Abstract 15756: Vascular Guidance of Cardiomyocyte Proliferation During Growth and Regeneration

Circulation ◽  
2020 ◽  
Vol 142 (Suppl_3) ◽  
Author(s):  
Paige DeBeneditts ◽  
Anish Karpurapu ◽  
Kyla Brezitski ◽  
Michael C Thomas ◽  
Ravi - Karra
Keyword(s):  
Large Scale ◽  
Nearest Neighbor ◽  
Critical Role ◽  
Cell Types ◽  
Neonatal Mouse ◽  
Angiogenic Factor ◽  
Heart Regeneration ◽  
Cardiomyocyte Proliferation ◽  
Cardiac Growth ◽  
Multiple Cell

Introduction: Stimulating cardiomyocyte (CM) proliferation is a major strategy for achieving therapeutic heart regeneration. However, heart regeneration requires coordinated interactions of multiple cell types. Because a hallmark of advanced heart failure is vascular rarefaction, the requirement of cardiac endothelial cells (CECs) for cardiac growth and regeneration is of particular importance. Hypothesis: We hypothesized that CECs are required for CM proliferation during growth and regeneration. Methods and Results: We performed a large-scale histologic assessment of neonatal mouse hearts and found the rate of CEC proliferation to shadow CM proliferation over the first 10 days of life. Using a nearest neighbor analysis, we found the fraction of proliferating CECs to be significantly enriched around cycling CMs compared to non-cycling CMs, suggesting that CEC and CM expansion are coupled within a myovascular niche. Single cell sequencing of neonatal mouse hearts after cryoinjury revealed that a majority of these proliferating CECs also express Vegfr2 . To functionally link CEC and CM proliferation, we generated Cdh5-CreER T2 ; Vefgr2 flox/flox mice to genetically delete Vegfr2 from CECs. Compared to mice with intact Vegfr2 , loss of Vegfr2 from CECs in neonatal mice leads to loss of CECs and severely dampens CM proliferation by 4 days (7.01±0.88% vs 0.39±0.35%, p = 7.4x10-4, n = 9),. Interestingly, CM proliferation is attenuated when Vegfr2 is deleted from CECs despite an increase of hypoxia indicators in CMs, signifying that hypoxia-induced CM proliferation is dependent on CECs. In contrast to CEC depletion, treatment of cryoinjured neonatal hearts with AAV encoding the master angiogenic factor, Vegfa can enhance heart regeneration with increased CM cycling in the borderzone (12.6±2.2% vs 5.4±0.4%, p = 0.02, n = 8), reduced scarring of the left ventricle (3.4±1.4% vs 7.6±1.2%, p = 03, n = 16), and improved fractional shortening (51.7±2.5% vs 36.7±4.3%, p = 0.007, n = 14). Conclusions: CEC and CM expansion are spatiotemporally coupled in a myovascular niche during cardiac growth. CECs play a critical role to support CM proliferation and are likely to provide instructive cues that may be leveraged for therapeutic heart regeneration.

scholarly journals Coupled myovascular expansion directs growth and regeneration of the neonatal mouse heart

2021 ◽  
Author(s):  
Paige DeBenedittis ◽  
Anish Karpurapu ◽  
Albert Henry ◽  
Michael C Thomas ◽  
Timothy J McCord ◽  
...  
Keyword(s):  
Cell Types ◽  
Neonatal Mouse ◽  
Border Zone ◽  
Mouse Heart ◽  
Heart Regeneration ◽  
Cardiac Growth ◽  
Spatial Coupling ◽  
Epicardial Cells ◽  
Multiple Cell ◽  
Vegfr2 Signaling

ABSTRACTInnate heart regeneration in zebrafish and neonatal mammals requires multiple cell types, such as epicardial cells, nerves, and macrophages, to enable proliferation of spared cardiomyocytes (CMs). How these cells interact to create growth niches is unclear. Here we profile proliferation kinetics of cardiac endothelial cells (CECs) and CMs in the neonatal mouse heart and find that CM and CEC expansion is spatiotemporally coupled. We show that coupled myovascular expansion during cardiac growth or regeneration is dependent upon VEGF-VEGFR2 signaling, as genetic deletion of Vegfr2 from CECs or inhibition of VEGFA abrogates both CEC and CM proliferation. Repair of cryoinjury, a model of incomplete regeneration, displays poor spatial coupling of CEC and CM proliferation. Boosting CEC density in the border zone by injection of virus encoding Vegfa enhances CM proliferation and the efficacy of heart regeneration, suggesting that revascularization strategies to increase CEC numbers may be an important adjunct for approaches designed to promote CM proliferation after injury. Finally, we use a human Mendelian randomization study to demonstrate that circulating VEGFA levels are positively associated with higher myocardial mass among healthy individuals, suggesting similar effects on human cardiac growth. Our work demonstrates the importance of coupled CEC and CM expansion for cardiomyogenesis and reveals the presence of a myovascular niche that underlies cardiac growth and regeneration.

scholarly journals Mechanisms linking adipose tissue inflammation to cardiac hypertrophy and fibrosis

2019 ◽  
Vol 133 (22) ◽  
pp. 2329-2344 ◽  
Author(s):  
Sarah R. Anthony ◽  
Adrienne R. Guarnieri ◽  
Anamarie Gozdiff ◽  
Robert N. Helsley ◽  
Albert Phillip Owens ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Cardiac Hypertrophy ◽  
Critical Role ◽  
Cell Types ◽  
Anatomical Location ◽  
Brown Adipocytes ◽  
Endocrine Organ ◽  
Inflammatory State ◽  
Multiple Cell ◽  
The Impact

Abstract Adipose tissue is classically recognized as the primary site of lipid storage, but in recent years has garnered appreciation for its broad role as an endocrine organ comprising multiple cell types whose collective secretome, termed as adipokines, is highly interdependent on metabolic homeostasis and inflammatory state. Anatomical location (e.g. visceral, subcutaneous, epicardial etc) and cellular composition of adipose tissue (e.g. white, beige, and brown adipocytes, macrophages etc.) also plays a critical role in determining its response to metabolic state, the resulting secretome, and its potential impact on remote tissues. Compared with other tissues, the heart has an extremely high and constant demand for energy generation, of which most is derived from oxidation of fatty acids. Availability of this fatty acid fuel source is dependent on adipose tissue, but evidence is mounting that adipose tissue plays a much broader role in cardiovascular physiology. In this review, we discuss the impact of the brown, subcutaneous, and visceral white, perivascular (PVAT), and epicardial adipose tissue (EAT) secretome on the development and progression of cardiovascular disease (CVD), with a particular focus on cardiac hypertrophy and fibrosis.

scholarly journals MUNIn (Multiple cell-type UNifying long-range chromatin Interaction detector): a statistical framework for identifying long-range chromatin interactions from multiple cell types

2020 ◽  
Author(s):  
Weifang Liu ◽  
Armen Abnousi ◽  
Qian Zhang ◽  
Yun Li ◽  
Ming Hu ◽  
...  
Keyword(s):  
Long Range ◽  
Spatial Organization ◽  
Critical Role ◽  
Cell Types ◽  
Chromatin Interaction ◽  
Cell Type ◽  
Statistical Framework ◽  
Chromatin Interactions ◽  
The Status ◽  
Multiple Cell

AbstractChromatin spatial organization (interactome) plays a critical role in genome function. Deep understanding of chromatin interactome can shed insights into transcriptional regulation mechanisms and human disease pathology. One essential task in the analysis of chromatin interactomic data is to identify long-range chromatin interactions. Existing approaches, such as HiCCUPS, FitHiC/FitHiC2 and FastHiC, are all designed for analyzing individual cell types. None of them accounts for unbalanced sequencing depths and heterogeneity among multiple cell types in a unified statistical framework. To fill in the gap, we have developed a novel statistical framework MUNIn (Multiple cell-type UNifying long-range chromatin Interaction detector) for identifying long-range chromatin interactions from multiple cell types. MUNIn adopts a hierarchical hidden Markov random field (H-HMRF) model, in which the status (peak or background) of each interacting chromatin loci pair depends not only on the status of loci pairs in its neighborhood region, but also on the status of the same loci pair in other cell types. To benchmark the performance of MUNIn, we performed comprehensive simulation studies and real data analysis, and showed that MUNIn can achieve much lower false positive rates for detecting cell-type-specific interactions (33.1 - 36.2%), and much enhanced statistical power for detecting shared peaks (up to 74.3%), compared to uni-cell-type analysis. Our data demonstrated that MUNIn is a useful tool for the integrative analysis of interactomic data from multiple cell types.

scholarly journals Runx1 promotes scar deposition and inhibits myocardial proliferation and survival during zebrafish heart regeneration

2019 ◽  
Author(s):  
Jana Koth ◽  
Xiaonan Wang ◽  
Abigail C. Killen ◽  
William T. Stockdale ◽  
Helen G. Potts ◽  
...  
Keyword(s):  
Plasminogen Activator Inhibitor ◽  
Cell Types ◽  
Degradation Pathway ◽  
Heart Regeneration ◽  
Injury Site ◽  
Heart Repair ◽  
Multiple Cell ◽  
Collagen Genes ◽  
Zebrafish Heart ◽  
Activator Inhibitor

Runx1 is a transcription factor that plays a key role in determining the proliferative and differential state of multiple cell-types, during both development and adulthood. Here, we report how runx1 is specifically upregulated at the injury site during zebrafish heart regeneration, but unexpectedly, absence of runx1 results in enhanced regeneration. Using single cell sequencing, we found that the wild-type injury site consists of Runx1-positive endocardial cells and thrombocytes that induce expression of smooth muscle and collagen genes without differentiating into myofibroblasts. Both these populations are absent in runx1 mutants, resulting in a less collagenous and fibrinous scar. The reduction in fibrin in the mutant is further explained by reduced myofibroblast formation and by upregulation of components of the fibrin degradation pathway, including plasminogen receptor Annexin 2A as well as downregulation of plasminogen activator inhibitor serpine1 in myocardium and endocardium, resulting in increased levels of Plasminogen. In addition, we find enhanced myocardial proliferation as well as increased myocardial survival in the mutant. Our findings suggest that Runx1 controls the regenerative response of multiple cardiac cell-types and that targeting Runx1 is a novel therapeutic strategy to induce endogenous heart repair.

scholarly journals Hydrogen Sulfide Promotes Cardiomyocyte Proliferation and Heart Regeneration via ROS Scavenging

2020 ◽  
Vol 2020 ◽  
pp. 1-11
Author(s):  
Jianqiu Pei ◽  
Fang Wang ◽  
Shengqiang Pei ◽  
Ruifeng Bai ◽  
Xiangfeng Cong ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Hydrogen Sulfide ◽  
Oxidative Dna Damage ◽  
Cardiac Injury ◽  
Protective Role ◽  
Neonatal Mouse ◽  
Heart Regeneration ◽  
Cardiomyocyte Proliferation ◽  
Ros Scavenging ◽  
Old Mice

Neonatal mouse hearts can regenerate completely in 21 days after cardiac injury, providing an ideal model to exploring heart regenerative therapeutic targets. The oxidative damage by Reactive Oxygen Species (ROS) is one of the critical reasons for the cell cycle arrest of cardiomyocytes (CMs), which cause mouse hearts losing the capacity to regenerate in 7 days or shorter after birth. As an antioxidant, hydrogen sulfide (H2S) plays a protective role in a variety of diseases by scavenging ROS produced during the pathological processes. In this study, we found that blocking H2S synthesis by PAG (H2S synthase inhibitor) suspended heart regeneration and CM proliferation with ROS deposition increase after cardiac injury (myocardial infarction or apex resection) in 2-day-old mice. NaHS (a H2S donor) administration improved heart regeneration with CM proliferation and ROS elimination after myocardial infarction in 7-day-old mice. NaHS protected primary neonatal mouse CMs from H2O2-induced apoptosis and promoted CM proliferation via SOD2-dependent ROS scavenging. The oxidative DNA damage in CMs was reduced with the elimination of ROS by H2S. Our results demonstrated for the first time that H2S promotes heart regeneration and identified NaHS as a potent modulator for cardiac repair.

scholarly journals Polyploidy in Cardiomyocytes

2020 ◽  
Vol 126 (4) ◽  
pp. 552-565 ◽  
Author(s):  
Wouter Derks ◽  
Olaf Bergmann
Keyword(s):  
Cardiac Injury ◽  
Physiological Role ◽  
Cell Types ◽  
Cell Number ◽  
The Body ◽  
Cardiomyocyte Hypertrophy ◽  
Heart Regeneration ◽  
Cardiomyocyte Proliferation ◽  
Hypertrophic Growth

The hallmark of most cardiac diseases is the progressive loss of cardiomyocytes. In the perinatal period, cardiomyocytes still proliferate, and the heart shows the capacity to regenerate upon injury. In the adult heart, however, the actual rate of cardiomyocyte renewal is too low to efficiently counteract substantial cell loss caused by cardiac injury. In mammals, cardiac growth by cell number expansion changes to growth by cardiomyocyte enlargement soon after birth, coinciding with a period in which most cardiomyocytes increase their DNA content by multinucleation and nuclear polyploidization. Although cardiomyocyte hypertrophy is often associated with these processes, whether polyploidy is a prerequisite or a consequence of hypertrophic growth is unclear. Both the benefits of cardiomyocyte enlargement over proliferative growth of the heart and the physiological role of polyploidy in cardiomyocytes are enigmatic. Interestingly, hearts in animal species with substantial cardiac regenerative capacity dominantly comprise diploid cardiomyocytes, raising the hypothesis that cardiomyocyte polyploidy poses a barrier for cardiomyocyte proliferation and subsequent heart regeneration. On the contrary, there is also evidence for self-duplication of multinucleated myocytes, suggesting a more complex picture of polyploidy in heart regeneration. Polyploidy is not restricted to the heart but also occurs in other cell types in the body. In this review, we explore the biological relevance of polyploidy in different species and tissues to acquire insight into its specific role in cardiomyocytes. Furthermore, we speculate about the physiological role of polyploidy in cardiomyocytes and how this might relate to renewal and regeneration.

scholarly journals Genome-wide CRISPR screen for PARKIN regulators reveals transcriptional repression as a determinant of mitophagy

2017 ◽  
Vol 115 (2) ◽  
pp. E180-E189 ◽  
Author(s):  
Christoph Potting ◽  
Christophe Crochemore ◽  
Francesca Moretti ◽  
Florian Nigsch ◽  
Isabel Schmidt ◽  
...  
Keyword(s):  
Transcriptional Repression ◽  
Critical Role ◽  
Induced Pluripotent Stem Cell ◽  
Cell Types ◽  
Physiological Regulation ◽  
Genome Wide ◽  
Crispr Screen ◽  
Multiple Cell ◽  
Induced Pluripotent ◽  
The Impact

PARKIN, an E3 ligase mutated in familial Parkinson’s disease, promotes mitophagy by ubiquitinating mitochondrial proteins for efficient engagement of the autophagy machinery. Specifically, PARKIN-synthesized ubiquitin chains represent targets for the PINK1 kinase generating phosphoS65-ubiquitin (pUb), which constitutes the mitophagy signal. Physiological regulation of PARKIN abundance, however, and the impact on pUb accumulation are poorly understood. Using cells designed to discover physiological regulators of PARKIN abundance, we performed a pooled genome-wide CRISPR/Cas9 knockout screen. Testing identified genes individually resulted in a list of 53 positive and negative regulators. A transcriptional repressor network including THAP11 was identified and negatively regulates endogenous PARKIN abundance. RNAseq analysis revealed the PARKIN-encoding locus as a prime THAP11 target, and THAP11 CRISPR knockout in multiple cell types enhanced pUb accumulation. Thus, our work demonstrates the critical role of PARKIN abundance, identifies regulating genes, and reveals a link between transcriptional repression and mitophagy, which is also apparent in human induced pluripotent stem cell-derived neurons, a disease-relevant cell type.

scholarly journals Heart regeneration: beyond new muscle and vessels

2021 ◽  
Author(s):  
Judy R Sayers ◽  
Paul R Riley
Keyword(s):  
Heart Development ◽  
Cell Function ◽  
Cell Types ◽  
Heart Regeneration ◽  
Cardiomyocyte Proliferation ◽  
Cardiovascular Tissue ◽  
Function Calls ◽  
Heart Muscle Cells ◽  
System Cell ◽  
Heart Injury

Abstract The most striking consequence of a heart attack is the loss of billions of heart muscle cells, alongside damage to the associated vasculature. The lost cardiovascular tissue is replaced by scar formation, which is non-functional and results in pathological remodelling of the heart and ultimately heart failure. It is, therefore, unsurprising that the heart regeneration field has centred efforts to generate new muscle and blood vessels through targeting cardiomyocyte proliferation and angiogenesis following injury. However, combined insights from embryological studies and regenerative models, alongside the adoption of -omics technology, highlight the extensive heterogeneity of cell types within the forming or re-forming heart and the significant crosstalk arising from non-muscle and non-vessel cells. In this review, we focus on the roles of fibroblasts, immune, conduction system, and nervous system cell populations during heart development and we consider the latest evidence supporting a function for these diverse lineages in contributing to regeneration following heart injury. We suggest that the emerging picture of neurologically, immunologically, and electrically coupled cell function calls for a wider-ranging combinatorial approach to heart regeneration.

Abstract 472: Cardiac Fibroblasts Regulate Myocardial Proliferation and Ventricular Formation through β1 Integrin Signaling

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Masaki Ieda ◽  
Takatoshi Tsuchihashi ◽  
Kathryn Ivey ◽  
Deepak Srivastava
Keyword(s):  
Extracellular Matrix ◽  
Cardiac Fibroblasts ◽  
Critical Role ◽  
Cell Types ◽  
Integrin Signaling ◽  
Β1 Integrin ◽  
Cardiomyocyte Proliferation ◽  
A Genome ◽  
Sirna Knockdown

Ventricular cardiomyocyte proliferation is essential to support increasing hemodynamic demands during late cardiogenesis. The increase in thickness of the ventricle is achieved by greater proliferation in the compact myocardium than in the trabecular myocardium. Disruption of this process leads to a disease known as ventricular non-compaction. Although epicardial-derived signals may contribute to the proliferative process, the factors and cell types responsible for development of the compact layer are unclear. In particular, the function of embryonic cardiac fibroblasts, derivatives from epicardium, and their secreted factors are largely unknown. By analyzing cell markers, we found that cardiac fibroblasts appeared in the compact myocardium at embryonic day 12.5 and increased in number coincident with growth of the compact myocardium. Using a novel co-culture system, we found that embryonic cardiac fibroblasts were much more effective in inducing cardiomyocyte proliferation than adult cardiac fibroblasts. A genome-wide screen and siRNA knockdown experiments revealed that fibronectin and collagen were embryonic cardiac fibroblast-specific signals that promoted cardiomyocyte proliferation through β1 integrin signaling in primary culture. In vivo, cardiomyocyte expression of β1 integrin was upregulated as cardiac fibroblasts appeared in the heart and secreted extracellular matrix proteins. In agreement with a critical role for β1 integrin, cardiomyocyte-specific deletion of β1 integrin in mice resulted in reduced myocardial proliferation and ventricular compaction, with downregulation of cyclinD1 and cyclinE1 and upregulation of cyclinG2 and p21, leading to prenatal death. These findings demonstrate that cardiac fibroblasts regulate cardiomyocyte proliferation and ventricular compaction through extracellular matrix/β1 integrin signaling.

scholarly journals DIANA-miRGen v4: indexing promoters and regulators for more than 1500 microRNAs

2020 ◽  
Vol 49 (D1) ◽  
pp. D151-D159
Author(s):  
Nikos Perdikopanis ◽  
Georgios K Georgakilas ◽  
Dimitris Grigoriadis ◽  
Vasilis Pierros ◽  
Ioannis Kavakiotis ◽  
...  
Keyword(s):  
Critical Role ◽  
Cell Types ◽  
Mirna Biogenesis ◽  
Transcriptional Level ◽  
Pathological State ◽  
Transcription Start Sites ◽  
Multiple Cell ◽  
Underlying Mechanisms ◽  
Cap Analysis ◽  
User Friendly

Abstract Deregulation of microRNA (miRNA) expression plays a critical role in the transition from a physiological to a pathological state. The accurate miRNA promoter identification in multiple cell types is a fundamental endeavor towards understanding and characterizing the underlying mechanisms of both physiological as well as pathological conditions. DIANA-miRGen v4 (www.microrna.gr/mirgenv4) provides cell type specific miRNA transcription start sites (TSSs) for over 1500 miRNAs retrieved from the analysis of >1000 cap analysis of gene expression (CAGE) samples corresponding to 133 tissues, cell lines and primary cells available in FANTOM repository. MiRNA TSS locations were associated with transcription factor binding site (TFBSs) annotation, for >280 TFs, derived from analyzing the majority of ENCODE ChIP-Seq datasets. For the first time, clusters of cell types having common miRNA TSSs are characterized and provided through a user friendly interface with multiple layers of customization. DIANA-miRGen v4 significantly improves our understanding of miRNA biogenesis regulation at the transcriptional level by providing a unique integration of high-quality annotations for hundreds of cell specific miRNA promoters with experimentally derived TFBSs.

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