DONA ORA
Pubblicazioni recenti - cardiac stem
-
Biomanufacturing of organ-specific tissues with high cellular density and embedded vascular channels.
Sci Adv2019 Sep;5(9):eaaw2459. doi: 10.1126/sciadv.aaw2459.
Skylar-Scott Mark A, Uzel Sebastien G M, Nam Lucy L, Ahrens John H, Truby Ryan L, Damaraju Sarita, Lewis Jennifer A,
Abstract
Engineering organ-specific tissues for therapeutic applications is a grand challenge, requiring the fabrication and maintenance of densely cellular constructs composed of ~10 cells/ml. Organ building blocks (OBBs) composed of patient-specific-induced pluripotent stem cell-derived organoids offer a pathway to achieving tissues with the requisite cellular density, microarchitecture, and function. However, to date, scant attention has been devoted to their assembly into 3D tissue constructs. Here, we report a biomanufacturing method for assembling hundreds of thousands of these OBBs into living matrices with high cellular density into which perfusable vascular channels are introduced via embedded three-dimensional bioprinting. The OBB matrices exhibit the desired self-healing, viscoplastic behavior required for sacrificial writing into functional tissue (SWIFT). As an exemplar, we created a perfusable cardiac tissue that fuses and beats synchronously over a 7-day period. Our SWIFT biomanufacturing method enables the rapid assembly of perfusable patient- and organ-specific tissues at therapeutic scales.
Guarda su PubMed
-
Moving forward on the pathway of cell-based therapies in ischemic heart disease and heart failure - time for new recommendations?
World J Stem Cells2019 Aug;11(8):445-451. doi: 10.4252/wjsc.v11.i8.445.
Micheu Miruna Mihaela,
Abstract
Although substantial advances have been made in treating ischemic heart disease and subsequent heart failure, the overall morbidity and mortality from these conditions remain high. Stem cell-based therapy has emerged as a promising approach for prompting cardiac rejuvenation. Various cell types have been tested in the clinical arena, proving consistent safety results. As for efficiency outcomes, contradictory findings have been reported, partly due to inconsistency in study protocols but also due to poor survival, engraftment and differentiation of transplanted cells in the hostile milieu of the ischemic host tissue. Studies have varied in terms of route of delivery, type and dose of implanted stem cells, patient selection and randomization, and assessment of therapeutic effect. Founded on the main achievements and challenges within almost 20 years of research, a number of official documents have been published by leading experts in the field. Core recommendations have focused on developing and optimizing effective strategies to enrich cell retention and their regenerative potential. Issued consensus and position papers have stemmed from an unmet need to provide a harmonized framework for future research, resulting in improved therapeutic application of cell-based therapies for cardiac regeneration and repair.
Guarda su PubMed
-
Detection of Na1.5 Conformational Change in Mammalian Cells Using the Noncanonical Amino Acid ANAP.
Biophys J2019 Aug;():. doi: S0006-3495(19)30745-3.
Shandell Mia A, Quejada Jose R, Yazawa Masayuki, Cornish Virginia W, Kass Robert S,
Abstract
Na1.5 inactivation is necessary for healthy conduction of the cardiac action potential. Genetic mutations of Na1.5 perturb inactivation and cause potentially fatal arrhythmias associated with long QT syndrome type 3. The exact structural dynamics of the inactivation complex is unknown. To sense inactivation gate conformational change in live mammalian cells, we incorporated the solvatochromic fluorescent noncanonical amino acid 3-((6-acetylnaphthalen-2-yl)amino)-2-aminopropanoic acid (ANAP) into single sites in the Na1.5 inactivation gate. ANAP was incorporated in full-length and C-terminally truncated Na1.5 channels using mammalian cell synthetase-tRNA technology. ANAP-incorporated channels were expressed in mammalian cells, and they exhibited pathophysiological function. A spectral imaging potassium depolarization assay was designed to detect ANAP emission shifts associated with Na1.5 conformational change. Site-specific intracellular ANAP incorporation affords live-cell imaging and detection of Na1.5 inactivation gate conformational change in mammalian cells.
Copyright © 2019 Biophysical Society. Published by Elsevier Inc. All rights reserved.
Guarda su PubMed
-
Comparative effects of parent and heated cinnamaldehyde on the function of human iPSC-derived cardiac myocytes.
Toxicol In Vitro2019 Sep;():104648. doi: S0887-2333(18)30817-8.
Nystoriak Matthew A, Kilfoil Peter J, Lorkiewicz Pawel K, Ramesh Bhargav, Kuehl Philip J, McDonald Jacob, Bhatnagar Aruni, Conklin Daniel J,
Abstract
Many e-cigarette products contain cinnamaldehyde as a primary constituent of cinnamon flavorings. When used as a food additive, cinnamaldehyde is generally regarded as safe for ingestion. However, little is known about the effects of cinnamaldehyde or its degradation products, generated after heating and inhalation, which may lead to elevated circulatory exposure to the heart. Hence, in this study, we tested the in vitro cardiac toxicity of cinnamaldehyde and its thermal degradation products generated by heating at low (200?±?50?°C) and high temperatures (700?±?50?°C) on the contractility, rhythmicity and electrical signaling properties of human induced pluripotent stem cell derived cardiac myocytes (hiPSC-CMs). Cellular impedance measurements on spontaneously beating hiPSC-CMs revealed that cinnamaldehyde significantly alters contraction-dependent signal amplitude, beating rate, and cell morphology. These effects were attenuated after cinnamaldehyde was subjected to heating at low or high temperatures. Current clamp analysis of hiPSC-CM action potentials (APs) showed only modest effects of acute application of 1-100??M cinnamaldehyde on resting membrane potential, while prolonged (~20?min) application of 100??M cinnamaldehyde resulted in progressive depolarization and loss of rhythmic AP spiking activity. Collectively, these results suggest that micromolar levels of cinnamaldehyde could alter cardiac excitability, in part by impairing the processes that regulate membrane potential and depolarization. Our results further suggest that heating cinnamaldehyde by itself does not directly lead to the formation of products with greater cardiotoxicity in vitro.
Copyright © 2018. Published by Elsevier Ltd.
Guarda su PubMed
-
Functional improvement and maturation of human cardiomyocytes derived from human pluripotent stem cells by barbaloin preconditioning.
Acta Biochim Biophys Sin (Shanghai)2019 Sep;():. doi: gmz090.
Yang Hui, Zhong Weiyi, Hamidi Mohammad Rafi, Zhou Gaojun, Liu Chen,
Abstract
The development of human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) is a significant advancement in our ability to obtain cardiomyocytes in vitro for regenerative therapies and drug discovery. However, hPSC-CMs obtained via existing protocols usually exhibit a markedly immature phenotype, compared with adult cardiomyocytes, thereby limiting their application. Here we report that barbaloin preconditioning dramatically improves the morphology, structure-related cardiac gene expression, calcium handling, and electrophysiological properties of hPSC-CMs, which means that barbaloin may have the potential to induce the maturation of hPSC-CMs, providing a novel strategy to generate more adult-like cardiomyocytes and promoting the application of hPSC-CMs in regenerative medicine, drug development, and disease modeling.
© The Author(s) 2018. Published by Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.
Guarda su PubMed
-
Single-Cell Delineation of Who's on First and Second Heart Fields During Development.
Circ Res2019 Aug;125(4):411-413. doi: 10.1161/CIRCRESAHA.119.315576.
Galdos Francisco X, Wu Sean M,
Guarda su PubMed
-
Native T1 time and extracellular volume fraction in differentiation of normal myocardium from non-ischemic dilated and hypertrophic cardiomyopathy myocardium: A systematic review and meta-analysis.
Int J Cardiol Heart Vasc2019 Dec;25():100422. doi: 10.1016/j.ijcha.2019.100422.
Minegishi Shintaro, Kato Shingo, Takase-Minegishi Kaoru, Horita Nobuyuki, Azushima Kengo, Wakui Hiromichi, Ishigami Tomoaki, Kosuge Masami, Kimura Kazuo, Tamura Kouichi,
Abstract
Background:
Both native T1 time and extracellular volume (ECV) fraction have been shown to be important measures for the detection of myocardial fibrosis. However, ECV determination requires the administration of an intravenous contrast agent, whereas native T1 mapping can be performed without a contrast agent.
Methods:
Here, we conducted a meta-analysis of myocardial native T1 data obtained for non-ischemic cardiomyopathy (NIC) patients and controls. A literature review included studies that applied T1 mapping using modified Look-Locker inversion recovery to measure myocardial fibrosis, and the results were validated by comparing datasets for dilated cardiomyopathy (DCM) or hypertrophic cardiomyopathy (HCM) patients and healthy controls (HCs).
Results:
We identified 16 eligible studies. Pooled mean differences (MDs) and 95% confidence intervals (CIs) were estimated as follows. Native T1 at 1.5-T, DCM vs. HC: MD?=?45.26 (95% CI: 30.92-59.59); HCM vs. HC: MD?=?47.09 (95% CI: 32.42-61.76). Native T1 at 3.0-T, DCM vs. HC: MD?=?82.52 (95% CI: 47.60-117.44); HCM vs. HC: MD?=?115.87 (95% CI: 50.71-181.04). ECV at 1.5-T, DCM vs. HC: MD?=?4.26 (95% CI: 3.06-5.46); HCM vs. HC: MD?=?1.49 (95% CI: -1.45-4.43). ECV at 3.0-T, DCM vs. HC: MD?=?8.40 (95% CI: 2.94-13.86); HCM vs. HC: MD?=?8.02 (95% CI: 5.45-1-0.59).
Conclusion:
In conclusion, native T1 values were significantly different between NIC patients and controls. Native T1 mapping may be a useful noninvasive method to detect diffuse myocardial fibrosis in NIC patients.
Guarda su PubMed
-
Assessment of temporal functional changes and miRNA profiling of human iPSC-derived cardiomyocytes.
Sci Rep2019 Sep;9(1):13188. doi: 10.1038/s41598-019-49653-5.
Kumar Naresh, Dougherty Julie A, Manring Heather R, Elmadbouh Ibrahim, Mergaye Muhamad, Czirok Andras, Greta Isai Dona, Belevych Andriy E, Yu Lianbo, Janssen Paul M L, Fadda Paolo, Gyorke Sandor, Ackermann Maegen A, Angelos Mark G, Khan Mahmood,
Abstract
Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) have been developed for cardiac cell transplantation studies more than a decade ago. In order to establish the hiPSC-CM-based platform as an autologous source for cardiac repair and drug toxicity, it is vital to understand the functionality of cardiomyocytes. Therefore, the goal of this study was to assess functional physiology, ultrastructural morphology, gene expression, and microRNA (miRNA) profiling at Wk-1, Wk-2 & Wk-4 in hiPSC-CMs in vitro. Functional assessment of hiPSC-CMs was determined by multielectrode array (MEA), Ca cycling and particle image velocimetry (PIV). Results demonstrated that Wk-4 cardiomyocytes showed enhanced synchronization and maturation as compared to Wk-1 & Wk-2. Furthermore, ultrastructural morphology of Wk-4 cardiomyocytes closely mimicked the non-failing (NF) adult human heart. Additionally, modulation of cardiac genes, cell cycle genes, and pluripotency markers were analyzed by real-time PCR and compared with NF human heart. Increasing expression of fatty acid oxidation enzymes at Wk-4 supported the switching to lipid metabolism. Differential regulation of 12 miRNAs was observed in Wk-1 vs Wk-4 cardiomyocytes. Overall, this study demonstrated that Wk-4 hiPSC-CMs showed improved functional, metabolic and ultrastructural maturation, which could play a crucial role in optimizing timing for cell transplantation studies and drug screening.
Guarda su PubMed
-
Cell cycle regulators control mesoderm specification in human pluripotent stem cells.
J Biol Chem2019 Sep;():. doi: jbc.RA119.008251.
Yiangou Loukia, Grandy Rodrigo A, Osnato Anna, Ortmann Daniel, Sinha Sanjay, Vallier Ludovic,
Abstract
The mesoderm is one of the three germ layers produced during gastrulation from which muscle, bones, kidneys, and the cardiovascular system originate. Understanding the mechanisms that control mesoderm specification could inform many applications, including the development of regenerative medicine therapies to manage diseases affecting these tissues. Here, we used human pluripotent stem cells (hPSCs) to investigate the role of cell cycle in mesoderm formation. To this end, using small molecules or conditional gene knockdown, we inhibited proteins controlling G1 and G2/M cell cycle phases during the differentiation of hPSCs into lateral plate, cardiac, and presomitic mesoderm. These loss-of-function experiments revealed that regulators of the G1 phase such as cyclin-dependent kinases (CDKs) and phosphorylation of retinoblastoma protein (pRb) are necessary for efficient mesoderm formation in a context-dependent manner. Further investigations disclosed that inhibition of the G2/M regulator CDK1 decreases bone morphogenetic protein (BMP) signaling activity specifically during lateral plate mesoderm formation while reducing fibroblast growth factor (FGF)/extracellular signaling-regulated kinase 1/2 (ERK1/2) activity in all mesoderm subtypes. Taken together, our findings reveal that cell cycle regulators direct mesoderm formation by controlling the activity of key developmental pathways.
Published under license by The American Society for Biochemistry and Molecular Biology, Inc.
Guarda su PubMed
-
Cardiac regenerative therapy: Many paths to repair.
Trends Cardiovasc Med2019 Sep;():. doi: S1050-1738(19)30127-6.
Gude Natalie A, Sussman Mark A,
Abstract
Cardiovascular disease remains the primary cause of death in the United States and in most nations worldwide, despite ongoing intensive efforts to promote cardiac health and treat heart failure. Replacing damaged myocardium represents perhaps the most promising treatment strategy, but also the most challenging given that the adult mammalian heart is notoriously resistant to endogenous repair. Cardiac regeneration following pathologic challenge would require proliferation of surviving tissue, expansion and differentiation of resident progenitors, or transdifferentiation of exogenously applied progenitor cells into functioning myocardium. Adult cardiomyocyte proliferation has been the focus of investigation for decades, recently enjoying a renaissance of interest as a therapeutic strategy for reversing cardiomyocyte loss due in large part to ongoing controversies and frustrations with myocardial cell therapy outcomes. The promise of cardiac cell therapy originated with reports of resident adult cardiac stem cells that could be isolated, expanded and reintroduced into damaged myocardium, producing beneficial effects in preclinical animal models. Despite modest functional improvements, Phase I clinical trials using autologous cardiac derived cells have proven safe and effective, setting the stage for an ongoing multi-center Phase II trial combining autologous cardiac stem cell types to enhance beneficial effects. This overview will examine the history of these two approaches for promoting cardiac repair and attempt to provide context for current and future directions in cardiac regenerative research.
Copyright © 2019 Elsevier Inc. All rights reserved.
Guarda su PubMed
-
Induction of Articular Chondrogenesis by Chitosan/Hyaluronic-Acid-Based Biomimetic Matrices Using Human Adipose-Derived Stem Cells.
Int J Mol Sci2019 Sep;20(18):. doi: E4487.
Huang Yijiang, Seitz Daniel, König Fabian, Müller Peter E, Jansson Volkmar, Klar Roland M,
Abstract
Cartilage repair using tissue engineering is the most advanced clinical application in regenerative medicine, yet available solutions remain unsuccessful in reconstructing native cartilage in its proprietary form and function. Previous investigations have suggested that the combination of specific bioactive elements combined with a natural polymer could generate carrier matrices that enhance activities of seeded stem cells and possibly induce the desired matrix formation. The present study sought to clarify this by assessing whether a chitosan-hyaluronic-acid-based biomimetic matrix in conjunction with adipose-derived stem cells could support articular hyaline cartilage formation in relation to a standard chitosan-based construct. By assessing cellular development, matrix formation, and key gene/protein expressions during in vitro cultivation utilizing quantitative gene and immunofluorescent assays, results showed that chitosan with hyaluronic acid provides a suitable environment that supports stem cell differentiation towards cartilage matrix producing chondrocytes. However, on the molecular gene expression level, it has become apparent that, without combinations of morphogens, in the chondrogenic medium, hyaluronic acid with chitosan has a very limited capacity to stimulate and maintain stem cells in an articular chondrogenic state, suggesting that cocktails of various growth factors are one of the key features to regenerate articular cartilage, clinically.
Guarda su PubMed
-
Leducq Transatlantic Network of Excellence to Cure Phospholamban-Induced Cardiomyopathy (CURE-PLaN).
Circ Res2019 Sep;125(7):720-724. doi: 10.1161/CIRCRESAHA.119.315077.
Doevendans Pieter A, Glijnis Pieter C, Kranias Evangelia G,
Abstract
The deletion of Arginine 14 of the phosholamban gene (PLN p.R14del) is associated with the pathogenesis of an inherited form of cardiomyopathy with prominent arrhythmias. Patients carrying the PLN R14del mutation are at risk of developing dilated cardiomyopathy or arrhythmogenic right ventricular cardiomyopathy. Although the genetic etiology is well defined, the molecular mechanism underlying the pathogenesis of PLN R14del-cardiomyopathy is unknown. Our CURE PLaN network, funded by the Foundation Leducq, will bring together leading scientists, clinicians, and patients to elucidate the genotype-phenotype relationships in R14del cardiomyopathy with the ultimate goal of developing innovative disease-specific therapeutic modalities. With the generous support of the Leducq Foundation, our Transatlantic Network of Excellence consortium to cure Phospholamban (PLN)-induced cardiomyopathy (CURE-PLaN) unites 6 leading centers to address the current challenges associated with arrhythmogenic right ventricular cardiomyopathy/dilated cardiomyopathy (DCM) with an initial focus on PLN and development of effective treatments. The Network is led by Evangelia (Litsa) Kranias (University of Cincinnati) in the United States and Pieter A. Doevendans (Netherlands Heart Institute/UMC Utrecht NL) in Europe. The other US project leaders are Kevin Costa (Cardiovascular Research Center, Icahn School of Medicine at Mount Sinai, New York) and Mark Mercola and Ioannis Karakikes (Stanford University), who are focusing on induced pluripotent stem cell (iPSC)-based disease models, tissue engineering, gene therapy, and drug discovery. On the European side, the project leaders are Despina Sanoudou (Biomedical Research Foundation of the Academy of Athens) analyzing the PLN interactome and Stephan Lehnart (University of Gottingen) addressing the subcellular and disrupted protein interactions affected in PLN-mutant cardiomyocytes. Other key members within the Netherlands Heart Institute are Peter van Tintelen on PLN genetics, Folkert Asselbergs on epigenetics and Rudolf de Boer on clinical trials. We are also privileged to get support from Arthur Wilde (University of Amsterdam), Sakthivel Sadayappan (University of Cincinnati), and Roger Hajjar (Phospholamban Foundation), who have had a long-standing interest in cardiac physiology and pathophysiology with emphasis on underlying pathways and potential therapeutic targets. The consortium is also fortunate to embrace a patient advocate, Pieter Glijnis, incorporating the voice of the patients to research in every step. Our goal is to build and share a platform of patient data coupled with in vitro and in vivo models to promote scientific discovery and advance novel treatments. Phospholamban is a small phosphoprotein in the cardiac sarcoplasmic reticulum, and it is the major regulator of SERCA2a activity and calcium (Ca)-cycling. Chronic inhibition of SERCA2a by PLN has been implicated in the aberrant Ca-cycling of failing hearts. Studies in HF models have shown that decreasing PLN activity may rescue cardiac remodeling and dysfunction. Several human PLN mutations, leading to inhibition of Ca-uptake into the sarcoplasmic reticulum, are linked to inherited DCM.
Guarda su PubMed
-
Novel synonymous and missense variants in FGFR1 causing Hartsfield syndrome.
Am J Med Genet A2019 Sep;():. doi: 10.1002/ajmg.a.61354.
Courage Carolina, Jackson Christopher B, Owczarek-Lipska Marta, Jamsheer Aleksander, Sowi?ska-Seidler Anna, Piotrowicz Ma?gorzata, Jakubowski Lucjusz, Dallčves Fanny, Riesch Erik, Neidhardt John, Lemke Johannes R,
Abstract
Hartsfield syndrome is a rare clinical entity characterized by holoprosencephaly and ectrodactyly with the variable feature of cleft lip/palate. In addition to these symptoms patients with Hartsfield syndrome can show developmental delay of variable severity, isolated hypogonadotropic hypogonadism, central diabetes insipidus, vertebral anomalies, eye anomalies, and cardiac malformations. Pathogenic variants in FGFR1 have been described to cause phenotypically different FGFR1-related disorders such as Hartsfield syndrome, hypogonadotropic hypogonadism with or without anosmia, Jackson-Weiss syndrome, osteoglophonic dysplasia, Pfeiffer syndrome, and trigonocephaly Type 1. Here, we report three patients with Hartsfield syndrome from two unrelated families. Exome sequencing revealed two siblings harboring a novel de novo heterozygous synonymous variant c.1029G>A, p.Ala343Ala causing a cryptic splice donor site in exon 8 of FGFR1 likely due to gonadal mosaicism in one parent. The third case was a sporadic patient with a novel de novo heterozygous missense variant c.1868A>G, p.(Asp623Gly).
© 2019 Wiley Periodicals, Inc.
Guarda su PubMed
-
Mesenchymal stem cells-derived extracellular vesicles in acute respiratory distress syndrome: a review of current literature and potential future treatment options.
Clin Transl Med2019 Sep;8(1):25. doi: 10.1186/s40169-019-0242-9.
Shah Trushil G, Predescu Dan, Predescu Sanda,
Abstract
Acute respiratory distress syndrome (ARDS) is a life-threatening inflammatory lung condition associated with significant morbidity and mortality. Unfortunately, the current treatment for this disease is mainly supportive. Mesenchymal stem cells (MSCs) due to their immunomodulatory properties are increasingly being studied for the treatment of ARDS and have shown promise in multiple animal studies. The therapeutic effects of MSCs are exerted in part in a paracrine manner by releasing extracellular vesicles (EVs), rather than local engraftment. MSC-derived EVs are emerging as potential alternatives to MSC therapy in ARDS. In this review, we will introduce EVs and briefly discuss current data on EVs and MSCs in ARDS. We will discuss current literature on the role of MSC-derived EVs in pathogenesis and treatment of ARDS and their potential as a treatment strategy in the future.
Guarda su PubMed
-
A middle Cambrian arthropod with chelicerae and proto-book gills.
Nature2019 Sep;():. doi: 10.1038/s41586-019-1525-4.
Aria Cédric, Caron Jean-Bernard,
Abstract
The chelicerates are a ubiquitous and speciose group of animals that has a considerable ecological effect on modern terrestrial ecosystems-notably as predators of insects and also, for instance, as decomposers. The fossil record shows that chelicerates diversified early in the marine ecosystems of the Palaeozoic era, by at least the Ordovician period. However, the timing of chelicerate origins and the type of body plan that characterized the earliest members of this group have remained controversial. Although megacheirans have previously been interpreted as chelicerate-like, and habeliidans (including Sanctacaris) have been suggested to belong to their immediate stem lineage, evidence for the specialized feeding appendages (chelicerae) that are diagnostic of the chelicerates has been lacking. Here we use exceptionally well-preserved and abundant fossil material from the middle Cambrian Burgess Shale (Marble Canyon, British Columbia, Canada) to show that Mollisonia plenovenatrix sp. nov. possessed robust but short chelicerae that were placed very anteriorly, between the eyes. This suggests that chelicerae evolved a specialized feeding function early on, possibly as a modification of short antennules. The head also encompasses a pair of large compound eyes, followed by three pairs of long, uniramous walking legs and three pairs of stout, gnathobasic masticatory appendages; this configuration links habeliidans with euchelicerates ('true' chelicerates, excluding the sea spiders). The trunk ends in a four-segmented pygidium and bears eleven pairs of identical limbs, each of which is composed of three broad lamellate exopod flaps, and endopods are either reduced or absent. These overlapping exopod flaps resemble euchelicerate book gills, although they lack the diagnostic operculum. In addition, the eyes of M. plenovenatrix were innervated by three optic neuropils, which strengthens the view that a complex malacostracan-like visual system might have been plesiomorphic for all crown euarthropods. These fossils thus show that chelicerates arose alongside mandibulates as benthic micropredators, at the heart of the Cambrian explosion.
Guarda su PubMed
-
?Klotho Regulates Age-Associated Vascular Calcification and Lifespan in Zebrafish.
Cell Rep2019 Sep;28(11):2767-2776.e5. doi: S2211-1247(19)31046-0.
Singh Ajeet Pratap, Sosa Maria X, Fang Jian, Shanmukhappa Shiva Kumar, Hubaud Alexis, Fawcett Caroline H, Molind Gregory J, Tsai Tingwei, Capodieci Paola, Wetzel Kristie, Sanchez Ellen, Wang Guangliang, Coble Matthew, Tang Wenlong, Cadena Samuel M, Fishman Mark C, Glass David J,
Abstract
The hormone ?Klotho regulates lifespan in mice, as knockouts die early of what appears to be accelerated aging due to hyperphosphatemia and soft tissue calcification. In contrast, the overexpression of ?Klotho increases lifespan. Given the severe mouse phenotype, we generated zebrafish mutants for ?klotho as well as its binding partner fibroblast growth factor-23 (fgf23). Both mutations cause shortened lifespan in zebrafish, with abrupt onset of behavioral and degenerative physical changes at around 5 months of age. There is a calcification of vessels throughout the body, most dramatically in the outflow tract of the heart, the bulbus arteriosus (BA). This calcification is associated with an ectopic activation of osteoclast differentiation pathways. These findings suggest that the gradual loss of ?Klotho found in normal aging might give rise to ectopic calcification.
Copyright © 2019 Novartis Institutes for Biomedical Research. Published by Elsevier Inc. All rights reserved.
Guarda su PubMed
-
Discovery of Retinoic Acid Receptor Agonists as Proliferators of Cardiac Progenitor Cells Through a Phenotypic Screening Approach.
Stem Cells Transl Med2019 Sep;():. doi: 10.1002/sctm.19-0069.
Drowley Lauren, McPheat Jane, Nordqvist Anneli, Peel Samantha, Karlsson Ulla, Martinsson Sofia, Müllers Erik, Dellsén Anita, Knight Sinead, Barrett Ian, Sánchez José, Magnusson Björn, Greber Boris, Wang Qing-Dong, Plowright Alleyn T,
Abstract
Identification of small molecules with the potential to selectively proliferate cardiac progenitor cells (CPCs) will aid our understanding of the signaling pathways and mechanisms involved and could ultimately provide tools for regenerative therapies for the treatment of post-MI cardiac dysfunction. We have used an in vitro human induced pluripotent stem cell-derived CPC model to screen a 10,000-compound library containing molecules representing different target classes and compounds reported to modulate the phenotype of stem or primary cells. The primary readout of this phenotypic screen was proliferation as measured by nuclear count. We identified retinoic acid receptor (RAR) agonists as potent proliferators of CPCs. The CPCs retained their progenitor phenotype following proliferation and the identified RAR agonists did not proliferate human cardiac fibroblasts, the major cell type in the heart. In addition, the RAR agonists were able to proliferate an independent source of CPCs, HuES6. The RAR agonists had a time-of-differentiation-dependent effect on the HuES6-derived CPCs. At 4?days of differentiation, treatment with retinoic acid induced differentiation of the CPCs to atrial cells. However, after 5?days of differentiation treatment with RAR agonists led to an inhibition of terminal differentiation to cardiomyocytes and enhanced the proliferation of the cells. RAR agonists, at least transiently, enhance the proliferation of human CPCs, at the expense of terminal cardiac differentiation. How this mechanism translates in vivo to activate endogenous CPCs and whether enhancing proliferation of these rare progenitor cells is sufficient to enhance cardiac repair remains to be investigated. Stem Cells Translational Medicine2019.
© 2019 The Authors. Stem Cells Translational Medicine published by Wiley Periodicals, Inc. on behalf of AlphaMed Press.
Guarda su PubMed
-
Optimizing Modified mRNA Synthesis Protocol for Heart Gene Therapy.
Mol Ther Methods Clin Dev2019 Sep;14():300-305. doi: 10.1016/j.omtm.2019.07.006.
Hadas Yoav, Sultana Nishat, Youssef Elias, Sharkar Mohammad Tofael Kabir, Kaur Keerat, Chepurko Elena, Zangi Lior,
Abstract
Synthetic modified RNA (modRNA) is a novel vector for gene transfer to the heart and other organs. modRNA can mediate strong, transient protein expression with minimal induction of the innate immune response and risk for genome integration. modRNA is already being used in several human clinical trials, and its use in basic and translational science is growing. Due to the complexity of preparing modRNA and the high cost of its reagents, there is a need for an improved, cost-efficient protocol to make modRNA. Here we show that changing the ratio between anti-reverse cap analog (ARCA) and N1-methyl-pseudouridine (N1m?), favoring ARCA over N1m?, significantly increases the yield per reaction, improves modRNA translation, and reduces its immunogenicity . This protocol will make modRNA preparation more accessible and financially affordable for basic and translational research.
Guarda su PubMed
-
Downregulation of miR-146a Contributes to Cardiac Dysfunction Induced by the Tyrosine Kinase Inhibitor Sunitinib.
Front Pharmacol2019 ;10():914. doi: 10.3389/fphar.2019.00914.
Shen Li, Li Congxin, Zhang Hua, Qiu Suhua, Fu Tian, Xu Yanfang,
Abstract
The main adverse effect of tyrosine kinase inhibitors, such as sunitinib, is cardiac contractile dysfunction; however, the molecular mechanisms of this effect remain largely obscure. MicroRNAs (miRNAs) are key regulatory factors in both cardiovascular diseases and the tyrosine kinase pathway. Therefore, we analyzed the differential expression of miRNAs in the myocardium in mice after exposure to sunitinib using miRNA microarray. A significant downregulation of miR-146a was observed in the myocardium of sunitinib-treated mice, along with a 20% decrease in left ventricle ejection fraction (LVEF). The downregulation of miR-146a was further validated by RT-qPCR. Among the potential targets of miR-146a, we focused on and , which are closely related to cardiac contractile dysfunction. Results of luciferase reporter assay confirmed that miR-146a directly targeted the 3' untranslated region of and . Significant upregulation of PLN and ANK2 at the mRNA and protein levels was observed in the myocardium of sunitinib-treated mice. Cardiac-specific overexpression of miR-146a prevented the deteriorate effect of SNT on calcium transients, thereby alleviating the decreased contractility of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). SiRNA knockdown of PLN or ANK2 prevented sunitinib-induced suppression of contractility in hiPSC-CMs. Therefore, our and results showed that sunitinib downregulated miR-146a, which contributes to cardiac contractile dysfunction by regulating the downstream targets PLN and ANK2, and that upregulation of miR-146a alleviated the inhibitory effect of SNT on cardiac contractility. Thus, miR-146a could be a useful protective agent against sunitinib-induced cardiac dysfunction.
Guarda su PubMed
-
Effects of cigarette smoke components on myocardial differentiation of mouse embryonic stem cells.
Environ Toxicol2019 Sep;():. doi: 10.1002/tox.22843.
Kim Cho-Won, Go Ryeo-Eun, Ko Eul Bee, Jeung Eui-Bae, Kim Min-Seok, Choi Kyung-Chul,
Abstract
The heart is the first organ formed in the developing fetus, and abnormal development of the heart is a major cause of fetal death. The adverse effects of cigarette smoke on the heart have been well established, but it is not well understood how cigarette smoke components regulate signaling molecules and cardiac specific functions during the early differentiation stage of the embryonic heart. In this study, we identified changes in the size of mouse embryoid bodies (mEBs) in response to treatment with cigarette smoke extract (CSE) via regulation of HDAC2, p53, p21, and cyclin D1 protein expression, which are cardiac differentiation and cell-cycle markers, respectively. In addition, exposure of mouse embryonic stem cells (mESCs) to cigarette smoke components inhibited myocardial differentiation and development through the expression of HDAC1, HDAC2, GATA4, NKX2-5, TBX5, HAND1, and Troponin I. Long-term exposure studies showed that CSE and nicotine may delay the development of mouse cardiomyocytes from mESCs and inhibit the contractibility, which is a fundamental function of the heart. Taken together, these findings suggest that cigarette smoke components, including nicotine, may affect abnormal myocardial differentiation and development.
© 2019 Wiley Periodicals, Inc.
Guarda su PubMed
