44 results found
McCracken IR, Saginc G, He L, et al., 2021, Lack of evidence of ACE2 expression and replicative infection by SARSCoV-2 in human endothelial cells, Circulation, Vol: 143, Pages: 865-868, ISSN: 0009-7322
Cardiovascular disease is the leading cause of death worldwide. Advanced insights into disease mechanisms and therapeutic strategies require deeper understanding of the healthy heart’s molecular processes. Knowledge of the full repertoire of cardiac cells and their gene expression profiles is a fundamental first step in this endeavor. Here, using state-of-the-art analyses of large-scale single-cell and nuclei transcriptomes, we characterise six anatomical adult heart regions. Our results highlight the cellular heterogeneity of cardiomyocytes, pericytes, and fibroblasts, revealing distinct atrial and ventricular subsets with diverse developmental origins and specialized properties. We define the complexity of the cardiac vasculature and its changes along the arterio-venous axis. In the immune compartment we identify cardiac resident macrophages with inflammatory and protective transcriptional signatures. Further, inference of cell-cell interactions highlight different macrophage-fibroblast-cardiomyocyte networks between atria and ventricles that are distinct from skeletal muscle. Our human cardiac cell atlas improves our understanding of the human heart and provides a healthy reference for future studies.
Constantinou C, Miranda Almeida A, Chaves Guerrero P, et al., 2020, Human pluripotent stem cell-derived cardiomyocytes as a targetplatform for paracrine protection by cardiac mesenchymal stromalcells, Scientific Reports, Vol: 10, ISSN: 2045-2322
Ischemic heart disease remains the foremost cause of death globally, with survivors at risk for subsequent heart failure. Paradoxically, cell therapies to offset cardiomyocyte loss after ischemic injury improve long-term cardiac function despite a lack of durable engraftment. An evolving consensus, inferred preponderantly from non-human models, is that transplanted cells benefit the heart via early paracrinesignals. Here, we tested the impact of paracrine signals on human cardiomyocytes, using human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) as the target of mouse and human cardiac mesenchymal stromal cells (cMSC) with progenitor-like features. In co-culture and conditioned medium studies, cMSCs markedly inhibited human cardiomyocyte death. Little or no protection was conferred by mouse tail tip or human skin fibroblasts. Consistent with the results of transcriptomic profiling, functional analyses showed that the cMSC secretome suppressed apoptosis and and preserved cardiac mitochondrial transmembrane potential. Protection was independent of exosomes under the conditions tested. In mice, injecting cMSC-conditioned media into the infarct border zone reduced apoptotic cardiomyocytes >70% locally. Thus, hPSC-CMs provide an auspicious, relevant human platform to investigate extracellular signals for cardiac muscle survival, substantiating human cardioprotection by cMSCs, and suggesting the cMSC secretome or its components as potential cell-free therapeutic products.
Sungnak W, Huang N, Bécavin C, et al., 2020, SARS-CoV-2 entry factors are highly expressed in nasal epithelial cells together with innate immune genes., Nat Med, Vol: 26, Pages: 681-687
We investigated SARS-CoV-2 potential tropism by surveying expression of viral entry-associated genes in single-cell RNA-sequencing data from multiple tissues from healthy human donors. We co-detected these transcripts in specific respiratory, corneal and intestinal epithelial cells, potentially explaining the high efficiency of SARS-CoV-2 transmission. These genes are co-expressed in nasal epithelial cells with genes involved in innate immunity, highlighting the cells' potential role in initial viral infection, spread and clearance. The study offers a useful resource for further lines of inquiry with valuable clinical samples from COVID-19 patients and we provide our data in a comprehensive, open and user-friendly fashion at www.covid19cellatlas.org.
Fiedler LR, Chapman K, Xie M, et al., 2020, MAP4K4 Inhibition Promotes Survival of Human Stem Cell-Derived Cardiomyocytes and Reduces Infarct Size In Vivo (vol 24, 579.e1,2019), CELL STEM CELL, Vol: 26, Pages: 458-458, ISSN: 1934-5909
Constantinou C, Noseda M, Chaves P, et al., 2019, Human Cardiomyocyte Protection From Apoptosis by Cardiac Progenitor Cell Secreted Factors, Scientific Sessions of the American-Heart-Association, Publisher: LIPPINCOTT WILLIAMS & WILKINS, ISSN: 0009-7322
Schneider M, Fiedler L, Chapman K, et al., 2019, MAP4K4 inhibition promotes survival of human stem cell derived cardiomyocyte and reduces infarct size in vivo, Cell Stem Cell, Vol: 24, Pages: 579-591.e12, ISSN: 1875-9777
Heart disease is a paramount cause of global death and disability. Although cardiomyocyte death plays a causal role and its suppression would be logical, no clinical counter-measures target the responsible intracellular pathways. Therapeutic progress has been hampered by lack of preclinical human validation. Mitogen-activated protein kinase kinase kinase kinase-4 (MAP4K4) is activated in failing human hearts and relevant rodent models. Using human induced-pluripotent-stem-cell-derived cardiomyocytes (hiPSC-CMs) and MAP4K4 gene silencing, we demonstrate that death induced by oxidative stress requires MAP4K4. Consequently, we devised a small-molecule inhibitor, DMX-5804, that rescues cell survival, mitochondrial function, and calcium cycling in hiPSC-CMs. As proof of principle that drug discovery in hiPSC-CMs may predict efficacy in vivo, DMX-5804 reduces ischemia-reperfusion injury in mice by more than 50%. We implicate MAP4K4 as a well-posed target toward suppressing human cardiac cell death and highlight the utility of hiPSC-CMs in drug discovery to enhance cardiomyocyte survival.
Noseda M, Samari S, Chaves P, et al., 2019, Modulation of Macrophage Differentiation and Activation: Paracrine Signals from Cardiac Progenitor Cells, Publisher: SPRINGER, Pages: 262-262, ISSN: 0920-3206
Massaia A, Chaves P, Samari S, et al., 2018, Single cell gene expression to understand the dynamic architecture of the heart, Frontiers in Cardiovascular Medicine, Vol: 5, ISSN: 2297-055X
The recent development of single cell gene expression technologies, and especially singlecell transcriptomics, have revolutionized the way biologists and clinicians investigateorgans and organisms, allowing an unprecedented level of resolution to the descriptionof cell demographics in both healthy and diseased states. Single cell transcriptomicsprovide information on prevalence, heterogeneity, and gene co-expression at theindividual cell level. This enables a cell-centric outlook to define intracellular generegulatory networks and to bridge toward the definition of intercellular pathwaysotherwise masked in bulk analysis. The technologies have developed at a fast paceproducing a multitude of different approaches, with several alternatives to choose fromat any step, including single cell isolation and capturing, lysis, RNA reverse transcriptionand cDNA amplification, library preparation, sequencing, and computational analyses.Here, we provide guidelines for the experimental design of single cell RNA sequencingexperiments, exploring the current options for the crucial steps. Furthermore, we providea complete overview of the typical data analysis workflow, from handling the rawsequencing data to making biological inferences. Significantly, advancements in singlecell transcriptomics have already contributed to outstanding exploratory and functionalstudies of cardiac development and disease models, as summarized in this review. Inconclusion, we discuss achievable outcomes of single cell transcriptomics’ applicationsin addressing unanswered questions and influencing future cardiac clinical applications.
Shanmuganathan M, Vughs J, Noseda M, et al., 2018, Exosomes: Basic biology and technological advancements suggesting their potential as ischemic heart disease therapeutics, Frontiers in Physiology, Vol: 9, ISSN: 1664-042X
Exosomes are small nano-sized vesicles that deliver biologically active RNA molecules and proteins to recipient cells through binding, fusion or endocytosis. There is emerging evidence that endogenous exosomes released by cardiovascular cells and progenitor cells impact cell survival and proliferation, thus regulating angiogenesis, cardiac protection and repair. These cardioprotective and regenerative traits have the potential to translate in to novel therapeutic options for post-ischaemic cardiac regeneration, thus potentially delaying the progression to ischaemic heart failure. Cellular stressors influence exosomes' secretion and the molecular composition of the exosome cargo, thus impacting on the above processes. Evidences are emerging that loading of proteins and RNAs in the exosomes cargos can be manipulated. Similarly, manipulation of exosomes surface proteins' expression to target exosomes to specific cells and tissues is doable. In addition, nature-inspired synthetic exosomes can be assembled to deliver specific clues to the recipient cells, including proliferative and differentiation stimuli, or shed paracrine signals enabling to reconstructing the heart homeostatic micro-environment. This review will describe exosome biogenesis and emerging evidence of exosome-mediated regenerative cell-to-cell communications and will conclude discussing possibilities of using exosomes to treat ischemic heart disease.
Noseda M, Harding SE, 2018, Understanding dynamic tissue organization by studying the human body one cell at a time: the human cell atlas (HCA) project, CARDIOVASCULAR RESEARCH, Vol: 114, Pages: E93-E95, ISSN: 0008-6363
Adamowicz M, Morgan CC, Haubner BJ, et al., 2018, Functionally conserved noncoding regulators of cardiomyocyte proliferation and regeneration in mouse and human, Circulation: Cardiovascular Genetics, Vol: 11, ISSN: 1942-325X
Background: The adult mammalian heart has little regenerative capacity after myocardial infarction (MI), whereas neonatal mouse heart regenerates without scarring or dysfunction. However, the underlying pathways are poorly defined. We sought to derive insights into the pathways regulating neonatal development of the mouse heart and cardiac regeneration post-MI.Methods and Results: Total RNA-seq of mouse heart through the first 10 days of postnatal life (referred to as P3, P5, P10) revealed a previously unobserved transition in microRNA (miRNA) expression between P3 and P5 associated specifically with altered expression of protein-coding genes on the focal adhesion pathway and cessation of cardiomyocyte cell division. We found profound changes in the coding and noncoding transcriptome after neonatal MI, with evidence of essentially complete healing by P10. Over two-thirds of each of the messenger RNAs, long noncoding RNAs, and miRNAs that were differentially expressed in the post-MI heart were differentially expressed during normal postnatal development, suggesting a common regulatory pathway for normal cardiac development and post-MI cardiac regeneration. We selected exemplars of miRNAs implicated in our data set as regulators of cardiomyocyte proliferation. Several of these showed evidence of a functional influence on mouse cardiomyocyte cell division. In addition, a subset of these miRNAs, miR-144-3p, miR-195a-5p, miR-451a, and miR-6240 showed evidence of functional conservation in human cardiomyocytes.Conclusions: The sets of messenger RNAs, miRNAs, and long noncoding RNAs that we report here merit further investigation as gatekeepers of cell division in the postnatal heart and as targets for extension of the period of cardiac regeneration beyond the neonatal period.
Noseda M, Constantinou C, Samari S, et al., 2017, Abstract 191: Paracrine Impact of Cardiac Progenitor Cells on Macrophage Phenotypes and Human Ipsc-derived Cardiomyocyte Survival, Basic Cardiovascular Sciences Scientific Sessions of the American-Heart-Association - Pathways to Cardiovascular Therapeutics, Publisher: LIPPINCOTT WILLIAMS & WILKINS, ISSN: 0009-7330
Speidel AT, Stuckey DJ, Chow LW, et al., 2017, Multi-modal hydrogel-based platform to deliver and monitor cardiac progenitor/stem cell engraftment, ACS Central Science, Vol: 3, Pages: 338-348, ISSN: 2374-7951
Retention and survival of transplanted cells are major limitations to the efficacy of regenerative medicine, with short-term paracrine signals being the principal mechanism underlying current cell therapies for heart repair. Consequently, even improvements in short-term durability may have a potential impact on cardiac cell grafting. We have developed a multimodal hydrogel-based platform comprised of a poly(ethylene glycol) network cross-linked with bioactive peptides functionalized with Gd(III) in order to monitor the localization and retention of the hydrogel in vivo by magnetic resonance imaging. In this study, we have tailored the material for cardiac applications through the inclusion of a heparin-binding peptide (HBP) sequence in the cross-linker design and formulated the gel to display mechanical properties resembling those of cardiac tissue. Luciferase-expressing cardiac stem cells (CSC-Luc2) encapsulated within these gels maintained their metabolic activity for up to 14 days in vitro. Encapsulation in the HBP hydrogels improved CSC-Luc2 retention in the mouse myocardium and hind limbs at 3 days by 6.5- and 12- fold, respectively. Thus, this novel heparin-binding based, Gd(III)-tagged hydrogel and CSC-Luc2 platform system demonstrates a tailored, in vivo detectable theranostic cell delivery system that can be implemented to monitor and assess the transplanted material and cell retention.
Wei K, Serpooshan V, Hurtado C, et al., 2015, Epicardial FSTL1 reconstitution regenerates the adult mammalian heart, Nature, Vol: 525, Pages: 479-485, ISSN: 0028-0836
The elucidation of factors that activate the regeneration of the adult mammalian heart is of major scientific and therapeutic importance. Here we found that epicardial cells contain a potent cardiogenic activity identified as follistatin-like 1 (Fstl1). Epicardial Fstl1 declines following myocardial infarction and is replaced by myocardial expression. Myocardial Fstl1 does not promote regeneration, either basally or upon transgenic overexpression. Application of the human Fstl1 protein (FSTL1) via an epicardial patch stimulates cell cycle entry and division of pre-existing cardiomyocytes, improving cardiac function and survival in mouse and swine models of myocardial infarction. The data suggest that the loss of epicardial FSTL1 is a maladaptive response to injury, and that its restoration would be an effective way to reverse myocardial death and remodelling following myocardial infarction in humans.
Morez CY, Noseda M, Abreu Paiva M, et al., 2015, Enhanced efficiency of genetic programming toward cardiomyocyte creation through topographical cues, Biomaterials, Vol: 70, Pages: 94-104, ISSN: 1878-5905
Generation of de novo cardiomyocytes through viral over-expression of key transcription factors represents a highly promising strategy for cardiac muscle tissue regeneration. Although the feasibility of cell reprogramming has proven possible both in vitro and in vivo, the efficiency of the process remains extremely low. Here, we report a chemical-free technique in which topographical cues, more specifically parallel microgrooves, enhance the trans-differentiation of cardiac progenitors into cardiomyocyte-like cells. Using a lentivirus-mediated direct reprogramming strategy for expression of Myocardin, Tbx5, and Mef2c, we showed that the microgrooved substrate provokes an increase in histone H3 acetylation (AcH3), known to be a permissive environment for reprogramming by “stemness” factors, as well as stimulation of myocardin sumoylation, a post-translational modification essential to the transcriptional function of this key co-activator. These biochemical effects mimicked those of a pharmacological histone deacetylase inhibitor, valproic acid (VPA), and like VPA markedly augmented the expression of cardiomyocyte-specific proteins by the genetically engineered cells. No instructive effect was seen in cells unresponsive to VPA. In addition, the anisotropy resulting from parallel microgrooves induced cellular alignment, mimicking the native ventricular myocardium and augmenting sarcomere organization.
Harada M, Noseda M, Schneider M, 2015, The Cre / Lox P fate mapping study on cardiac progenitor cells revealed its embryonic origin with its intrinsic methodological limitation validated by single cell qRT-PCR, Congress of the European-Society-of-Cardiology (ESC), Publisher: OXFORD UNIV PRESS, Pages: 950-950, ISSN: 0195-668X
Noseda M, Abreu-Paiva M, Schneider MD, 2015, The Quest for the Adult Cardiac Stem Cell, Circulation Journal, Vol: 79, Pages: 1422-1430, ISSN: 1347-4820
Over the past 2 decades, cardiac regeneration has evolved from an exotic fringe of cardiovascular biology to theforefront of molecular, genetic, epigenetic, translational, and clinical investigations. The unmet patient need is thepaucity of self-repair following infarction. Robust regeneration seen in models such as zebrafish and newborn micehas inspired the field, along with encouragement from modern methods that make even low levels of restorativegrowth discernible, changing the scientific and technical landscape for effective counter-measures. Approachesunder study to augment cardiac repair complement each other, and encompass grafting cells of diverse kinds,restarting the cell cycle in post-mitotic ventricular myocytes, reprogramming non-myocytes, and exploiting the dormantprogenitor/stem cells that lurk within the adult heart. The latter are the emphasis of the present review. Cardiacresidentstem cells (CSC) can be harvested from heart tissue, expanded, and delivered to the myocardium as atherapeutic product, whose benefits may be hoped to surpass those achieved in human trials of bone marrow.However, important questions are prompted by such cells’ discovery. How do they benefit recipient hearts? Do theycontribute, measurably, as an endogenous population, to self-repair? Even if “no,” might CSCs be targets for activationin situ by growth factors and other developmental catalysts? And, what combination of distinguishing markersbest demarcates the cells with robust clonal growth and cardiogenic potential?
Belian E, Noseda M, Abreu Paiva MS, et al., 2015, Forward Programming of Cardiac Stem Cells by Homogeneous Transduction with MYOCD plus TBX5., PLOS One, Vol: 10, ISSN: 1932-6203
UNLABELLED: Adult cardiac stem cells (CSCs) express many endogenous cardiogenic transcription factors including members of the Gata, Hand, Mef2, and T-box family. Unlike its DNA-binding targets, Myocardin (Myocd)-a co-activator not only for serum response factor, but also for Gata4 and Tbx5-is not expressed in CSCs. We hypothesised that its absence was a limiting factor for reprogramming. Here, we sought to investigate the susceptibility of adult mouse Sca1+ side population CSCs to reprogramming by supplementing the triad of GATA4, MEF2C, and TBX5 (GMT), and more specifically by testing the effect of the missing co-activator, Myocd. Exogenous factors were expressed via doxycycline-inducible lentiviral vectors in various combinations. High throughput quantitative RT-PCR was used to test expression of 29 cardiac lineage markers two weeks post-induction. GMT induced more than half the analysed cardiac transcripts. However, no protein was detected for the induced sarcomeric genes Actc1, Myh6, and Myl2. Adding MYOCD to GMT affected only slightly the breadth and level of gene induction, but, importantly, triggered expression of all three proteins examined (α-cardiac actin, atrial natriuretic peptide, sarcomeric myosin heavy chains). MYOCD + TBX was the most effective pairwise combination in this system. In clonal derivatives homogenously expressing MYOCD + TBX at high levels, 93% of cardiac transcripts were up-regulated and all five proteins tested were visualized. IN SUMMARY: (1) GMT induced cardiac genes in CSCs, but not cardiac proteins under the conditions used. (2) Complementing GMT with MYOCD induced cardiac protein expression, indicating a more complete cardiac differentiation program. (3) Homogeneous transduction with MYOCD + TBX5 facilitated the identification of differentiating cells and the validation of this combinatorial reprogramming strategy. Together, these results highlight the pivotal importance of MYOCD in driving CSCs toward a cardiac muscle fate
Noseda M, Harada M, McSweeney S, et al., 2015, PDGFRα demarcates the cardiogenic clonogenic Sca1(+) stem/progenitor cell in adult murine myocardium, Nature Communications, Vol: 6, ISSN: 2041-1723
Cardiac progenitor/stem cells in adult hearts represent an attractive therapeutic target for heart regeneration, though (inter)-relationships among reported cells remain obscure. Using single-cell qRT-PCR and clonal analyses, here we define four subpopulations of cardiac progenitor/stem cells in adult mouse myocardium all sharing stem cell antigen-1 (Sca1), based on side population (SP) phenotype, PECAM-1 (CD31) and platelet-derived growth factor receptor-α (PDGFRα) expression. SP status predicts clonogenicity and cardiogenic gene expression (Gata4/6, Hand2 and Tbx5/20), properties segregating more specifically to PDGFRα(+) cells. Clonal progeny of single Sca1(+) SP cells show cardiomyocyte, endothelial and smooth muscle lineage potential after cardiac grafting, augmenting cardiac function although durable engraftment is rare. PDGFRα(-) cells are characterized by Kdr/Flk1, Cdh5, CD31 and lack of clonogenicity. PDGFRα(+)/CD31(-) cells derive from cells formerly expressing Mesp1, Nkx2-5, Isl1, Gata5 and Wt1, distinct from PDGFRα(-)/CD31(+) cells (Gata5 low; Flk1 and Tie2 high). Thus, PDGFRα demarcates the clonogenic cardiogenic Sca1(+) stem/progenitor cell.
Noseda M, Harada M, Mcsweeney S, et al., 2014, PDGFRalpha demarcates the cardiogenic and clonogenic Sca-1+stem cell, 3rd Congress of the ESC-Council-on-Basic-Cardiovascular-Science on Frontiers in Cardio Vascular Biology, Publisher: OXFORD UNIV PRESS, ISSN: 0008-6363
Belian E, Noseda M, Leja T, et al., 2012, Is Myocardin a limiting factor for cardiac programming?, Scientific Sessions of the American-Heart-Association, Publisher: LIPPINCOTT WILLIAMS & WILKINS, Pages: E386-E386, ISSN: 0009-7330
Noseda M, De Smith AJ, Leia T, et al., 2012, The Cdkn2a/2b Tumour Suppressor Locus is a Hot Spot for Genome Instability in Mouse Cardiac Progenitor Cells, CIRCULATION, Vol: 126, ISSN: 0009-7322
Noseda M, Harada M, McSweeney SJ, et al., 2012, Unmasking Phenotypic Micro-heterogeneities in Adult Cardiac Progenitor Cells: Clonal Analysis, Single Cell QRT-PCR, and Fate Mapping, CIRCULATION, Vol: 126, ISSN: 0009-7322
Chang L, Noseda M, Higginson M, et al., 2012, Differentiation of vascular smooth muscle cells from local precursors during embryonic and adult arteriogenesis requires Notch signaling., Proc Natl Acad Sci U S A, Vol: 109, Pages: 6993-6998
Vascular smooth muscle cells (VSMC) have been suggested to arise from various developmental sources during embryogenesis, depending on the vascular bed. However, evidence also points to a common subpopulation of vascular progenitor cells predisposed to VSMC fate in the embryo. In the present study, we use binary transgenic reporter mice to identify a Tie1(+)CD31(dim)vascular endothelial (VE)-cadherin(-)CD45(-) precursor that gives rise to VSMC in vivo in all vascular beds examined. This precursor does not represent a mature endothelial cell, because a VE-cadherin promoter-driven reporter shows no expression in VSMC during murine development. Blockade of Notch signaling in the Tie1(+) precursor cell, but not the VE-cadherin(+) endothelial cell, decreases VSMC investment of developing arteries, leading to localized hemorrhage in the embryo at the time of vascular maturation. However, Notch signaling is not required in the Tie1(+) precursor after establishment of a stable artery. Thus, Notch activity is required in the differentiation of a Tie1(+) local precursor to VSMC in a spatiotemporal fashion across all vascular beds.
Noseda M, Mcsweeney SJ, Leja T, et al., 2012, Mutually exclusive expression of key cardiogenic transcription factors in single cardiac progenitor cells, 2nd Congress of the European-Society-of-Cardiology Council on Basic Cardiovascular Science - Frontiers in Cardiovascular Biology, Publisher: OXFORD UNIV PRESS, Pages: S16-S16, ISSN: 0008-6363
Noseda M, De Smith AJ, Leja T, et al., 2012, The Cdkn2a/2b tumour suppressor locus is a hot spot for genome instability in mouse cardiac progenitor cells, 2nd Congress of the European-Society-of-Cardiology Council on Basic Cardiovascular Science - Frontiers in Cardiovascular Biology, Publisher: OXFORD UNIV PRESS, Pages: S59-S59, ISSN: 0008-6363
Noseda M, Peterkin T, Simoes FC, et al., 2011, Cardiopoietic Factors Extracellular Signals for Cardiac Lineage Commitment, CIRCULATION RESEARCH, Vol: 108, Pages: 129-152, ISSN: 0009-7330
Lin S-C, Dolle P, Ryckebuesch L, et al., 2010, Endogenous retinoic acid regulates cardiac progenitor differentiation, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, Vol: 107, Pages: 9234-9239, ISSN: 0027-8424
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