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Article ; Online: Nr2f1a maintains atrial

Martin, Kendall E / Ravisankar, Padmapriyadarshini / Beerens, Manu / MacRae, Calum A / Waxman, Joshua S

2023 Volume 12

Abstract: Maintenance of cardiomyocyte identity is vital for normal heart development and function. However, our understanding of cardiomyocyte plasticity remains incomplete. Here, we show that sustained expression of the zebrafish transcription factor Nr2f1a ... ...

Abstract	Maintenance of cardiomyocyte identity is vital for normal heart development and function. However, our understanding of cardiomyocyte plasticity remains incomplete. Here, we show that sustained expression of the zebrafish transcription factor Nr2f1a prevents the progressive acquisition of ventricular cardiomyocyte (VC) and pacemaker cardiomyocyte (PC) identities within distinct regions of the atrium. Transcriptomic analysis of flow-sorted atrial cardiomyocytes (ACs) from
MeSH term(s)	Animals ; Atrial Fibrillation ; Gene Expression Regulation, Developmental ; Homeobox Protein Nkx-2.5/genetics ; Homeobox Protein Nkx-2.5/metabolism ; Homeodomain Proteins/metabolism ; Myocytes, Cardiac/metabolism ; Transcription Factors/genetics ; Transcription Factors/metabolism ; Zebrafish/genetics ; Zebrafish Proteins/genetics ; Zebrafish Proteins/metabolism
Chemical Substances	Homeobox Protein Nkx-2.5 ; Homeodomain Proteins ; Transcription Factors ; Zebrafish Proteins ; nr2f1a protein, zebrafish
Language	English
Publishing date	2023-05-15
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2687154-3
ISSN	2050-084X ; 2050-084X
ISSN (online)	2050-084X
ISSN	2050-084X
DOI	10.7554/eLife.77408
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Article ; Online: Repetitive Antigen Responses of LDL-Reactive CD4+ T Cells Induce Tr1 Cell-Mediated Immune Tolerance.

Mailer, Reiner K / Konrath, Sandra / Zhan, Lydia / Thode, Hanna / Beerens, Manu / Frye, Maike / Ketelhuth, Daniel F J / Renné, Thomas / Hansson, Göran K

Arteriosclerosis, thrombosis, and vascular biology

2023 Volume 43, Issue 8, Page(s) 1510–1523

Abstract: Background: Inflammation triggered by the deposition of LDL (low-density lipoprotein) in the arterial wall leads to the development of atherosclerosis. Regulatory T (Treg) cells inhibit vascular inflammation through the induction of immune tolerance ... ...

Abstract	Background: Inflammation triggered by the deposition of LDL (low-density lipoprotein) in the arterial wall leads to the development of atherosclerosis. Regulatory T (Treg) cells inhibit vascular inflammation through the induction of immune tolerance toward LDL-related antigens. However, tolerogenic mechanisms that promote the generation of LDL-specific Treg cells in vivo remain unclear. Methods: We identified LDL-specific T cells by activation-induced marker expression and analyzed expression profiles and suppressive functions of TCR (T-cell antigen receptor)-transgenic T cells upon repetitive transfer into antigen-transgenic mice via flow cytometry. Results: We investigated the naturally occurring Treg-cell response against human LDL in standard chow diet-fed mice that are transgenic for human ApoB100 (apolipoprotein B100). We found that IL (interleukin)-10 expression in LDL-specific T cells from spleen increases with age, albeit LDL-specific populations do not enlarge in older mice. To investigate the generation of IL-10-producing LDL-specific T cells, we transferred naive CD4+ T cells recognizing human ApoB100 from TCR-transgenic mice into human ApoB100-transgenic mice. Adoptive transfer of human ApoB100-specific T cells induced immune tolerance in recipient mice and effectively inhibited activation of subsequently transferred naive T cells of the same specificity in vivo. Moreover, repetitive transfers increased the population of Treg type 1 cells that suppress ApoB100-specific responses via IL-10. In a translational approach, LDL-specific Treg type 1 cells from blood of healthy donors suppressed the activation of monocytic THP-1 cells in an IL-10-dependent manner. Conclusions: We show that repetitive transfer of naive ApoB100-specific T cells and recurrent LDL-specific T-cell stimulation induces Treg type 1 cell-mediated immune tolerance against LDL in vivo. Our results provide insight into the generation of autoantigen-specific anti-inflammatory T cells under tolerogenic conditions.
MeSH term(s)	Mice ; Humans ; Animals ; T-Lymphocytes, Regulatory ; CD4-Positive T-Lymphocytes ; Interleukin-10/genetics ; Mice, Transgenic ; Immune Tolerance ; Receptors, Antigen, T-Cell/metabolism ; Inflammation/metabolism
Chemical Substances	Interleukin-10 (130068-27-8) ; Receptors, Antigen, T-Cell
Language	English
Publishing date	2023-06-01
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1221433-4
ISSN	1524-4636 ; 1079-5642
ISSN (online)	1524-4636
ISSN	1079-5642
DOI	10.1161/ATVBAHA.123.319135
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Zs.A 1705: Show issues

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Article ; Online: Targeting NETs using dual-active DNase1 variants.

Englert, Hanna / Göbel, Josephine / Khong, Danika / Omidi, Maryam / Wolska, Nina / Konrath, Sandra / Frye, Maike / Mailer, Reiner K / Beerens, Manu / Gerwers, Julian C / Preston, Roger J S / Odeberg, Jacob / Butler, Lynn M / Maas, Coen / Stavrou, Evi X / Fuchs, Tobias A / Renné, Thomas

Frontiers in immunology

2023 Volume 14, Page(s) 1181761

Abstract: Background: Neutrophil Extracellular Traps (NETs) are key mediators of immunothrombotic mechanisms and defective clearance of NETs from the circulation underlies an array of thrombotic, inflammatory, infectious, and autoimmune diseases. Efficient NET ... ...

Abstract	Background: Neutrophil Extracellular Traps (NETs) are key mediators of immunothrombotic mechanisms and defective clearance of NETs from the circulation underlies an array of thrombotic, inflammatory, infectious, and autoimmune diseases. Efficient NET degradation depends on the combined activity of two distinct DNases, DNase1 and DNase1-like 3 (DNase1L3) that preferentially digest double-stranded DNA (dsDNA) and chromatin, respectively. Methods: Here, we engineered a dual-active DNase with combined DNase1 and DNase1L3 activities and characterized the enzyme for its NET degrading potential in vitro. Furthermore, we produced a mouse model with transgenic expression of the dual-active DNase and analyzed body fluids of these animals for DNase1 and DNase 1L3 activities. We systematically substituted 20 amino acid stretches in DNase1 that were not conserved among DNase1 and DNase1L3 with homologous DNase1L3 sequences. Results: We found that the ability of DNase1L3 to degrade chromatin is embedded into three discrete areas of the enzyme's core body, not the C-terminal domain as suggested by the state-of-the-art. Further, combined transfer of the aforementioned areas of DNase1L3 to DNase1 generated a dual-active DNase1 enzyme with additional chromatin degrading activity. The dual-active DNase1 mutant was superior to native DNase1 and DNase1L3 in degrading dsDNA and chromatin, respectively. Transgenic expression of the dual-active DNase1 mutant in hepatocytes of mice lacking endogenous DNases revealed that the engineered enzyme was stable in the circulation, released into serum and filtered to the bile but not into the urine. Conclusion: Therefore, the dual-active DNase1 mutant is a promising tool for neutralization of DNA and NETs with potential therapeutic applications for interference with thromboinflammatory disease states.
MeSH term(s)	Mice ; Animals ; Endodeoxyribonucleases/genetics ; Extracellular Traps/metabolism ; Deoxyribonuclease I/genetics ; Deoxyribonuclease I/metabolism ; Chromatin ; DNA/metabolism ; Deoxyribonucleases/genetics
Chemical Substances	Endodeoxyribonucleases (EC 3.1.-) ; Deoxyribonuclease I (EC 3.1.21.1) ; Chromatin ; DNA (9007-49-2) ; Deoxyribonucleases (EC 3.1.-)
Language	English
Publishing date	2023-05-23
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2023.1181761
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Article ; Online: The phosphodiesterase 2A controls lymphatic junctional maturation via cGMP-dependent notch signaling.

Carlantoni, Claudia / Liekfeld, Leon M H / Hemkemeyer, Sandra A / Schreier, Danny / Saygi, Ceren / Kurelic, Roberta / Cardarelli, Silvia / Kalucka, Joanna / Schulte, Christian / Beerens, Manu / Mailer, Reiner K / Schäffer, Tilman E / Naro, Fabio / Pellegrini, Manuela / Nikolaev, Viacheslav O / Renné, Thomas / Frye, Maike

Developmental cell

2023 Volume 59, Issue 3, Page(s) 308–325.e11

Abstract: The molecular mechanisms by which lymphatic vessels induce cell contact inhibition are not understood. Here, we identify the cGMP-dependent phosphodiesterase 2A (PDE2A) as a selective regulator of lymphatic but not of blood endothelial contact inhibition. ...

Abstract	The molecular mechanisms by which lymphatic vessels induce cell contact inhibition are not understood. Here, we identify the cGMP-dependent phosphodiesterase 2A (PDE2A) as a selective regulator of lymphatic but not of blood endothelial contact inhibition. Conditional deletion of Pde2a in mouse embryos reveals severe lymphatic dysplasia, whereas blood vessel architecture remains unaltered. In the absence of PDE2A, human lymphatic endothelial cells fail to induce mature junctions and cell cycle arrest, whereas cGMP levels, but not cAMP levels, are increased. Loss of PDE2A-mediated cGMP hydrolysis leads to the activation of p38 signaling and downregulation of NOTCH signaling. However, DLL4-induced NOTCH activation restores junctional maturation and contact inhibition in PDE2A-deficient human lymphatic endothelial cells. In postnatal mouse mesenteries, PDE2A is specifically enriched in collecting lymphatic valves, and loss of Pde2a results in the formation of abnormal valves. Our data demonstrate that PDE2A selectively finetunes a crosstalk of cGMP, p38, and NOTCH signaling during lymphatic vessel maturation.
MeSH term(s)	Animals ; Humans ; Mice ; Cyclic Nucleotide Phosphodiesterases, Type 2/genetics ; Cyclic Nucleotide Phosphodiesterases, Type 2/metabolism ; Down-Regulation ; Endothelial Cells/metabolism ; Lymphatic Vessels/metabolism ; Signal Transduction
Chemical Substances	Cyclic Nucleotide Phosphodiesterases, Type 2 (EC 3.1.4.17) ; Pde2a protein, mouse (EC 3.1.4.17) ; PDE2A protein, human (EC 3.1.4.17)
Language	English
Publishing date	2023-12-29
Publishing country	United States
Document type	Journal Article
ZDB-ID	2054967-2
ISSN	1878-1551 ; 1534-5807
ISSN (online)	1878-1551
ISSN	1534-5807
DOI	10.1016/j.devcel.2023.12.002
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Zs.A 5742: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
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Article: Intrinsic coagulation pathway-mediated thrombin generation in mouse whole blood.

Konrath, Sandra / Mailer, Reiner K / Beerens, Manu / Englert, Hanna / Frye, Maike / Kuta, Piotr / Preston, Roger J S / Maas, Coen / Butler, Lynn M / Roest, Mark / de Laat, Bas / Renné, Thomas

Frontiers in cardiovascular medicine

2022 Volume 9, Page(s) 1008410

Abstract: Calibrated Automated Thrombography (CAT) is a versatile and sensitive method for analyzing coagulation reactions culminating in thrombin generation (TG). Here, we present a CAT method for analyzing TG in murine whole blood by adapting the CAT assay used ... ...

Abstract	Calibrated Automated Thrombography (CAT) is a versatile and sensitive method for analyzing coagulation reactions culminating in thrombin generation (TG). Here, we present a CAT method for analyzing TG in murine whole blood by adapting the CAT assay used for measuring TG in human plasma. The diagnostically used artificial and physiologic factor XII (FXII) contact activators kaolin, ellagic acid and polyphosphate (polyP) stimulated TG in murine blood in a dose-dependent manner resulting in a gradual increase in endogenous thrombin potential and peak thrombin, with shortened lag times and times to peak. The activated FXII inhibitor rHA-Infestin-4 and direct oral anticoagulants (DOACs) interfered with TG triggered by kaolin, ellagic acid and polyP and TG was completely attenuated in blood of FXII- (
Language	English
Publishing date	2022-11-28
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2781496-8
ISSN	2297-055X
ISSN	2297-055X
DOI	10.3389/fcvm.2022.1008410
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Article ; Online: PIEZO1 mediates a mechanothrombotic pathway in diabetes.

Zhu, Wandi / Guo, Shihui / Homilius, Max / Nsubuga, Cissy / Wright, Shane H / Quan, Dajun / Kc, Ashmita / Eddy, Samuel S / Victorio, Rachelle A / Beerens, Manu / Flaumenhaft, Robert / Deo, Rahul C / MacRae, Calum A

Science translational medicine

2022 Volume 14, Issue 626, Page(s) eabk1707

Abstract: Thrombosis is the leading complication of common human disorders including diabetes, coronary heart disease, and infection and remains a global health burden. Current anticoagulant therapies that target the general clotting cascade are associated with ... ...

Abstract	Thrombosis is the leading complication of common human disorders including diabetes, coronary heart disease, and infection and remains a global health burden. Current anticoagulant therapies that target the general clotting cascade are associated with unpredictable adverse bleeding effects, because understanding of hemostasis remains incomplete. Here, using perturbational screening of patient peripheral blood samples for latent phenotypes, we identified dysregulation of the major mechanosensory ion channel Piezo1 in multiple blood lineages in patients with type 2 diabetes mellitus (T2DM). Hyperglycemia activated
MeSH term(s)	Animals ; Diabetes Mellitus, Type 2 ; Humans ; Hyperglycemia/complications ; Ion Channels/metabolism ; Mechanotransduction, Cellular ; Thrombosis ; Zebrafish/metabolism ; Zebrafish Proteins/metabolism
Chemical Substances	Ion Channels ; PIEZO1 protein, human ; Piezo1 protein, zebrafish ; Zebrafish Proteins
Language	English
Publishing date	2022-01-05
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2518854-9
ISSN	1946-6242 ; 1946-6234
ISSN (online)	1946-6242
ISSN	1946-6234
DOI	10.1126/scitranslmed.abk1707
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Article ; Online: Stx4 is required to regulate cardiomyocyte Ca

Perl, Eliyahu / Ravisankar, Padmapriyadarshini / Beerens, Manu E / Mulahasanovic, Lejla / Smallwood, Kelly / Sasso, Marion Bermúdez / Wenzel, Carina / Ryan, Thomas D / Komár, Matej / Bove, Kevin E / MacRae, Calum A / Weaver, K Nicole / Prada, Carlos E / Waxman, Joshua S

HGG advances

2022 Volume 3, Issue 3, Page(s) 100115

Abstract: Requirements for vesicle fusion within the heart remain poorly understood, despite the multitude of processes that necessitate proper intracellular trafficking within cardiomyocytes. Here, we show that Syntaxin 4 (STX4), a target- ... ...

Abstract	Requirements for vesicle fusion within the heart remain poorly understood, despite the multitude of processes that necessitate proper intracellular trafficking within cardiomyocytes. Here, we show that Syntaxin 4 (STX4), a target-Soluble
Language	English
Publishing date	2022-04-27
Publishing country	United States
Document type	Journal Article
ISSN	2666-2477
ISSN (online)	2666-2477
DOI	10.1016/j.xhgg.2022.100115
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Article ; Online: RBPMS2 Is a Myocardial-Enriched Splicing Regulator Required for Cardiac Function.

Akerberg, Alexander A / Trembley, Michael / Butty, Vincent / Schwertner, Asya / Zhao, Long / Beerens, Manu / Liu, Xujie / Mahamdeh, Mohammed / Yuan, Shiaulou / Boyer, Laurie / MacRae, Calum / Nguyen, Christopher / Pu, William T / Burns, Caroline E / Burns, C Geoffrey

Circulation research

2022 Volume 131, Issue 12, Page(s) 980–1000

Abstract: Background: RBPs (RNA-binding proteins) perform indispensable functions in the post-transcriptional regulation of gene expression. Numerous RBPs have been implicated in cardiac development or physiology based on gene knockout studies and the ... ...

Abstract	Background: RBPs (RNA-binding proteins) perform indispensable functions in the post-transcriptional regulation of gene expression. Numerous RBPs have been implicated in cardiac development or physiology based on gene knockout studies and the identification of pathogenic RBP gene mutations in monogenic heart disorders. The discovery and characterization of additional RBPs performing indispensable functions in the heart will advance basic and translational cardiovascular research. Methods: We performed a differential expression screen in zebrafish embryos to identify genes enriched in Results: We identified 1848 genes enriched in the Conclusions: Our study identifies
MeSH term(s)	Animals ; Humans ; Calcium/metabolism ; Myocardium/metabolism ; Myocytes, Cardiac/metabolism ; Repressor Proteins/metabolism ; RNA Splicing Factors/metabolism ; RNA-Binding Proteins/genetics ; RNA-Binding Proteins/metabolism ; Zebrafish/genetics ; Zebrafish/metabolism ; Zebrafish Proteins/genetics ; Zebrafish Proteins/metabolism
Chemical Substances	Calcium (SY7Q814VUP) ; Repressor Proteins ; RNA Splicing Factors ; RNA-Binding Proteins ; Zebrafish Proteins ; RBPMS2 protein, human
Language	English
Publishing date	2022-11-11
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S. ; Research Support, Non-U.S. Gov't
ZDB-ID	80100-8
ISSN	1524-4571 ; 0009-7330 ; 0931-6876
ISSN (online)	1524-4571
ISSN	0009-7330 ; 0931-6876
DOI	10.1161/CIRCRESAHA.122.321728
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Ui IV Zs.70: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 2021: Bestellungen von Artikeln über das Online-Bestellformular ab Jg. 2022: Lesesaal (EG)
Ui IV Zs.60: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 2021: Bestellungen von Artikeln über das Online-Bestellformular ab Jg. 2022: Lesesaal (EG)

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Article ; Online: MIC-Drop: A platform for large-scale in vivo CRISPR screens.

Parvez, Saba / Herdman, Chelsea / Beerens, Manu / Chakraborti, Korak / Harmer, Zachary P / Yeh, Jing-Ruey J / MacRae, Calum A / Yost, H Joseph / Peterson, Randall T

Science (New York, N.Y.)

2021 Volume 373, Issue 6559, Page(s) 1146–1151

Abstract: CRISPR-Cas9 can be scaled up for large-scale screens in cultured cells, but CRISPR screens in animals have been challenging because generating, validating, and keeping track of large numbers of mutant animals is prohibitive. Here, we introduce ... ...

Abstract	CRISPR-Cas9 can be scaled up for large-scale screens in cultured cells, but CRISPR screens in animals have been challenging because generating, validating, and keeping track of large numbers of mutant animals is prohibitive. Here, we introduce Multiplexed Intermixed CRISPR Droplets (MIC-Drop), a platform combining droplet microfluidics, single-needle en masse CRISPR ribonucleoprotein injections, and DNA barcoding to enable large-scale functional genetic screens in zebrafish. The platform can efficiently identify genes responsible for morphological or behavioral phenotypes. In one application, we showed that MIC-Drop could identify small-molecule targets. Furthermore, in a MIC-Drop screen of 188 poorly characterized genes, we discovered several genes important for cardiac development and function. With the potential to scale to thousands of genes, MIC-Drop enables genome-scale reverse genetic screens in model organisms.
MeSH term(s)	Animals ; CRISPR-Cas Systems ; Cardiovascular System/growth & development ; Cell Culture Techniques ; Genetic Testing ; High-Throughput Nucleotide Sequencing ; Microfluidic Analytical Techniques ; Zebrafish/genetics ; Zebrafish/growth & development
Language	English
Publishing date	2021-08-19
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	128410-1
ISSN	1095-9203 ; 0036-8075
ISSN (online)	1095-9203
ISSN	0036-8075
DOI	10.1126/science.abi8870
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Zs.A 27: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
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Article ; Online: Multipotent Adult Progenitor Cells Support Lymphatic Regeneration at Multiple Anatomical Levels during Wound Healing and Lymphedema.

Beerens, Manu / Aranguren, Xabier L / Hendrickx, Benoit / Dheedene, Wouter / Dresselaers, Tom / Himmelreich, Uwe / Verfaillie, Catherine / Luttun, Aernout

Scientific reports

2018 Volume 8, Issue 1, Page(s) 3852

Abstract: Lymphatic capillary growth is an integral part of wound healing, yet, the combined effectiveness of stem/progenitor cells on lymphatic and blood vascular regeneration in wounds needs further exploration. Stem/progenitor cell transplantation also emerged ... ...

Abstract	Lymphatic capillary growth is an integral part of wound healing, yet, the combined effectiveness of stem/progenitor cells on lymphatic and blood vascular regeneration in wounds needs further exploration. Stem/progenitor cell transplantation also emerged as an approach to cure lymphedema, a condition caused by lymphatic system deficiency. While lymphedema treatment requires lymphatic system restoration from the capillary to the collector level, it remains undetermined whether stem/progenitor cells support a complex regenerative response across the entire anatomical spectrum of the system. Here, we demonstrate that, although multipotent adult progenitor cells (MAPCs) showed potential to differentiate down the lymphatic endothelial lineage, they mainly trophically supported lymphatic endothelial cell behaviour in vitro. In vivo, MAPC transplantation supported blood vessel and lymphatic capillary growth in wounds and restored lymph drainage across skin flaps by stimulating capillary and pre-collector vessel regeneration. Finally, human MAPCs mediated survival and functional reconnection of transplanted lymph nodes to the host lymphatic network by improving their (lymph)vascular supply and restoring collector vessels. Thus, MAPC transplantation represents a promising remedy for lymphatic system restoration at different anatomical levels and hence an appealing treatment for lymphedema. Furthermore, its combined efficacy on lymphatic and blood vascular growth is an important asset for wound healing.
MeSH term(s)	Animals ; Cell Culture Techniques ; Endothelial Cells/physiology ; Endothelium, Lymphatic ; Humans ; Lymph/physiology ; Lymph Nodes/physiopathology ; Lymphangiogenesis/physiology ; Lymphatic System/physiopathology ; Lymphatic Vessels/pathology ; Lymphedema/metabolism ; Lymphedema/pathology ; Lymphocytes/metabolism ; Mice ; Mice, Inbred C57BL ; Multipotent Stem Cells/metabolism ; Multipotent Stem Cells/physiology ; Stem Cell Transplantation/methods ; Stem Cells ; Wound Healing/physiology
Language	English
Publishing date	2018-03-01
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-018-21610-8
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