Abstract 394: Transient Bone Morphogenic Protein Antagonism Directs Differentiation of iPSCs into the Cardiac Neural Crest and cKit+ Myocardial Progenitor Lineages

2015 ◽  
Vol 117 (suppl_1) ◽  
Author(s):  
Konstantinos E Hatzistergos ◽  
Lauro M Takeuchi ◽  
Dieter Saur ◽  
Barbara Seidler ◽  
Susan M Dymecki ◽  
...  
Keyword(s):  
Neural Crest ◽  
Signaling Pathway ◽  
Embryoid Bodies ◽  
Cardiomyocyte Differentiation ◽  
Cardiac Neural Crest ◽  
Specific Induction ◽  
Cre Recombination ◽  
Bmp Antagonist ◽  
Cardiogenic Differentiation ◽  
Cardiac Mesoderm

Introduction: The capability of cKit+ cardiac progenitor cells (CPCs) to participate in cardiomyocyte regeneration remains controversial, despite basic and clinical studies supporting such a role. Hypothesis: A non-permissive cardiac milieu minimizes the generation of cardiomyocytes from CPCs. Methods: We lineage-traced CPCs using novel dual-recombinase responsive indicator mice (cKitCreERT2;Wnt1::Flpe;RC::Fela) and iPSCs derived from cKitCreERT2;IRG (iPSCKit) mice. Results: Intersectional genetic fate-mapping of cKitCreERT2;Wnt1:: Flpe;RC::Fela embryos supported that cKit marks Wnt1-expressing cardiac neural crest (CNC) progenitors, emerging at ~E9.5 and contributing a limited number of cardiomyocytes. To decipher the mechanisms underlying cardiomyocyte differentiation of CPCs, we lineage-traced CPCs during stage-specific cardiogenic differentiation of iPSCKit. Ascorbate treatment promoted differentiation of cKit+ iPSC-derived embryoid bodies (EBs) into Nkx2.5+ myocardium, 45.5%±6.7% of which co-expressed the Cre-reporter EGFP (n=154 EBs; 12 preparations), suggesting that CPCs encompass fully competent cardiomyogenic progenitors. Noggin (or Dorsomorphin), a BMP antagonist transiently expressed in the heart at E7.5-E8.5 but not during CNC invasion, directed the differentiation of iPSCkit-EBs into Mesp1+/Isl1+/Nkx2.5+ cardiac mesoderm progenitors (p≤0.0001). Remarkably, the same signaling pathway subsequently directed EBs into the cKit+/Wnt1+/Pax3+/Mitf-H+/Isl1+/Nkx2.5+ CNC lineage (p≤0.0001), while suppressing the generation of WT1+/Tbx18+ epicardium (p<0.05). Stage-specific induction of Cre-recombination delineated that iPSCkit-derived CPCs encompass Mesp1–/cKit+/Nkx2.5+ CNC progenitors which contributed EGFP+ CNC derivatives, including Nkx2-5+ cardiomyocytes, to 60.7%±7.3% of spontaneously beating EBs (n=147 EBs; 12 preparations). Conclusions: Collectively, our data show that CPCkit are fully competent CNC-derived cardiomyogenic progenitors, whose differentiation to cardiomyocytes is minimized by a latent Noggin-mediated signaling pathway. Therefore exploiting CPCkit therapeutically, provides an important strategy for maximizing myocardial regeneration.

Abstract 19282: Transient Bone Morphogenic Protein Antagonism Directs Differentiation of Ipscs Into the Cardiac Neural Crest and Ckit+ Myocardial Progenitor Lineages

Circulation ◽  
2015 ◽  
Vol 132 (suppl_3) ◽  
Author(s):  
Konstantinos E Hatzistergos ◽  
Lauro M Takeuchi ◽  
Dieter Saur ◽  
Barbara Seidler ◽  
Susan M Dymecki ◽  
...  
Keyword(s):  
Neural Crest ◽  
Signaling Pathway ◽  
Progenitor Cell ◽  
Embryoid Bodies ◽  
Cardiac Neural Crest ◽  
Specific Induction ◽  
Cre Recombination ◽  
Bmp Antagonist ◽  
Cardiogenic Differentiation ◽  
Cardiac Mesoderm

Introduction: The signaling pathways that govern cKit+ cardiac progenitor cell (CPC) differentiation into cardiomyocytes (CMs) are unknown. Some studies suggest an essential role in cardiomyogenesis, others suggest a minimal CPC contribution. We studied if it is a non-permissive cardiac milieu that minimizes the generation of CMs from CPCs. Hypothesis: Transient BMP antagonism directs the generation of cardiomyocytes from CPCs. Methods: We lineage-traced CPCs using a novel dual-recombinase responsive indicator mice (cKitCreERT2;Wnt1::Flpe;RC::Fela) and iPSCs derived from cKitCreERT2;IRG (iPSCKit) mice. Results: Intersectional genetic fate-mapping of cKitCreERT2;Wnt1:: Flpe;RC::Fela embryos supported that cKit marks Wnt1-expressing cardiac neural crest (CNC) progenitors, emerging at ~E9.5 and contributing a limited number of cardiomyocytes (n=3). We lineage-traced CPCs during stage-specific cardiogenic differentiation of iPSCKit. Ascorbate treatment promoted differentiation of iPSCKit-derived embryoid bodies (EBs) into Nkx2.5+ myocardium, 45.5%±6.7% of which co-expressed the Cre-reporter EGFP (n=154 EBs; 12 preps), suggesting that CPCs encompass fully competent cardiomyogenic progenitors. Noggin (or Dorsomorphin), a BMP antagonist transiently expressed in the heart at E7.5-E8.5 but not during CNC invasion, directed the differentiation of iPSCkit-EBs into Mesp1+/Isl1+/Nkx2.5+ cardiac mesoderm progenitors (p≤0.0001). The same signaling pathway subsequently directed EBs into the cKit+/Wnt1+/Pax3+/Mitf-H+/Isl1+/Nkx2.5+ CNC lineage (p≤0.0001), while suppressing the generation of WT1+/Tbx18+ epicardium (p<0.05). Stage-specific induction of Cre-recombination delineated that iPSCkit-derived CPCs encompass Mesp1–/cKit+/Nkx2.5+ CNC progenitors, which contributed EGFP+ CNC derivatives, including Nkx2-5+ cardiomyocytes, to 60.7%±7.3% of spontaneously beating EBs (n=147 EBs; 12 preps). Conclusions: Our data show that CPCkit are fully competent CNC-derived cardiomyogenic progenitors, whose differentiation to cardiomyocytes is minimized by a latent Noggin-mediated signaling pathway. Therapeutically exploiting CPCkit, provides an important strategy for maximizing myocardial regeneration.

Abstract 441: Role of Nitric Oxide and S-nitrosylation in Cardiomyogenesis by iPSCs

2017 ◽  
Vol 121 (suppl_1) ◽  
Author(s):  
Alessandro Salerno ◽  
Konstantinos Hatzistergos ◽  
Raul Dulce ◽  
Amarylis Wanschel ◽  
Wayne Balkan ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Stem Cells ◽  
Signaling Pathways ◽  
Signaling Pathway ◽  
Es Cells ◽  
Embryoid Bodies ◽  
Negative Regulator ◽  
Cell Based Therapy ◽  
Cardiac Phenotype ◽  
Cardiac Mesoderm

Introduction: The mechanism by which signaling pathways, such as Wnt and BMP interact and modulate each other’s function is crucial to our understanding of cardiomyogenesis and cardiomyocyte proliferation. Nitric oxide (NO) is a signaling molecule that can trigger cardiac differentiation of stem cells, suggesting a cardiogenic function of NO synthase(s) (NOS). Hypothesis: NO modulates transcription factor function during pluripotency and differentiation toward a cardiac phenotype. Methods: Induced pluripotent stem cells (iPSCs) were derived from fibroblasts from wildtype mice and mice lacking S-nitrosoglutathione reductase (GSNOR -/- ), a denitrosylase that regulates protein S-nitrosylation. iPSCs were differentiated into functional cardiomyocytes from embryoid bodies (EBs) via the hanging-drop method. Results: During differentiation into cardiomyocytes, GSNOR -/- iPSC-derived cardiomyocytes exhibited reduced expression of mesoderm induction-related ( Brachyury ), cardiac mesoderm (Kdr , Isl-1 ) and cardiac progenitor genes ( Nkx2.5 , GATA4 ). Axin-1, an inducer of apoptosis and negative regulator of the Wnt signaling pathway and MAPK pathways, specifically p38, were increased on EB-Day (D)4. In contrast, SMAD1/5/8, members of the BMP canonical signaling pathway, were reduced beginning on EB-D8. Increased p38 is associated with reduced GATA4 expression and differentiation of human ES cells into cardiomyocytes. Decreased SMAD1/5/8 is likely at least in part responsible for the reduced expression of Nkx2.5. Conclusions: Our findings support that the absence of GSNOR modulates Wnt/β-catenin and BMP signaling pathways during cardiogenesis, resulting in reduced expression of mesoderm, cardiac mesoderm and cardiac progenitor genes. These findings are expected to have important implications for regenerative medicine and can provide new targets for iPS cell-based therapy.

Abstract 323: Loss of Gravity Impairs Cardiac Neural Crest Cell Lineage Development and Function

2016 ◽  
Vol 119 (suppl_1) ◽  
Author(s):  
Konstantinos E Hatzistergos ◽  
Krystalenia Valasaki ◽  
Zhijie Jiang ◽  
Lauro M Takeuchi ◽  
Wayne Balkan ◽  
...  
Keyword(s):  
Neural Crest ◽  
Molecular Mechanisms ◽  
Cell Lineage ◽  
Neural Crest Cell ◽  
Lineage Tracing ◽  
Embryoid Bodies ◽  
Egfp Expression ◽  
Cardiac Neural Crest ◽  
Induced Pluripotent ◽  
And Function

Introduction: A multitude of structural, haemodynamic and electromechanical cardiovascular disorders have been observed in humans following space-travel. These abnormalities are thought to emerge from transient alterations in autonomic nervous system (ANS). However, since the ANS is cardiac neural crest (CNC)-derived, whether microgravity-induced cardiomyopathies reflect CNC dysfunction, is unknown. Hypothesis: Impairment of CNCs underlies microgravity-induced cardiomyopathies. Methods: Myocardial explants from adult cKit CreERT2/+ ;IRG mice (n=5/group), as well as cKit CreERT2/+ ;IRG- derived (iPSC Kit-Cre ; n=6/group) and Wnt1-Cre;tdTomato -derived (iPSC Wnt1-Cre ; n=18/group) induced pluripotent stem cells, were cultured under static (SC) or simulated microgravity conditions (rotary cell-culture system; RCCS). Results: CNC lineage-tracing in cardiac explants illustrated that, compared to SC, RCCS abolished the pool of cKit + CNCs in adult hearts, indicated by quantitation of cKit CreERT2 - mediated EGFP expression ( p <0.05). Cardiogenesis modeling experiments with iPSC Kit-Cre yielded fewer beating EBs ( p =0.0005), and ~10-fold reduction in EGFP + cardiomyocytes ( p =0.01), in RCCS vs . SC. Microarray analyses suggested that RCCS-mediated alterations in BMP and Wnt/β-catenin pathways, downregulated ANS and CNC-related gene programs, and enhanced vasculogenic differentiation without affecting the expression of cardiac mesoderm-related genes. Differences were verified by quantitative PCR. Modeling CNC development in iPSC Wnt1-Cre further confirmed an RCCS-mediated dramatic impairment in development and function of CNCs, indicated by quantitation of tdTomato expression in day-10 and day-21 beating embryoid bodies ( p <0.0001). Intriguingly, the effect of RCCS in CNCs could be only partially rescued upon transfer to SC. Conclusions: Together these data indicate that microgravity negatively regulates the development and function of CNCs, thus partly explaining the cellular and molecular mechanisms of microgravity-induced cardiomyopathies. Moreover, these findings are expected to have important implications in space exploration, since they suggest an essential role for gravity in vertebrate development.

scholarly journals Effects of the size of lesions of the cardiac neural crest at various embryonic ages on incidence and type of cardiac defects.

Circulation ◽  
1986 ◽  
Vol 73 (2) ◽  
pp. 360-364 ◽  
Author(s):  
W T Besson ◽  
M L Kirby ◽  
L H Van Mierop ◽  
J R Teabeaut
Keyword(s):  
Neural Crest ◽  
Cardiac Defects ◽  
Cardiac Neural Crest

scholarly journals Foxc2 is required for proper cardiac neural crest cell migration, outflow tract septation, and ventricle expansion

2018 ◽  
Vol 247 (12) ◽  
pp. 1286-1296 ◽  
Author(s):  
Kimberly E. Inman ◽  
Carlo Donato Caiaffa ◽  
Kristin R. Melton ◽  
Lisa L. Sandell ◽  
Annita Achilleos ◽  
...  
Keyword(s):  
Cell Migration ◽  
Neural Crest ◽  
Neural Crest Cell ◽  
Outflow Tract ◽  
Cardiac Neural Crest ◽  
Neural Crest Cell Migration ◽  
Crest Cell

scholarly journals Twist1 performs distinct spatio‐temporal functions in developing cardiac neural crest cells

2011 ◽  
Vol 25 (S1) ◽  
Author(s):  
Joshua Wayne Vincentz ◽  
Ralston Barnes ◽  
Beth Firulli ◽  
Douglas Spicer ◽  
Anthony Firulli
Keyword(s):  
Neural Crest ◽  
Neural Crest Cells ◽  
Cardiac Neural Crest ◽  
Spatio Temporal

scholarly journals Notch Signaling Commits Mesoderm to the Cardiac Lineage

2020 ◽  
Author(s):  
Evan S. Bardot ◽  
Bharati Jadhav ◽  
Nadeera Wickramasinghe ◽  
Amélie Rezza ◽  
Michael Rendl ◽  
...  
Keyword(s):  
Notch Signaling ◽  
Heart Development ◽  
Cardiomyocyte Differentiation ◽  
Mouse Development ◽  
Mesoderm Specification ◽  
Cardiac Lineage ◽  
Underlying Mechanisms ◽  
Early Mouse ◽  
Differentiation Efficiency ◽  
Cardiac Mesoderm

AbstractDuring development multiple progenitor populations contribute to the formation of the four-chambered heart and its diverse lineages. However, the underlying mechanisms that result in the specification of these progenitor populations are not yet fully understood. We have previously identified a population of cells that gives rise selectively to the heart ventricles but not the atria. Here, we have used this knowledge to transcriptionally profile subsets of cardiac mesoderm from the mouse embryo and have identified an enrichment for Notch signaling components in ventricular progenitors. Using directed differentiation of human pluripotent stem cells, we next investigated the role of Notch in cardiac mesoderm specification in a temporally controlled manner. We show that transient Notch induction in mesoderm increases cardiomyocyte differentiation efficiency, while maintaining cardiomyocytes in an immature state. Finally, our data suggest that Notch interacts with WNT to enhance commitment to the cardiac lineage. Overall, our findings support the notion that key signaling events during early heart development are critical for proper lineage specification and provide evidence for early roles of Notch and WNT during mouse and human heart development.Summary statementEarly fate decisions are dictated by the embryonic signaling environment. We show that Notch signaling is active during early mouse development and that activating Notch in human cardiac mesoderm enhances cardiomyocyte differentiation efficiency.

scholarly journals Epigenetic Regulation of Cardiac Neural Crest Cells

2021 ◽  
Vol 9 ◽  
Author(s):  
Shun Yan ◽  
Jin Lu ◽  
Kai Jiao
Keyword(s):  
Neural Crest ◽  
Epigenetic Regulation ◽  
Heart Development ◽  
Congenital Heart ◽  
Heart Diseases ◽  
Heart Defects ◽  
Neural Crest Cells ◽  
Abnormal Development ◽  
Cardiac Neural Crest ◽  
Epigenetic Regulators

The cardiac neural crest cells (cNCCs) is a transient, migratory cell population that contribute to the formation of major arteries and the septa and valves of the heart. Abnormal development of cNCCs leads to a spectrum of congenital heart defects that mainly affect the outflow region of the hearts. Signaling molecules and transcription factors are the best studied regulatory events controlling cNCC development. In recent years, however, accumulated evidence supports that epigenetic regulation also plays an important role in cNCC development. Here, we summarize the functions of epigenetic regulators during cNCC development as well as cNCC related cardiovascular defects. These factors include ATP-dependent chromatin remodeling factors, histone modifiers and DNA methylation modulators. In many cases, mutations in the genes encoding these factors are known to cause inborn heart diseases. A better understanding of epigenetic regulators, their activities and their roles during heart development will ultimately contribute to the development of new clinical applications for patients with congenital heart disease.

Sign in / Sign up

Export Citation Format

Share Document