Erratum to "Modulation of Human Cardiac Progenitors via Hypoxia-ERK Circuit Improves their Functional Bioactivities" [Biomol. Ther. 21 (2013) 196-203]

The circulatory system is the first functional organ system that develops during mammalian life. Accumulating evidences suggest that cardiac and endocardial cells can arise from a single common progenitor cell during mammalian cardiogenesis. Notably, these early cardiac progenitors express multiple hematopoietic transcription factors, consistent with previous reports. Indeed, a close relationship among cardiac, endocardial and hematopoietic lineages has been suggested in fly, zebrafish, and embryonic stem cell in vitro differentiation models. However, it is unclear when, where and how this hematopoietic gene program is in operation during in vivo mammalian cardiogenesis. Hematopoietic colony assay suggests that mouse heart explants generate myeloids and erythroids in the absence of circulation, suggesting that the heart tube is a de novo site for the definitive hematopoiesis. Lineage tracing revealed that putative cardiac-derived Nkx2-5+/Isl1+ endocardial cells give rise to CD41+ hematopoietic progenitors that contribute to definitive hematopoiesis in vivo and ex vivo during embryogenesis earlier than in the AGM region. Furthermore, Nkx2-5 and Isl1 are both required for the hemogenic activity of the endocardium. Together, identification of Nkx2-5/Isl1-dependent hemogenic endocardial cells (1) adds hematopoietic component in the cardiogenesis lineage tree, (2) changes the long-held dogma that AGM is the only major source of definitive hematopoiesis in the embryo proper, and (3) represents phylogenetically conserved fundamental mechanism of cardio-vasculo-hematopoietic differentiation pathway during the development of circulatory system.

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Abstract 17802: Ips Cells -derived Cardiac Progenitors Induce Dramatic Cardiac Regeneration and Improve Function by Cxcr4 Signaling Pathway in Infarcted Myocardium

Circulation ◽

10.1161/circ.130.suppl_2.17802 ◽

2014 ◽

Vol 130 (suppl_2) ◽

Author(s):

Zeshan Pasha ◽

Romana Saeed ◽

Muhammad Ashraf

Keyword(s):

Stem Cells ◽

Signaling Pathway ◽

Small Molecule ◽

In Vivo Studies ◽

Immunofluorescent Staining ◽

Concomitant Treatment ◽

Post Transplantation ◽

Cardiac Progenitors ◽

Cxcr4 Signaling ◽

Infarcted Myocardium

Background: The participation of endogenous cardiac stem/progenitor cells is limited in restoring cardiac structure and function in the ischemic myocardium which is further aggravated by poor survival and propagation of transplanted stem cells of different origin in the infarcted heart. The goal of this study was to explore the survival and engraftability of newly discovered induced pluripotent stem cells (IPS) in the myocardium following infarction (MI). Methods and Results: Integration free iPS were generated from myoblasts and characterized. Cardiac progenitors (CPs) were created by treatment with a small molecule. CPs proliferation was assessed by BrdU labeling; Differentiation by both RT-PCR and immunofluorescent staining for cardiac markers Nkx2.5, actinin, and -MHC. Gene expression profiling was performed using Affymetrix array. In vivo studies were carried out by injecting CPs or nontreated IPS (3x105), into mouse model of permanent LAD. Echocardiography, histological parameters, TUNEL assay and capillary vessel density were measured 6 weeks post transplantation. Treatment of IPS with a small molecule upregulated Nkx2.5 and maintained up to 4 weeks (p<0.01 vs nontreated IPS). Expression of actinin and -MHC was also detectable at 3 weeks. Increased proliferative activity (p<0.01) evaluated by cell proliferation and Brdu assay was observed. Significant CPs survival and reduced apoptosis were noticed in the small molecule treated iPSC compared to nontreated and/or saline group. Enhanced ejection fraction and fractional shortening (p< 0.05) was observed 6 weeks post transplantation. Interestingly there was 2-3 fold upregulation of chemokines including CCL7, CXCR2, CXCR5. miR Microarray analysis showed upregulation of cardiac specific mir-133,762. Western blot analysis showed increased phospho-Akt levels as compared to nontreated IPSC (p<0.01). Survival and differentiation properties of CPs were abolished by concomitant treatment of IPS with CXR4 blocker (p<0.05). Conclusion: This study provides a novel strategy for generating CPs and their enhanced survival, engraftment and differentiation with the treatment of cardiogenic small molecule post transplantation in the infarcted myocardium through CXCR4 signaling pathway.

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Abstract 257: Clonal Analysis Of Multipotent Cardiovascular Progenitors That Generate Cardiomyocytes During Development

Circulation Research ◽

10.1161/res.115.suppl_1.257 ◽

2014 ◽

Vol 115 (suppl_1) ◽

Author(s):

Konstantina Ioanna Sereti ◽

Paniz Kamran Rashani ◽

Peng Zhao ◽

Reza Ardehali

Keyword(s):

Single Cell ◽

Heart Development ◽

Cardiac Development ◽

Clonal Analysis ◽

Cardiac Cells ◽

Single Cell Level ◽

Reporter System ◽

Cell Level ◽

Cardiac Progenitors

It has been proposed that cardiac development in lower vertebrates is driven by the proliferation of cardiomyocytes. Similarly, cycling myocytes have been suggested to direct cardiac regeneration in neonatal mice after injury. Although, the role of cardiomyocyte proliferation in cardiac tissue generation during development has been well documented, the extent of this contribution as well as the role of other cell types, such as progenitor cells, still remains controversial. Here we used a novel stochastic four-color Cre-dependent reporter system (Rainbow) that allows labeling at a single cell level and retrospective analysis of the progeny. Cardiac progenitors expressing Mesp1 or Nkx2.5 were shown to be a source of cardiomyocytes during embryonic development while the onset of αMHC expression marked the developmental stage where the capacity of cardiac cells to proliferate diminishes significantly. Through direct clonal analysis we provide strong evidence supporting that cardiac progenitors, as opposed to mature cardiomyocytes, are the main source of cardiomyocytes during cardiac development. Moreover, we have identified quadri-, tri-, bi, and uni-potent progenitors that at a single cell level can generate cardiomyocytes, fibroblasts, endothelial and smooth muscle cells. Although existing cardiomyocytes undergo limited proliferation, our data indicates that it is mainly the progenitors that contribute to heart development. Furthermore, we show that the limited proliferation capacity of cardiomyocytes observed during normal development was enhanced following neonatal cardiac injury allowing almost complete regeneration of the scared tissue. However, this ability was largely absent in adult injured hearts. Detailed characterization of dividing cardiomyocytes and proliferating progenitors would greatly benefit the development of novel therapeutic options for cardiovascular diseases.

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Abstract 304: Efficient Generation of Cardiac Purkinje Fiber-like Cells From ESCs by Activating cAMP Signaling

Circulation Research ◽

10.1161/res.117.suppl_1.304 ◽

2015 ◽

Vol 117 (suppl_1) ◽

Author(s):

Su-Yi Tsai ◽

Karen Maass ◽

Jia Lu ◽

Glenn I Fishman ◽

Shuibing Chen ◽

...

Keyword(s):

Embryonic Stem ◽

Cardiac Cells ◽

Cardiac Conduction ◽

Camp Signaling ◽

Purkinje Fiber ◽

Mechanistic Studies ◽

Cardiac Progenitors ◽

Gene Profiles ◽

Novel Strategy ◽

Regenerative Therapies

Dysfunction of the cardiac conduction system (CCS) significantly impacts pathogenesis of arrhythmia, a major cause of morbidity and mortality. Strategies to derive cardiac conduction cells including Purkinje fiber cells (PC) would facilitate models for mechanistic studies and drug discovery, and also provide new cellular materials for regenerative therapies. A high-throughput chemical screen using CCS:lacZ and Contactin2:eGFP (Cntn2:eGFP) reporter embryonic stem cell (ESC) lines was used to discover a small molecule, sodium nitroprusside (SN), that efficiently promotes the generation of cardiac cells that express gene profiles and generate action potentials of PC-like cells. Imaging and mechanistic studies suggest that SN promotes the generation of PC from cardiac progenitors initially expressing cardiac myosin heavy chain, and that it does so by activating cAMP signaling. These findings provide a novel strategy to derive scalable PC, along with insight into the ontogeny of CCS development.

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Applications of miRNAs in cardiac development, disease progression and regeneration

Stem Cell Research & Therapy ◽

10.1186/s13287-019-1451-2 ◽

2019 ◽

Vol 10 (1) ◽

Cited By ~ 4

Author(s):

Jeremy Kah Sheng Pang ◽

Qian Hua Phua ◽

Boon-Seng Soh

Keyword(s):

Cardiac Development ◽

Control Cell ◽

Proliferative Potential ◽

Heart Cells ◽

Cardiovascular Development ◽

Developmental Defects ◽

Protein Coding ◽

Cardiac Progenitors ◽

Multiple Levels ◽

Cell Niche

AbstractDevelopment of the complex human heart is tightly regulated at multiple levels, maintaining multipotency and proliferative state in the embryonic cardiovascular progenitors and thereafter suppressing progenitor characteristics to allow for terminal differentiation and maturation. Small regulatory microRNAs (miRNAs) are at the level of post-transcriptional gene suppressors, which enhance the degradation or decay of their target protein-coding mRNAs. These miRNAs are known to play roles in a large number of biological events, cardiovascular development being no exception. A number of critical cardiac-specific miRNAs have been identified, of which structural developmental defects have been linked to dysregulation of miRNAs in the proliferating cardiac stem cells. These miRNAs present in the stem cell niche are lost when the cardiac progenitors terminally differentiate, resulting in the postnatal mitotic arrest of the heart. Therapeutic applications of these miRNAs extend to the realm of heart failure, whereby the death of heart cells in the ageing heart cannot be replaced due to the arrest of cell division. By utilizing miRNA therapy to control cell cycling, the regenerative potential of matured myocardium can be restored. This review will address the various cardiac progenitor-related miRNAs that control the development and proliferative potential of the heart.

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miR-300 mediates Bmi1 function and regulates differentiation in primitive cardiac progenitors

Cell Death and Disease ◽

10.1038/cddis.2015.255 ◽

2015 ◽

Vol 6 (10) ◽

pp. e1953-e1953 ◽

Cited By ~ 12

Author(s):

F M Cruz ◽

M Tomé ◽

J A Bernal ◽

A Bernad

Keyword(s):

Cardiac Progenitors

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Generation of Cardiomyocytes From Vascular Adventitia-Resident Stem Cells

Circulation Research ◽

10.1161/circresaha.117.312526 ◽

2018 ◽

Vol 123 (6) ◽

pp. 686-699 ◽

Cited By ~ 9

Author(s):

Subba Rao Mekala ◽

Philipp Wörsdörfer ◽

Jochen Bauer ◽

Olga Stoll ◽

Nicole Wagner ◽

...

Keyword(s):

Myocardial Infarction ◽

Heart Failure ◽

Stem Cells ◽

Endothelial Cells ◽

Aortic Wall ◽

Fluorescent Protein ◽

Therapeutic Option ◽

Cardiac Regeneration ◽

Cardiac Progenitors ◽

Resident Stem Cells

Rationale: Regeneration of lost cardiomyocytes is a fundamental unresolved problem leading to heart failure. Despite several strategies developed from intensive studies performed in the past decades, endogenous regeneration of heart tissue is still limited and presents a big challenge that needs to be overcome to serve as a successful therapeutic option for myocardial infarction. Objective: One of the essential prerequisites for cardiac regeneration is the identification of endogenous cardiomyocyte progenitors and their niche that can be targeted by new therapeutic approaches. In this context, we hypothesized that the vascular wall, which was shown to harbor different types of stem and progenitor cells, might serve as a source for cardiac progenitors. Methods and Results: We describe generation of spontaneously beating mouse aortic wall-derived cardiomyocytes without any genetic manipulation. Using aortic wall-derived cells (AoCs) of WT (wild type), αMHC (α-myosin heavy chain), and Flk1 (fetal liver kinase 1)-reporter mice and magnetic bead-associated cell sorting sorting of Flk1 + AoCs from GFP (green fluorescent protein) mice, we identified Flk1 + CD (cluster of differentiation) 34 + Sca-1 (stem cell antigen-1)-CD44 − AoCs as the population that gives rise to aortic wall-derived cardiomyocytes. This AoC subpopulation delivered also endothelial cells and macrophages with a particular accumulation within the aortic wall-derived cardiomyocyte containing colonies. In vivo, cardiomyocyte differentiation capacity was studied by implantation of fluorescently labeled AoCs into chick embryonic heart. These cells acquired cardiomyocyte-like phenotype as shown by αSRA (α-sarcomeric actinin) expression. Furthermore, coronary adventitial Flk1 + and CD34 + cells proliferated, migrated into the myocardium after mouse myocardial infarction, and expressed Isl-1 + (insulin gene enhancer protein-1) indicative of cardiovascular progenitor potential. Conclusions: Our data suggest Flk1 + CD34 + vascular adventitia-resident stem cells, including those of coronary adventitia, as a novel endogenous source for generating cardiomyocytes. This process is essentially supported by endothelial cells and macrophages. In summary, the therapeutic manipulation of coronary adventitia-resident cardiac stem and their supportive cells may open new avenues for promoting cardiac regeneration and repair after myocardial infarction and for preventing heart failure.

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Abstract 173: Clonal Analysis Reveals Limited Proliferative Capacity of Mature Cardiomyocytes during Embryonic Development.

Circulation Research ◽

10.1161/res.113.suppl_1.a173 ◽

2013 ◽

Vol 113 (suppl_1) ◽

Author(s):

Konstantina-Ioanna Sereti ◽

Paniz Kamran ◽

Shah R Ali ◽

Josh Z Lee ◽

Ali Subat ◽

...

Keyword(s):

Clonal Expansion ◽

Clonal Analysis ◽

Cardiac Cells ◽

Cell Labeling ◽

Postnatal Life ◽

Cardiovascular Development ◽

Reporter System ◽

Cellular Mechanisms ◽

Cardiac Progenitors ◽

Time Points

There is growing evidence that supports the ability of the heart to generate new cardiomyocytes during development and in response to injury albeit at a very low rate. It still remains unclear whether the newly generated cells originate from cardiac progenitor/stem cells or from pre-existing cardiomyocytes that re-enter the cell cycle. Understanding the cellular mechanisms regulating new cardiomyocyte formation is imperative towards the development of novel clinical strategies. We assessed the hypothesis that the cardiomyocytes generated during cardiac development are derived from cardiac progenitor cells and to a lesser extent form proliferating cardiomyocytes. We performed clonal analysis of cardiomyocyte formation by utilizing a stochastic multicolor reporter system (Rainbow) that allows random labeling of cells with three fluorescent proteins upon Cre-mediated recombination. Rainbow mice were crossed to αMHC-CreER mice for selective marking of cardiomyocytes or to Actin-CreER mice where all types of cells can be labeled. We induced rare recombination events at embryonic day E12.5 and clonal expansion of labeled cells was retrospectively analyzed at different time-points from E15.5 to postnatal day P30. To determine at which developmental stage cardiac cells lose their ability to proliferate, recombination was induced at different time-points from early embryonic to postnatal life and analysis was performed at P30. For each time-point, data was collected from ten mice. In αMHC-CreER;Rainbow mice, we observed mainly single cell labeling, a few doublets and some rare four-cell clones. At the same time-points, Actin-CreER;Rainbow hearts exhibited a large number of clones ranging from doublets to clones of more than twenty cells. Interestingly, clone formation and expansion was reduced with age. In conclusion, our results indicate that αMHC-positive cardiomyocyte proliferation is limited during development. On the other hand, non-αMHC expressing cells exhibited clonal expansion suggesting the possible contribution of cardiac progenitors even during late cardiovascular development.

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