Abstract 19324: Effects of Simulated Microgravity on Ckit+ Cardiac Progenitor Cells

Introduction: Space flight has profound negative impacts on cardiac health. Whereas microgravity appears to benefit cardiomyogenesis, long-duration space flight results in increased risk for cardiomyopathy. Here, we focused on cKit+ cardiac progenitor cells (CPCs) to elucidate the effects of microgravity in the heart. Hypothesis: Microgravity inhibits migration, proliferation and differentiation of CPCs. Methods: Adult heart tissue or induced pluripotent stem cells (iPSCs) from cKitCreErt2;IRG mice were grown for up to 24- (n=5) or 21-days (n=6), respectively, in static (SC) or a rotary cell-culture system (RCCS, simulated microgravity) in the presence of 4-OH tamoxifen to irreversibly label CPCs with EGFP. Expression of EGFP was quantified at selected time points in heart explants and iPSC-derived beating embryoid bodies (EBs). In addition, microarray analysis was performed on EBs at selected time points (n=11). Results: We found that, although explants in SC consistently produced EGFP+ CPCs with full capacity to proliferate and migrate, expression of EGFP was abolished in RCCS (p<0.05). Similarly, when day-4 EBs (formed via the hanging-drop method) were transferred to RCCS, they generated significantly fewer spontaneously beating EBs compared to EBs grown in SC (p=0.0005), whereas expression of EGFP in beating EBs was downregulated ~10-fold (p=0.01). Microarray analysis of EBs illustrated that the effect of CPs was accompanied by downregulation of genes related to migration, differentiation and development of the cardiac neural crest cell (CNC) lineage (i.e. Pax3, semaphorins, endothelin) without affecting the expression of cardiac mesoderm-related genes (i.e. GATA4, NKX2-5, MEF2C). Intriguingly, the effect of RCCS in CNC-related genes could be partly rescued upon transfer of EBs from RCCS to SC. Conclusions: cKit expression and CNC pathways are inhibited under simulated microgravity but can be reversed by returning to normal gravity. Our findings provide novel insights into the role of gravity in cardiomyogenesis and suggest that CPCs should be targeted therapeutically for the prevention and treatment of microgravity-induced cardiomyopathy.

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

Circulation Research ◽

10.1161/res.119.suppl_1.323 ◽

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.

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Effects of Spaceflight and Simulated Microgravity on YAP1 Expression in Cardiovascular Progenitors: Implications for Cell-Based Repair

International Journal of Molecular Sciences ◽

10.3390/ijms20112742 ◽

2019 ◽

Vol 20 (11) ◽

pp. 2742 ◽

Cited By ~ 8

Author(s):

Victor Camberos ◽

Jonathan Baio ◽

Leonard Bailey ◽

Nahidh Hasaniya ◽

Larry V. Lopez ◽

...

Keyword(s):

Progenitor Cells ◽

Simulated Microgravity ◽

Cardiac Development ◽

Hippo Pathway ◽

Space Station ◽

Cardiac Regeneration ◽

Cardiac Progenitor Cells ◽

Hippo Signaling ◽

Sheep Model ◽

The Hippo Pathway

Spaceflight alters many processes of the human body including cardiac function and cardiac progenitor cell behavior. The mechanism behind these changes remains largely unknown; however, simulated microgravity devices are making it easier for researchers to study the effects of microgravity. To study the changes that take place in cardiac progenitor cells in microgravity environments, adult cardiac progenitor cells were cultured aboard the International Space Station (ISS) as well as on a clinostat and examined for changes in Hippo signaling, a pathway known to regulate cardiac development. Cells cultured under microgravity conditions, spaceflight-induced or simulated, displayed upregulation of downstream genes involved in the Hippo pathway such as YAP1 and SOD2. YAP1 is known to play a role in cardiac regeneration which led us to investigate YAP1 expression in a sheep model of cardiovascular repair. Additionally, to mimic the effects of microgravity, drug treatment was used to induce Hippo related genes as well as a regulator of the Hippo pathway, miRNA-302a. These studies provide insight into the changes that occur in space and how the effects of these changes relate to cardiac regeneration studies.

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Microarray analysis of genes differentially expressed in hepatoblastoma cell line cultured in simulated microgravity

Journal of Hepatology ◽

10.1016/s0168-8278(01)80279-8 ◽

2001 ◽

Vol 34 (0) ◽

pp. 80

Author(s):

V Khaoustov

Keyword(s):

Cell Line ◽

Microarray Analysis ◽

Simulated Microgravity ◽

Differentially Expressed ◽

Hepatoblastoma Cell Line

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Cardiac progenitor cells in terminally failing hearts: Immunohistological observations in human myocardium

The Thoracic and Cardiovascular Surgeon ◽

10.1055/s-2007-967414 ◽

2007 ◽

Vol 55 (S 1) ◽

Author(s):

M Arnold ◽

V Kufer ◽

A Schütz ◽

B Reiter ◽

M Fittkau ◽

...

Keyword(s):

Progenitor Cells ◽

Human Myocardium ◽

Cardiac Progenitor Cells

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Breakthrough Regenerative Cardiopoietic Stem Cells and Related Technologies in the Field of Cardiovascular Medicine – From Bench to Bedside

Interventional Cardiology Review ◽

10.15420/icr.2012.7.1.14 ◽

2012 ◽

Vol 7 (1) ◽

pp. 14

Author(s):

Christian Homsy ◽

Keyword(s):

Stem Cells ◽

Cell Therapy ◽

Progenitor Cells ◽

Cardiac Disease ◽

Cardiac Tissue ◽

Cardiac Repair ◽

Cardiac Diseases ◽

Cardiac Progenitor Cells ◽

Biotechnology Company ◽

Cardiovascular Medicine

The scale of cardiac diseases, and in particular heart failure and acute myocardial infarction, emphasises the need for radically new approaches, such as cell therapy, to address the underlying cause of the disease, the loss of functional myocardium. Stem cell-based therapies, whether through transplanted cells or directing innate repair, may provide regenerative approaches to cardiac diseases by halting, or even reversing, the events responsible for progression of organ failure. Cardio3 BioSciences, a leading Belgian biotechnology company focused on the discovery and development of regenerative and protective therapies for the treatment of cardiac disease, was founded in this context in 2004. The company is developing a highly innovative cell therapy approach based on a platform designed to reprogramme the patient’s own stem cells into cardiac progenitor cells. The underlying rationale behind this approach is that, in order to reconstruct cardiac tissue, stem cells need to be specific to cardiac tissue. The key is therefore to provide cardiac-specific progenitor cells to the failing heart to induce cardiac repair.

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Circulating Stem/Progenitor Cells as Prognostic Biomarkers in Macro- and Microvascular Disease: A Narrative Review of Prospective Observational Studies

Current Medicinal Chemistry ◽

10.2174/0929867324666170920154020 ◽

2018 ◽

Vol 25 (35) ◽

pp. 4507-4517 ◽

Cited By ~ 6

Author(s):

Mauro Rigato ◽

Gian Paolo Fadini

Keyword(s):

Cardiovascular Disease ◽

Progenitor Cells ◽

Cardiovascular Events ◽

Observational Studies ◽

English Language ◽

Microvascular Disease ◽

Patient Characteristics ◽

Diabetic Patients ◽

Increased Risk ◽

Cell Phenotypes

Background: Circulating progenitor cells (CPCs) and endothelial progenitor cells (EPCs) are immature cells involved in vascular repair and related to many aspects of macro and microvascular disease. <p> Objective: We aimed to review studies reporting the prognostic role of CPCs/EPCs measurement on development of cardiovascular disease and microangiopathy. <p> Methods and Results: We reviewed the English language literature for prospective observational studies reporting the future development of cardiovascular disease or microangiopathy in patients having a baseline determination of CPCs/EPCs. We retrieved 34 studied reporting on cardiovascular outcomes and 2 studies reporting on microvascular outcomes. Overall, a reduced baseline level of CPCs/EPCs was associated with a significant increased risk of cardiovascular events, all-cause death, and onset/progression of microangiopathy. The most predictive phenotypes were CD34+ and CD34+CD133+. The main limitation was related to the high heterogeneity among studies in terms of patient characteristics and cell phenotypes. <p> Conclusion: The present review shows that a reduced level of circulating progenitor cells is a risk factor for the development of future cardiovascular events and death. In addition, low CPCs/EPCs levels predict the onset or worsening of microalbuminuria and retinopathy in diabetic patients.

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Diabetes induces dysregulation of microRNAs associated with survival, proliferation and self-renewal in cardiac progenitor cells

Diabetologia ◽

10.1007/s00125-021-05405-7 ◽

2021 ◽

Author(s):

Nima Purvis ◽

Sweta Kumari ◽

Dhananjie Chandrasekera ◽

Jayanthi Bellae Papannarao ◽

Sophie Gandhi ◽

...

Keyword(s):

Progenitor Cells ◽

Cardiac Progenitor Cells ◽

Self Renewal

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Novel Identified Circular Transcript of RCAN2, circ-RCAN2, Shows Deviated Expression Pattern in Pig Reperfused Infarcted Myocardium and Hypoxic Porcine Cardiac Progenitor Cells In Vitro

International Journal of Molecular Sciences ◽

10.3390/ijms22031390 ◽

2021 ◽

Vol 22 (3) ◽

pp. 1390

Author(s):

Julia Mester-Tonczar ◽

Patrick Einzinger ◽

Johannes Winkler ◽

Nina Kastner ◽

Andreas Spannbauer ◽

...

Keyword(s):

Myocardial Infarction ◽

Progenitor Cells ◽

Regulatory Networks ◽

Circular Rnas ◽

Cardiac Progenitor Cells ◽

Tissue Samples ◽

Healthy Myocardium ◽

Acute Mi

Circular RNAs (circRNAs) are crucial in gene regulatory networks and disease development, yet circRNA expression in myocardial infarction (MI) is poorly understood. Here, we harvested myocardium samples from domestic pigs 3 days after closed-chest reperfused MI or sham surgery. Cardiac circRNAs were identified by RNA-sequencing of rRNA-depleted RNA from infarcted and healthy myocardium tissue samples. Bioinformatics analysis was performed using the CIRIfull and KNIFE algorithms, and circRNAs identified with both algorithms were subjected to differential expression (DE) analysis and validation by qPCR. Circ-RCAN2 and circ-C12orf29 expressions were significantly downregulated in infarcted tissue compared to healthy pig heart. Sanger sequencing was performed to identify the backsplice junctions of circular transcripts. Finally, we compared the expressions of circ-C12orf29 and circ-RCAN2 between porcine cardiac progenitor cells (pCPCs) that were incubated in a hypoxia chamber for different time periods versus normoxic pCPCs. Circ-C12orf29 did not show significant DE in vitro, whereas circ-RCAN2 exhibited significant ischemia-time-dependent upregulation in hypoxic pCPCs. Overall, our results revealed novel cardiac circRNAs with DE patterns in pCPCs, and in infarcted and healthy myocardium. Circ-RCAN2 exhibited differential regulation by myocardial infarction in vivo and by hypoxia in vitro. These results will improve our understanding of circRNA regulation during acute MI.

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