogenic phenotype. Notch signaling promotes epithelial-to-mesenchymal transformation and plays a prominent role in heart and vessel development. Here, we investigated the role of Notch activation for cardiac differentiation of EPC in a co-culture system with neonatal rat cardiomyocyte (CM). EPC expressed the receptors Notch-1 and Notch-2, whereas CM expressed high level of Notch ligand Jagged-1. Therefore, we hypothesized that CM may activate Notch signaling within EPC. Indeed, after co-culture, Notch activation was detected in EPC by immunohistochemical detection of the intracellular cleavage fragment of Notch-1 (NICD), whereas NICD was only rarely detected in EPC before co-culture. Western blot analysis confirmed Notch cleavage after co-culture. RT-PCR directed against the human specific sequences of the Notch target genes Hey2 and Hes1 demonstrated a transient activation of the transcriptional activity of Notch in human EPC after co-culture with CM. Inhibition of γ-secretase significantly blocked Notch cleavages and NICD translocations. Furthermore, the expression of the cardiac marker protein γ-sarcomeric actinin and troponin T was significantly suppressed by γ-secretase inhibition (55.8 ± 8.1% and 54.0 ± 10.5% of control, respectively) or addition of soluble recombinant Jagged-1, indicating that Notch activation facilitates cardiac marker gene expression. Because non-canonical Wnts have previously been shown to promote cardiac differentiation, we additionally determined the influence of Notch activation on the expression of Wnt5a and. Wnt5a and Wnt11 expressions in the human cells was induced by the co-culture and was blocked by γ-secretase inhibitor. Likewise, stimulation of Notch signaling by immobilized Jagged-1 promoted NICD cleavage and Wnt5a expression in EPC. These data suggested that Notch is transiently activated upon co-culture of EPC with neonatal rat CM. γ-Secretase-dependent Notch activation is required for cardiac gene expression in human cells and induces the expression of non-canonical Wnt proteins, which may act in an paracrine manner to further amplify cardiac differentiation.

POU homeodomain protein OCT1 modulates islet 1 expression during cardiac differentiation of P19CL6 cells

2010 ◽  
Vol 68 (11) ◽  
pp. 1969-1982 ◽  
Author(s):  
Yinan Liu ◽  
Yanming Li ◽  
Tao Li ◽  
Huafei Lu ◽  
Zhuqing Jia ◽  
...  
Keyword(s):  
Cardiac Differentiation ◽  
Homeodomain Protein ◽  
P19cl6 Cells ◽  
Islet 1

scholarly journals Dynamic regulation of small RNAome during the early stage of cardiac differentiation from pluripotent embryonic stem cells

Genomics Data ◽  
2017 ◽  
Vol 12 ◽  
pp. 136-145 ◽  
Author(s):  
Yue Li ◽  
An Zeng ◽  
Ge Li ◽  
Ya-Na Guan ◽  
Huang-Tian Yang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Early Stage ◽  
Embryonic Stem ◽  
Cardiac Differentiation ◽  
Dynamic Regulation

scholarly journals Insulin induces proliferation and cardiac differentiation of P19CL6 cells in a dose-dependent manner

2013 ◽  
Vol 55 (7) ◽  
pp. 676-686 ◽  
Author(s):  
Wen-Yan Li ◽  
Yang-Liu Song ◽  
Cheng-Juan Xiong ◽  
Pei-Qi Lu ◽  
Li-Xiang Xue ◽  
...  
Keyword(s):  
Cardiac Differentiation ◽  
Dependent Manner ◽  
P19cl6 Cells ◽  
Dose Dependent

P3479Sequential stimulation and inhibition of lysophosphatidic acid receptor 4 are critical for cardiac differentiation and repair

2019 ◽  
Vol 40 (Supplement_1) ◽  
Author(s):  
H J Cho ◽  
J W Lee ◽  
C S Lee ◽  
Y R Ryu ◽  
H S Kim ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Surface ◽  
Progenitor Cells ◽  
Lysophosphatidic Acid ◽  
Functional Significance ◽  
Early Stage ◽  
Cardiac Differentiation ◽  
Cardiac Progenitor Cells ◽  
Acid Receptor ◽  
Downstream Signaling

Abstract Background The clinical application of cell therapy to repair the damaged heart needs to understand the precise differentiation process of stem cells and the characteristics of cardiac progenitor cells. Purpose We examined the cardiac-specific markers that expressed on the cell surface and determined their functional significance during cardiac differentiation. Methods and results We screened cell-surface expressing proteins on cardiac progenitor cells at differentiation day 3 compared to undifferentiated pluripotent stem cells (PSCs). Among candidates, we identified lysophosphatidic acid receptor 4 (LPAR4) that is a G protein-coupled receptor. During in vitro differentiation of mouse PSCs toward cardiac cells, LPAR4 expression peaked for 3–5 days and then and declined immediately. Also in vivo, LPAR4 was specifically expressed in the early stage of heart development in embryos and disappeared completely in adults, suggesting that stimulatory signal of LPAR4 at an early stage should be shut off for further progression of differentiation. We next have identified the LPAR4 downstream signaling molecule, p38MAPK, by comparing PSCs and LPAR4 knockdown PSCs. In both mouse and human PSCs, ODP (LPAR4 specific agonist) followed by p38MAPK blocker (SB203580) treatment significantly increased cardiac differentiation efficiency. Furthermore, we investigated whether LPAR4 is the maker for adult cardiac progenitor cells. We found that LPAR4-positive cells were rarely present in normal adult mouse hearts, but LPAR4-positive cells were increased when the heart was damaged. LPAR4-positive cells from adult hearts differentiated into cardiomyocytes. After myocardial infarction (MI), the sequential stimulation and inhibition of LPAR4 with ODP and p38MAPK blocker resulted in the reduction of infarct size and improvement of left ventricular dysfunction. Conclusion We demonstrated that LPAR4 is a cardiac progenitor-specific marker and its functional significance during cardiac differentiation and regeneration. Our findings provide a new insight in cell-free cardiac repair by the modulation of progenitor-specific downstream signaling. Acknowledgement/Funding Grants from “Strategic Center of Cell and Bio Therapy” (grant number: HI17C2085) and “Korea Research-Driven Hospital” (HI14C1277)

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