scholarly journals Sox17 expression in endocardium precursor cells regulates heart development in mice

2019 ◽  
Author(s):  
Rie Saba ◽  
Keiko Kitajima ◽  
Lucille Rainbow ◽  
Sylvia Engert ◽  
Mami Uemura ◽  
...  
Keyword(s):  
Gene Expression ◽  
Progenitor Cells ◽  
Heart Development ◽  
Molecular Mechanisms ◽  
Homeobox Gene ◽  
Cardiac Progenitor Cells ◽  
Cellular Behavior ◽  
Hmg Box ◽  
And Behavior ◽  
Morphology Of The Heart

AbstractThe endocardium is the endothelial component of the vertebrate heart and plays a key role in heart development. Cardiac progenitor cells (CPCs) that express the homeobox gene Nkx2-5 give rise to the endocardium. Where, when, and how the endocardium segregates during embryogenesis have remained largely unknown, however. We now show that Nkx2-5+ CPCs that express the Sry-type HMG box gene Sox17 specifically differentiate into the endocardium in mouse embryos. Approximately 20% to 30% of Nkx2-5+CPCs transiently express Sox17 from embryonic day (E) 7.5 to E8.5.Although Sox17 is not essential or sufficient for endocardium fate, it can bias the fate of CPCs toward the endocardium. On the other hand, Sox17 expression in the endocardium is required for heart development. Deletion of Sox17 specifically in the mesoderm markedly impaired endocardium development with regard to cell proliferation and behavior. The proliferation of cardiomyocytes, ventricular trabeculation, and myocardium thickening were also impaired in a non–cell-autonomous manner in the Sox17 mutant, resulting in anomalous morphology of the heart, likely as a consequence of down-regulation of NOTCH signaling. Changes in gene expression profile in both the endocardium and myocardium preceded the reduction in NOTCH-related gene expression in the mutant embryos, suggesting that Sox17 expression in the endocardium regulates an unknown signal required for nurturing of the myocardium. Our results thus provide insight into differentiation of the endocardium and its role in heart development.SignificanceThe endocardium is vital for vertebrate heart development; however, the molecular mechanisms regulating fate determination and differentiation remain largely unknown. Here, we show that a part of the earliest cardiac progenitor cells (CPCs) transiently and exclusively express Sry-type HMG box gene Sox17 in the mouse embryo. Sox17-expressing CPCs specifically differentiate to the endocardium. Sox17 biases the fate of CPCs toward the endocardium, and regulates proliferation and cellular behavior cell autonomously. Conversely, Sox17 in the endocardium regulates the myocardium non-cell autonomously. Notably, Sox17 is required for the ventricular trabeculation via the NOTCH signal that is not directly induced but maintained by Sox17. This study, thus, sheds light on endocardium development.

scholarly journals Differential rescue of visceral and cardiac defects inDrosophilaby vertebratetinman-related genes

1998 ◽  
Vol 95 (16) ◽  
pp. 9366-9371 ◽  
Author(s):  
Maijon Park ◽  
Carol Lewis ◽  
David Turbay ◽  
Amy Chung ◽  
Jau-Nian Chen ◽  
...  
Keyword(s):  
Gene Expression ◽  
Heart Development ◽  
Molecular Mechanisms ◽  
Marker Gene ◽  
Homeobox Gene ◽  
Specific Gene ◽  
Specific Marker ◽  
Cardiac Progenitors ◽  
Mesoderm Development ◽  
Visceral Mesoderm

tinman, a mesodermal NK2-type homeobox gene, is absolutely required for the subdivision of the earlyDrosophilamesoderm and for the formation of the heart as well as the visceral muscle primordia. Several vertebrate relatives oftinman, many of which are predominately expressed in the very early cardiac progenitors (and pharyngeal endoderm), also seem to promote heart development. Here, we show that most of these vertebratetinman-related genes can readily substitute forDrosophila tinmanfunction in promoting visceral mesoderm-specific marker gene expression, but much less in promoting cardiac-specific gene expression indicative of heart development. In addition, another mesodermal NK2-type gene fromDrosophila,bagpipe, which is normally only needed for visceral mesoderm but not heart development, cannot substitute fortinmanat all. These data indicate that the functional equivalence of thetinman-related subclass of NK2-type genes (in activating markers of visceral mesoderm development inDrosophila) is specific to this subclass and distinct from other homeobox genes. Despite the apparent overall conservation of heart development between vertebrates and invertebrates, the differential rescue of visceral mesoderm versus heart development suggests that some of the molecular mechanisms of organ formation may have diverged during evolution.

Abstract 247: Scaffold-stiffness Dependent Notch1 Activation Regulates Cardiogenic Gene Expression In Cardiac Progenitor Cells

2013 ◽  
Vol 113 (suppl_1) ◽  
Author(s):  
Archana V Boopathy ◽  
Pao L Che ◽  
Yoshie Narui ◽  
Khalid Salaita ◽  
Michael E Davis
Keyword(s):  
Gene Expression ◽  
Smooth Muscle ◽  
Cardiac Function ◽  
Progenitor Cells ◽  
Adult Stem Cells ◽  
Cardiac Progenitor Cells ◽  
Critical Pathway ◽  
Cardiac Gene Expression ◽  
Self Assembling ◽  
Cardiac Gene

Rationale: Cardiac progenitor cells (CPCs) are multipotent, self-renewing cells that can regenerate the myocardium and improve cardiac function in animal models of myocardial infarction (MI). However, limited survival of stem/progenitor cells inhibits cardiac regeneration. Force dependent Notch activation promotes cardiac development and cardiac gene expression in many adult stem cells. As dysregulation of Notch signaling leads to embryonic lethal cardiovascular defects, activating this critical pathway during cell transplantation could improve efficacy of stem cell therapy. Objective: Investigate i) whether self-assembling peptide scaffolds can be used to activate Notch1 signaling in CPCs to promote cardiogenic differentiation and ii) the effect of scaffold stiffness on Notch1 activation and differentiation. Methods: Rat CPCs (c-kit + ) were cultured for 48h in 3D self-assembling scaffolds of varying stiffness (1% low, 2% high): empty scaffolds (RADA), scaffolds modified with peptide mimicking Notch1 ligand, Jagged1 (RJAG), or scaffolds modified with a scrambled peptide (RSCR) and cardiogenic gene expression measured by qRT-PCR. CHO cells expressing Notch1 responsive YFP were also cultured in the above scaffolds for 48h and YFP expression was determined. Results are mean ± SEM with p<0.05 considered significant by one or two-way ANOVA with appropriate post test. Results: In the Notch1 reporter cells, Notch1 activation increased significantly in presence of RJAG (p<0.01) and on increasing scaffold stiffness (p<0.01,n=6) indicating scaffold stiffness-dependent Notch1 activation. Culture of CPCs in RJAG containing 1% scaffolds (low stiffness) significantly increased early endothelial and smooth muscle but not cardiac gene expression while in 2% scaffolds (high stiffness) significantly increased only cardiac and not endothelial or smooth muscle gene expression (p<0.05, n≥4). Conclusions: Taken together, these data show that i) Notch1 activation in 3D is dependent on ligand density and scaffold stiffness and ii) stiffness dependent Notch1 activation differentially regulates cardiogenic gene expression in CPCs. Therefore, delivery of CPCs in JAG containing scaffolds could be used to improve cardiac function following MI.

scholarly journals Transcription factor-induced activation of cardiac gene expression in human c-kit+ cardiac progenitor cells

PLoS ONE ◽  
2017 ◽  
Vol 12 (3) ◽  
pp. e0174242 ◽  
Author(s):  
Tareq Al-Maqtari ◽  
Kyung U. Hong ◽  
Bathri N. Vajravelu ◽  
Afsoon Moktar ◽  
Pengxiao Cao ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transcription Factor ◽  
Progenitor Cells ◽  
Cardiac Progenitor Cells ◽  
Cardiac Gene Expression ◽  
Cardiac Gene

Abstract 29: Porcine Decellularized Heart Tissue Enhance the Expression of Contractile Proteins in Human Cardiomyocytes and Differentiated Cardiac Progenitor Cells

2016 ◽  
Vol 119 (suppl_1) ◽  
Author(s):  
Anne-Cecile Huby ◽  
Farideh Beigi ◽  
Qian Xiang ◽  
Andrea Gobin ◽  
Doris Taylor
Keyword(s):  
Gene Expression ◽  
Progenitor Cells ◽  
Ips Cells ◽  
Contractile Proteins ◽  
Cardiac Cells ◽  
Contractile Protein ◽  
Cell Phenotype ◽  
Cell Physiology ◽  
Cardiac Progenitor Cells ◽  
Structural Support

Tissue engineering is an evolving tool for repair of cardiovascular damage, in particular after myocardial infarction. Currently, most studies are focused on providing gel or thin synthetic matrices to provide structural support for injured valves, or ventricle. But little is known about how cardiac cells (eg, endothelial cells [ECs], cardiomyocytes [CMs] and cardiac progenitor cells [CPCs]) are modified by an exposure to a scaffold much less how a xenogeneic scaffold may affect cell physiology or function. Our study was designed to examine the effect of rat or pig collagen matrix and decellularized left ventricle extracellular matrix (dECM) tissues on the phenotype of human cardiac cells. Human ECs, CMs and CPCs derived from IPs cells (Cellular Dynamics International) were cultured singly or together on rat or pig collagen matrices or on dECM. Protein and gene expression was investigated by immunohistochemistry, histology and real time PCR. All 3 cell types survived on both types of support when delivered singly or together. Endothelial cell PECAM-1 expression was enhanced on decellularized tissues particularly on rat matrix. Cardiomyocyte contractile protein expression did not differ with support type, whereas CPC contractile protein gene expression increased on both decellularized tissues more so on pig matrix. Additionally, when the cells were co-cultured, single cell type gene expression was modified. Our study showed that human cardiac cells are influenced by the structural support on which they are cultivated. Cardiac ECM enhanced the expression of contractile proteins and endothelial markers but matrix species had minimal influence on this effect. Furthermore, cell co-culture altered gene expression suggesting cell communication is intact on the matrix. This study shows that cell phenotype and behavior is impacted by the tissue matrix to a greater degree than occurs with a simple one protein support and suggests complex decellularized matrix may be superior to successfully engineer cardiac constructs with intact physiology. Additionally, the successful differentiation of IPs cells into cardiomyocytes on decellularized matrix opens avenue to build tissue engineer constructs from patients with heart disease as a screening tool.

Abstract 212: Hydrogen Peroxide Acutely Stimulates Cardiogenic Gene Expression in Cardiac Progenitor Cells

2012 ◽  
Vol 111 (suppl_1) ◽  
Author(s):  
Karl D Pendergrass ◽  
Michael E Davis
Keyword(s):  
Gene Expression ◽  
Heart Failure ◽  
Hydrogen Peroxide ◽  
Smooth Muscle ◽  
Progenitor Cells ◽  
Matched Control ◽  
Cardiac Progenitor Cells ◽  
H2o2 Treatment ◽  
Apoptotic Effect ◽  
H2o2 Pretreatment

Following acute myocardial infarction, billions of myocytes are lost to cell death. The damage is regional and lost cells are replaced with a collagen scar. One potential therapy to delay or prevent progression into heart failure is regeneration of the damaged myocardium through cell therapy with cardiac progenitor cells (CPC). Reactive oxygen species, specifically hydrogen peroxide, elicit varying responses from different stem/progenitor cells. In the present studies, we sought to determine the effect of acute H2O2 treatment on CPC survival and differentiation. CPCs were isolated and cultured with leukemia inhibitory factor (LIF) to retain their stem-like qualities. CPCs were allowed to differentiate in the absence of LIF for up to 5 days + H2O2. H2O2 (100μM) significantly increased expression of the smooth muscle marker, alpha smooth muscle actin (αSM) by Day 2 as compared to time-matched controls (Ctl: 1.8+1.3 vs 100 μM: 8.5+1.1; p<0.001; N=3). We also observed a trend for an increase in smooth muscle 22 alpha (SM 22α) gene expression by Day 2. Interestingly, by Day 5 the stimulatory effect of 100 μM H2O2 treatment on α SM and SM 22α was reversed and significantly decreased compared to Day 2 (D5: α SM: 0.43+0.05, SM 22α: 0.41+0.2 vs. D2: SM 22α: 10.58+2.3; p<0.01; N=3-4). Evaluation of the endothelial marker VEGFR-2 (Flk-1) showed a trend for an increase in gene expression by Day 2 following 100 μM H2O2 treatment compared to the time-matched control. We also observed an anti-apoptotic effect on CPCs following serum removal, in which 2 days of 100μM H2O2 pretreatment lead to approximately a 55% decrease in cell death compared to untreated CPCs (Ctl: 19.1+2.4 vs 100 μM: 10.5+1.7; p<0.05; N=3-4). The protective effect of the H2O2 pretreatment could be attributed to an increase in anti-oxidative enzymatic capacity in CPCs. There was a trend for an increase in catalase gene expression. In conclusion, our results showed that acute H2O2 preconditioning exerted a stimulatory effect on smooth muscle gene expression and an anti-apoptotic effect compared to time-matched Control CPCs. Furthermore, acute H2O2 preconditioning may aid in directing CPC differentiation towards a vascular phenotype and angiogenesis in the infarcted myocardium, which may prevent or delay heart failure.

Abstract 21: Stiffness-Dependent Notch1 Activation Regulates Cardiogenic Differentiation of Cardiac Progenitor Cells

2012 ◽  
Vol 111 (suppl_1) ◽  
Author(s):  
Archana V Boopathy ◽  
Khalid Salaita ◽  
Michael E Davis
Keyword(s):  
Gene Expression ◽  
Amino Acid ◽  
Cardiac Function ◽  
Notch Signaling ◽  
Progenitor Cells ◽  
Cardiac Progenitor Cells ◽  
Critical Pathway ◽  
3D Scaffolds ◽  
Self Assembling ◽  
Cardiogenic Differentiation

Cardiac progenitor cells (CPCs) are multipotent, self-renewing cells that can regenerate the myocardium and improve cardiac function in animal models of MI by cardiogenic differentiation. However, limited survival of stem/progenitor cells, myocardial scarring and fibrosis inhibit cardiac regeneration. Notch signaling promotes early cardiac development, cardiomyocyte survival and cardiac gene expression in circulating endothelial progenitor cells, mesenchymal stem cells and CPCs. As misregulation of Notch signaling during development is lethal due to cardiovascular defects, activating this critical pathway during cell transplantation could improve the efficacy of stem cell therapy. We investigated whether self-assembling peptide nanofiber hydrogels can be used to activate Notch1 signaling. The 16 amino acid self-assembling scaffold (RAD) was modified with a 20 amino acid peptide mimicking the active site of Notch1 ligand, Jagged1 (RJAG) or with the corresponding scrambled peptide (RSCR). To determine whether scaffold stiffness regulates Notch1 activation, CHO cells with Notch1 responsive YFP expression were cultured in scaffolds of 1-3% w/v in presence of RSCR or RJAG at a 1:10 ligand: scaffold ratio in 3D. Presence of the RJAG peptide (p<0.01) and % concentration of the scaffold (p<0.01) increased Notch1 activation significantly (n=5) indicating that RJAG mediated Notch1 activation in 3D is scaffold stiffness-dependent. Therefore, CPCs were cultured within 3D scaffolds (1-3% w/v; empty, scaffold +RJAG or RSCR) and cardiogenic gene expression was determined by qPCR. An increase in expression of early endothelial (Flk1, Flt1, vWF) and smooth muscle (sm22α, sm αactin) genes was observed in CPCs cultured in 3D scaffolds containing RJAG but not when cultured in 2D. These data show that Notch1 activation is dependent on ligand density and scaffold stiffness. Delivery of CPCs in JAG1 containing self-assembling scaffolds could be used to enhance therapeutic angiogenesis and improve cardiac function following myocardial infarction.

From the Bone Marrow to the Thymic Niche

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 5123-5123
Author(s):  
Sandrine Susini ◽  
Séverine Mouraud ◽  
Elodie Elkaim ◽  
Julien Roullier ◽  
Sonia Luce ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bone Marrow ◽  
Epithelial Cells ◽  
Progenitor Cells ◽  
Chemokine Receptors ◽  
Molecular Mechanisms ◽  
Conflicts Of Interest ◽  
Adult Life ◽  
Thymic Epithelial Cells

Abstract To generate T cells throughout adult life, the thymus must import hematopoietic progenitor cells from the bone marrow via the blood. The cellular and molecular mechanisms governing the circulation of thymus-seeding progenitor cells are well characterized in mice but not in humans. The aim of the present study was to characterize the molecular mechanisms and cellular components involved in thymus colonization by lymphoid progenitors (CD34+/CD10+/CD7-/CD24-) and the early steps of thymopoiesis under physiological conditions in humans. Our results demonstrate that circulating lymphoid progenitor cells express CCR9 and CXCR4 chemokine receptors, VLA-4, VLA-5 and VLA-6 integrins and PSGL-1 and CD44 adhesion molecules. We used in vitro migration and adhesion assays to validate the functional status of these markers. As in the mouse, human circulating progenitor cells enter the thymus at the corticomedullary junction (CMJ). Once in the thymus, crosstalk with thymic epithelial cells causes the circulating progenitors to commit to the T-cell differentiation pathway. In order to characterize thymic niches and interactions between circulating progenitors and the thymic stroma, we undertook a chemokine/chemokine-receptor-focused gene expression analysis of sorted lymphoid progenitor cells and CMJ epithelial cells (based on the expression of EpCAM and Delta-like-4). We observed an unexpected gene expression profile for chemokines and chemokine regulators in thymus-seeding CD34+/CD10+/CD7-/CD24- cells and epithelial cells at the CMJ. The present results should help us to highlight candidate genes involved in the early steps of human thymopoiesis. Disclosures No relevant conflicts of interest to declare.

scholarly journals The effect of neurogenin3 deficiency on pancreatic gene expression in embryonic mice

2006 ◽  
Vol 37 (2) ◽  
pp. 301-316 ◽  
Author(s):  
Andreas Petri ◽  
Jonas Ahnfelt-Rønne ◽  
Klaus Stensgaard Frederiksen ◽  
David George Edwards ◽  
Dennis Madsen ◽  
...  
Keyword(s):  
Gene Expression ◽  
Microarray Analysis ◽  
Molecular Mechanisms ◽  
Homeobox Gene ◽  
Specific Gene ◽  
Wild Type ◽  
Specific Gene Expression ◽  
And Function ◽  
Deficient Mice

To understand the molecular mechanisms regulating pancreatic endocrine development and function, pancreatic gene expression was compared between Ngn3-deficient mice and littermate controls on embryonic days 13 and 15. Microarray analysis identified 504 genes with significant differences in expression. Fifty-two of these showed at least twofold reduction in Ngn3 knockouts compared to controls. Many of them were previously described to be involved in endocrine development and function. Among the genes not previously characterized were Rhomboid veinlet-like 4, genes involved in tetrahydrobiopterin biosynthesis and the Iroquois-type homeobox gene Irx1, the latter was selected for further investigation. In situ hybridisation demonstrated that two Iroquois genes, Irx1 and Irx2, were expressed in pancreatic endoderm of wild-type, but not Ngn3 mutant embryos. Furthermore, ectopic Ngn3 induced prominent Irx2 expression in chicken endoderm. Co-labelling established that Irx1 and Irx2 mRNA is located to glucagon-, but not insulin- or somatostatin-producing cells in mice and chicken. These data suggest that Irx1 and Irx2 serve an evolutionary conserved role in the regulation of α-cell-specific gene expression.

scholarly journals Analysis of the SWI/SNF chromatin-remodeling complex during early heart development and BAF250a repression cardiac gene transcription during P19 cell differentiation

2013 ◽  
Vol 42 (5) ◽  
pp. 2958-2975 ◽  
Author(s):  
Ajeet Pratap Singh ◽  
Trevor K. Archer
Keyword(s):  
Gene Expression ◽  
Cell Differentiation ◽  
Chromatin Remodeling ◽  
Heart Development ◽  
Molecular Mechanisms ◽  
Gene Expression Regulation ◽  
Affinity Purification ◽  
Specific Gene ◽  
Nucleosome Remodeling ◽  
Cardiac Gene

Abstract The regulatory networks of differentiation programs and the molecular mechanisms of lineage-specific gene regulation in mammalian embryos remain only partially defined. We document differential expression and temporal switching of BRG1-associated factor (BAF) subunits, core pluripotency factors and cardiac-specific genes during post-implantation development and subsequent early organogenesis. Using affinity purification of BRG1 ATPase coupled to mass spectrometry, we characterized the cardiac-enriched remodeling complexes present in E8.5 mouse embryos. The relative abundance and combinatorial assembly of the BAF subunits provides functional specificity to Switch/Sucrose NonFermentable (SWI/SNF) complexes resulting in a unique gene expression profile in the developing heart. Remarkably, the specific depletion of the BAF250a subunit demonstrated differential effects on cardiac-specific gene expression and resulted in arrhythmic contracting cardiomyocytes in vitro. Indeed, the BAF250a physically interacts and functionally cooperates with Nucleosome Remodeling and Histone Deacetylase (NURD) complex subunits to repressively regulate chromatin structure of the cardiac genes by switching open and poised chromatin marks associated with active and repressed gene expression. Finally, BAF250a expression modulates BRG1 occupancy at the loci of cardiac genes regulatory regions in P19 cell differentiation. These findings reveal specialized and novel cardiac-enriched SWI/SNF chromatin-remodeling complexes, which are required for heart formation and critical for cardiac gene expression regulation at the early stages of heart development.

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