TY  - JOUR
AB  - Cardiosphere-derived cells (CDCs) generated from human cardiac biopsies have been shown to have disease-modifying bioactivity in clinical trials. Paradoxically, CDCs' cellular origin in the heart remains elusive. We studied the molecular identity of CDCs using single-cell RNA sequencing (sc-RNAseq) in comparison to cardiac non-myocyte and non-hematopoietic cells (cardiac fibroblasts/CFs, smooth muscle cells/SMCs and endothelial cells/ECs). We identified CDCs as a distinct and mitochondria-rich cell type that shared biological similarities with non-myocyte cells but not with cardiac progenitor cells derived from human-induced pluripotent stem cells. CXCL6 emerged as a new specific marker for CDCs. By analysis of sc-RNAseq data from human right atrial biopsies in comparison with CDCs we uncovered transcriptomic similarities between CDCs and CFs. By direct comparison of infant and adult CDC sc-RNAseq data, infant CDCs revealed GO-terms associated with cardiac development. To analyze the beneficial effects of CDCs (pro-angiogenic, anti-fibrotic, anti-apoptotic), we performed functional in vitro assays with CDC-derived extracellular vesicles (EVs). CDC EVs augmented in vitro angiogenesis and did not stimulate scarring. They also reduced the expression of pro-apoptotic Bax in NRCMs. In conclusion, CDCs were disclosed as mitochondria-rich cells with unique properties but also with similarities to right atrial CFs. CDCs displayed highly proliferative, secretory and immunomodulatory properties, characteristics that can also be found in activated or inflammatory cell types. By special culture conditions, CDCs earn some bioactivities, including angiogenic potential, which might modify disease in certain disorders.
AU  - Kogan, P.S.*
AU  - Wirth, F.*
AU  - Tomar, A.
AU  - Darr, J.
AU  - Teperino, R.
AU  - Lahm, H.*
AU  - Dreßen, M.*
AU  - Puluca, N.*
AU  - Zhang, Z.*
AU  - Neb, I.*
AU  - Beck, N.*
AU  - Luzius, T.*
AU  - de la Osa de la Rosa, L.*
AU  - Gärtner, K.
AU  - Hüls, C.
AU  - Zeidler, R.
AU  - Ramanujam, D.*
AU  - Engelhardt, S.*
AU  - Wenk, C.*
AU  - Holdt, L.M.*
AU  - Mononen, M.*
AU  - Sahara, M.*
AU  - Cleuziou, J.*
AU  - Hörer, J.*
AU  - Lange, R.*
AU  - Krane, M.*
AU  - Doppler, S.A.*
C1  - 64502
C2  - 51936
CY  - Tiergartenstrasse 17, D-69121 Heidelberg, Germany
TI  - Uncovering the molecular identity of cardiosphere-derived cells (CDCs) by single-cell RNA sequencing.
JO  - Basic Res. Cardiol.
VL  - 117
IS  - 1
PB  - Springer Heidelberg
PY  - 2022
SN  - 0300-8428
ER  - 

TY  - JOUR
AB  - Alterations of RNA editing that affect the secondary structure of RNAs can cause human diseases. We therefore studied RNA editing in failing human hearts. Transcriptome sequencing showed that adenosine-to-inosine (A-to-I) RNA editing was responsible for 80% of the editing events in the myocardium. Failing human hearts were characterized by reduced RNA editing. This was primarily attributable to Alu elements in introns of protein-coding genes. In the failing left ventricle, 166 circRNAs were upregulated and 7 circRNAs were downregulated compared to non-failing controls. Most of the upregulated circRNAs were associated with reduced RNA editing in the host gene. ADAR2, which binds to RNA regions that are edited from A-to-I, was decreased in failing human hearts. In vitro, reduction of ADAR2 increased circRNA levels suggesting a causal effect of reduced ADAR2 levels on increased circRNAs in the failing human heart. To gain mechanistic insight, one of the identified upregulated circRNAs with a high reduction of editing in heart failure, AKAP13, was further characterized. ADAR2 reduced the formation of double-stranded structures in AKAP13 pre-mRNA, thereby reducing the stability of Alu elements and the circularization of the resulting circRNA. Overexpression of circAKAP13 impaired the sarcomere regularity of human induced pluripotent stem cell-derived cardiomyocytes. These data show that ADAR2 mediates A-to-I RNA editing in the human heart. A-to-I RNA editing represses the formation of dsRNA structures of Alu elements favoring canonical linear mRNA splicing and inhibiting the formation of circRNAs. The findings are relevant to diseases with reduced RNA editing and increased circRNA levels and provide insights into the human-specific regulation of circRNA formation.
AU  - Kokot, K.E.*
AU  - Kneuer, J.M.*
AU  - John, D. *
AU  - Rebs, S.*
AU  - Möbius-Winkler, M.N.*
AU  - Erbe, S.*
AU  - Müller, M.*
AU  - Andritschke, M.*
AU  - Gaul, S.*
AU  - Sheikh, B.
AU  - Haas, J.*
AU  - Thiele, H.*
AU  - Müller, O.J.*
AU  - Hille, S.*
AU  - Leuschner, F.*
AU  - Dimmeler, S.*
AU  - Streckfuss-Bömeke, K.*
AU  - Meder, B.*
AU  - Laufs, U.*
AU  - Boeckel, J.N.*
C1  - 65536
C2  - 52318
TI  - Reduction of A-to-I RNA editing in the failing human heart regulates formation of circular RNAs.
JO  - Basic Res. Cardiol.
VL  - 117
IS  - 1
PY  - 2022
SN  - 0300-8428
ER  - 

TY  - JOUR
AB  - For a long time, gene editing had been a scientific concept, which was limited to a few applications. With recent developments, following the discovery of TALEN zinc-finger endonucleases and in particular the CRISPR/Cas system, gene editing has become a technique applicable in most laboratories. The current gain- and loss-of function models in basic science are revolutionary as they allow unbiased screens of unprecedented depth and complexity and rapid development of transgenic animals. Modifications of CRISPR/Cas have been developed to precisely interrogate epigenetic regulation or to visualize DNA complexes. Moreover, gene editing as a clinical treatment option is rapidly developing with first trials on the way. This article reviews the most recent progress in the field, covering expert opinions gathered during joint conferences on genome editing of the German Cardiac Society (DGK) and the German Center for Cardiovascular Research (DZHK). Particularly focusing on the translational aspect and the combination of cellular and animal applications, the authors aim to provide direction for the development of the field and the most frequent applications with their problems.
AU  - Brandes, R.P.*
AU  - Dueck, A.*
AU  - Engelhardt, S.*
AU  - Kaulich, M.*
AU  - Kupatt, C.*
AU  - De Angelis, M.T.*
AU  - Leisegang, M.S.*
AU  - le Noble, F.*
AU  - Moretti, A.*
AU  - Müller, O.J.*
AU  - Skryabin, B.V.*
AU  - Thum, T.*
AU  - Wurst, W.
C1  - 61057
C2  - 50021
CY  - Tiergartenstrasse 17, D-69121 Heidelberg, Germany
TI  - DGK and DZHK position paper on genome editing: Basic science applications and future perspective.
JO  - Basic Res. Cardiol.
VL  - 116
IS  - 1
PB  - Springer Heidelberg
PY  - 2021
SN  - 0300-8428
ER  - 

TY  - JOUR
AB  - Upon activation, neutrophils release neutrophil extracellular traps (NETs), which contribute to circulating DNA burden and thrombosis, including ST-segment elevation myocardial infarction (STEMI). Deoxyribonuclease (DNase) 1 degrades circulating DNA and NETs. Lower DNase activity correlates with NET burden and infarct size. The DNase 1 Q222R single nucleotide polymorphism (SNP), impairing DNase 1 function, is linked with myocardial infarction. We assessed whether the Q222R SNP is connected to increased NET burden in STEMI and influences long-term outcomes. We enrolled 711 STEMI patients undergoing primary percutaneous coronary intervention (pPCI), and 1422 controls. Genotyping was performed for DNase 1 Q222R SNP. DNase activity, double-stranded (ds)DNA and citrullinated histone H3 were determined in culprit site and peripheral plasma during pPCI. The association of the Q222R variant on cardiovascular and all-cause mortality was assessed by multivariable Cox regression adjusted for cardiovascular risk factors. Homozygous Q222R DNase 1 variant was present in 64 (9.0%) STEMI patients, at the same frequency as in controls. Patients homozygous for Q222R displayed less DNase activity and increased circulating DNA burden. In overall patients, median survival was 60 months. Homozygous Q222R variant was independently associated with cardiovascular and all-cause mortality after STEMI. dsDNA/DNase ratio independently predicted cardiovascular and all-cause mortality. These findings highlight that the Q222R DNase 1 SNP is associated with increased NET burden and decreased compensatory DNase activity, and may serve as an independent risk factor for poor outcome after STEMI.
AU  - Hofbauer, T.M.*
AU  - Mangold, A.*
AU  - Ondracek, A.S.*
AU  - Panzenböck, A.*
AU  - Scherz, T.*
AU  - Müller, J.*
AU  - Distelmaier, K.*
AU  - Seidl, V.*
AU  - Kastl, S.*
AU  - Müller-Nurasyid, M.
AU  - Peters, A.
AU  - Strauch, K.
AU  - Winker, R.*
AU  - Wohlschläger-Krenn, E.*
AU  - Nistler, S.*
AU  - Lang, I.M.*
C1  - 61885
C2  - 50501
CY  - Tiergartenstrasse 17, D-69121 Heidelberg, Germany
TI  - Deoxyribonuclease 1 Q222R single nucleotide polymorphism and long-term mortality after acute myocardial infarction.
JO  - Basic Res. Cardiol.
VL  - 116
IS  - 1
PB  - Springer Heidelberg
PY  - 2021
SN  - 0300-8428
ER  - 

TY  - JOUR
AB  - Cardiomyopathy is one of the most common causes of chronic heart failure worldwide. Mutations in the gene encoding nexilin (NEXN) occur in patients with both hypertrophic and dilated cardiomyopathy (DCM); however, little is known about the pathophysiological mechanisms and relevance of NEXN to these disorders. Here, we evaluated the functional role of NEXN using a constitutive Nexn knock-out (KO) mouse model. Heterozygous (Het) mice were inter-crossed to produce wild-type (WT), Het, and homozygous KO mice. At birth, 32, 46, and 22 % of the mice were WT, Het, and KO, respectively, which is close to the expected Mendelian ratio. After postnatal day 6, the survival of the Nexn KO mice decreased dramatically and all of the animals died by day 8. Phenotypic characterizations of the WT and KO mice were performed at postnatal days 1, 2, 4, and 6. At birth, the relative heart weights of the WT and KO mice were similar; however, at day 4, the relative heart weight of the KO group was 2.3-fold higher than of the WT group. In addition, the KO mice developed rapidly progressive cardiomyopathy with left ventricular dilation and wall thinning and decreased cardiac function. At day 6, the KO mice developed a fulminant DCM phenotype characterized by dilated ventricular chambers and systolic dysfunction. At this stage, collagen deposits and some elastin deposits were observed within the left ventricle cavity, which resembles the features of endomyocardial fibroelastosis (EFE). Overall, these results further emphasize the role of NEXN in DCM and suggest a novel role in EFE.
AU  - Aherrahrou, Z*
AU  - Schlossarek, S.*
AU  - Stoelting, S.*
AU  - Klinger, M.*
AU  - Geertz, B.*
AU  - Weinberger, F.*
AU  - Kessler, T.*
AU  - Aherrahrou, R.*
AU  - Moreth, K.
AU  - Bekeredjian, R.*
AU  - Hrabě de Angelis, M.
AU  - Just, S.*
AU  - Rottbauer, W.*
AU  - Eschenhagen, T.*
AU  - Schunkert, H.*
AU  - Carrier, L.*
AU  - Erdmann, J.*
C1  - 47539
C2  - 40658
CY  - Heidelberg
TI  - Knock-out of nexilin in mice leads to dilated cardiomyopathy and endomyocardial fibroelastosis.
JO  - Basic Res. Cardiol.
VL  - 111
IS  - 1
PB  - Springer Heidelberg
PY  - 2016
SN  - 0300-8428
ER  - 

TY  - JOUR
AB  - The platelet collagen receptor glycoprotein VI (GPVI) mediates platelet adhesion to subendothelial matrix and thrombus formation in acute coronary syndrome (ACS). This study examined patients with both ACS and stable coronary artery disease (CAD), which presented with atrial fibrillation (AF) and sinus rhythm (SR). We evaluated 992 patients with acute or stable CAD, and determined platelet surface expression of GPVI using flow cytometry. Seventy-eight patients presented with nonvalvular persistent AF. After 1:1 propensity score matching 156 matched cases with 78 pairs were obtained. Patients with AF and ACS showed a significantly decreased GPVI expression compared to patients with ACS and SR, whereas patients with stable angina pectoris (SA) presented with low level activation and no significant difference between SR and AF [mean fluorescence intensity (MFI) for ACS (SR Vs. AF): 20 +/- A 6.3 Vs. 17.7 +/- A 4.4; P = 0.023; SA (SR Vs. AF): 18.8 +/- A 9.4 Vs. 18.1 +/- A 6.1; P = 0.649]. In contrast, soluble GPVI was increased in ACS and AF accordingly [plasma GPVI (ng/ml) for ACS (SR Vs. AF): 1.4 +/- A 0.8 Vs. 1.9 +/- A 1.1; P = 0.038; SA (SR Vs. AF): 0.9 +/- A 0.4 Vs. 1.1 +/- A 0.5; P = 0.127]. Platelet GPVI surface expression is decreased in patients with AF and ACS compared to patients with SR and ACS. Nonvalvular AF is related to indices of chronic platelet activation and might be responsible for a down-regulation of GPVI receptor density on platelets, while soluble GPVI was increased in ACS and AF accordingly.
AU  - Bigalke, B.*
AU  - Stellos, K.*
AU  - Weig, H.J.*
AU  - Geisler, T.*
AU  - Seizer, P.*
AU  - Kremmer, E.
AU  - Pötz, O.*
AU  - Joos, T.*
AU  - May, A.E.*
AU  - Lindemann, S.*
AU  - Gawaz, M.*
C1  - 1667
C2  - 26509
SP  - 352-357
TI  - Regulation of platelet glycoprotein VI (GPVI) surface expression and of soluble GPVI in patients with atrial fibrillation (AF) and acute coronary syndrome (ACS).
JO  - Basic Res. Cardiol.
VL  - 104
IS  - 3
PB  - Dietrich Steinkopff
PY  - 2009
SN  - 0300-8428
ER  - 

TY  - JOUR
AB  - Platelet adhesion to the atherosclerotic vascular wall induces thrombosis and boosters vascular inflammation and atheroprogression. In the present study we studied the binding of the platelet collagen receptor glycoprotein (GP) VI to human atherosclerotic plaques (AP) and the role of GPVI-mediated platelet adhesion for atheroprogression. Soluble GPVI-Fc fusion protein bound to immobilized collagen type I, collagen type III, and predominantly to the core region of human carotid atheromatous plaques. The pattern of GPVI-Fc binding was similar to the immunostaining pattern of collagen type III and differed from the immunostaining of collagen type I, which was more intense in the cap than in the core. Plaque-induced platelet aggregation in stirred blood and platelet adhesion/aggregate formation under flow were inhibited by the anti-GPVI monoclonal antibody 5C4 or by pretreatment of plaques with anti-collagen type I and anti-collagen type III antibody, or GPVI-Fc. However, there was no correlation between GPVI-Fc binding and platelet aggregating activity of individual plaques. GPVI bound also to atherosclerotic arteries of ApoE-deficient mice in vivo as assessed by small animal positron emission tomography (PET). Prolonged administration of soluble GPVI attenuated atheroprogression in ApoE-deficient mice. In humans, GPVI binding to collagenous type I and type III structures of the plaque core region mediates plaque-induced platelet adhesion and aggregation, but GPVI binding is not the sole platelet-activating determinant of plaques. In mice, GPVI-mediated platelet adhesion to the atherosclerotic vascular wall is involved in atheroprogression in vivo. Taken together, our data suggests that GPVI is a relevant target to prevent atherothrombotic events and atheroprogression.
AU  - Schulz, C.*
AU  - Penz, S.*
AU  - Hoffmann, C.*
AU  - Langer, H.*
AU  - Gillitzer, A.*
AU  - Schneider, S.*
AU  - Brandl, R.*
AU  - Seidl, S.*
AU  - Massberg, S.*
AU  - Pichler, B.*
AU  - Kremmer, E.
AU  - Stellos, K.*
AU  - Schönberger, T.*
AU  - Siess, W.*
AU  - Gawaz, M.*
C1  - 693
C2  - 25849
SP  - 356-367
TI  - Platelet GPVI binds to collagenous structures in the core region of human atheromatous plaque and is critical for atheroprogression in vivo.
JO  - Basic Res. Cardiol.
VL  - 103
IS  - 4
PB  - Steinkopff
PY  - 2008
SN  - 0300-8428
ER  -