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Cardiovascular diseases remain a major health burden in nowadays society. Patients have to undergo surgery due to the lack of pharmaceutical treatment. Here, we show how therapeutic modulation of the long non-coding RNA NUDT6 provides a promising strategy to treat both, carotid artery disease and abdominal aortic aneurysms. NUDT6 was identified in a transcriptomic array of stable versus ruptured fibrous caps from human carotid artery plaques. NUDT6 regulates vascular smooth muscle cell survival and migration which are crucial processes in the above-mentioned diseases., Herz-Kreislauf-Erkrankungen stellen in der heutigen Gesellschaft nach wie vor eine große Belastung dar. Patienten müssen sich operiert werden, da es keine andere Behandlung gibt. Hier zeigen wir, wie die therapeutische Modulation der langen nichtkodierenden RNA NUDT6 eine neue Art der Behandlung von Erkrankungen der Halsschlagader und abdominalen Aortenaneurysmen darstellt. NUDT6 wurde in einem transkriptomischen Array von stabilen und rupturierten fibrösen Kappen aus menschlichen Karotis-Plaques identifiziert. NUDT6 reguliert das Überleben und die Migration von glatten Gefäßmuskelzellen, die bei den genannten Krankheiten eine wichtige Rolle spielen.

The aorta-on-a-chip (AoOC) is a micro-engineered 3D in vitro model which aims to mimic the in vivo complexity of a human aorta in terms of cell-cell interaction, tissue architecture and hemodynamic conditions, i.e., the wall shear stress (WSS) force that the blood exerts on the vessel luminal surface in the direction of blood flow. Transcriptomic changes in primary aortic endothelial cells and primary aortic smooth muscle cells due to different flow conditions were identified using RNA sequencing analysis., Die Aorta-on-a-chip (AoOC) ist ein mikrotechnisch entwickeltes 3D-In-vitro-Modell, welches die in vivo-Komplexität einer menschlichen Aorta in Bezug auf Zell-Zell-Interaktion, Gewebearchitektur und hämodynamische Bedingungen, d. h. die vom Blut auf die Gefäßlumina in Richtung des Blutflusses ausgeübte Wandschubspannung (WSS), nachbilden soll. Anhand von RNA-Sequenzierungsanalysen wurden transkriptomische Veränderungen in primären Aortenendothelzellen und primären glatten Muskelzellen der Aorta infolge unterschiedlicher Strömungsbedingungen identifiziert.

Through proteomic sequencing of surgially obtained atherosclerotic plaque lesions AIFM1 was indentified as a potential biomarker indicating plaque stability in its downregulated presence and instability in its upregulated presence. Investigating via various scientific experiments a clear trend regarding this hypothesis was determined. The advantage of biobanking and proteomic sequencing dealing with potential biomarker for cardiovascular diseases was indicated., Durch Sequenzierung von chirurgisch aquirierten Läsionen der Karotiden wurde AIFM1 als potentieller Biomarker hinsichtlich Differenzierung stabiler Plaques bei dessen Hochregulierung versus instabiler atherosklerotischen Läsionen bei dessen Herunterregulierung indentifiziert. Durch weitere Experiemente konnte eine klarer Trend zu dieser These gefunden werden. Der Vorteil von Biobanken und der Proteom Sequenzierung zur Detektion potenzieller Biomarker für Kardiovaskuläre Erkrankungen wurde klar.

Vascular smooth muscle cells maintain plasticity and contribute to vulnerable plaque formation in carotid artery disease. The study aimed to establish a vascular cell biobank to examine the phenotype and protein expression of VSMCs as the disease progresses from stable to ruptured lesions. Carotid artery stenosis samples were collected during carotid endarterectomy and the VSMCs were isolated and cultured. The cell-state was indirectly analysed by establishing a gene panel. Direct analysis was performed with live-cell imaging for migration and proliferation behaviour., Die Plastizität der glatten Gefäßmuskelzellen ist essenziell in der Entstehung von vulnerablen Plaques in Karotisstenosen. Diese Studie beabsichtigt die Etablierung einer vaskulären Zellbiobank zur Phänotypanalyse der glatten Gefäßmuskelzellen in stabilen vs rupturierten Plaques nach Karotisendarteriektomie. Mit der Etablierung einer Genexpressionsanalyse wurde der Phänotyp indirekt anhand kontraktiler und synthetischer Gene bestimmt. Für eine direkte Analyse wurde eine Live-cell Imaging Analyse der Migrations- und Proliferationsaktivität durchgeführt.

Background: Leukocyte progenitors derived from clonal hematopoiesis of undetermined potential (CHIP) are associated with increased cardiovascular events. However, the prevalence and functional relevance of CHIP in coronary artery disease (CAD) are unclear, and cells affected by CHIP have not been detected in human atherosclerotic plaques. Methods: CHIP mutations in blood and tissues were identified by targeted deep-DNA-sequencing (DNAseq: coverage >3,000) and whole-genome-sequencing (WGS: coverage >35). CHIP-mutated leukocytes were visualized in human atherosclerotic plaques by mutaFISHTM. Functional relevance of CHIP mutations was studied by RNAseq. Results: DNAseq of whole blood from 540 deceased CAD patients of the Munich cardIovaScular StudIes biObaNk (MISSION) identified 253 (46.9%) CHIP mutation carriers (mean age 78.3 years). DNAseq on myocardium, atherosclerotic coronary and carotid arteries detected identical CHIP mutations in 18 out of 25 mutation carriers in tissue DNA. MutaFISHTM visualized individual macrophages carrying DNMT3A CHIP mutations in human atherosclerotic plaques. Studying monocyte-derived macrophages from Stockholm-Tartu Atherosclerosis Reverse Networks Engineering Task (STARNET; n=941) by WGS revealed CHIP mutations in 14.2% (mean age 67.1 years). RNAseq of these macrophages revealed that expression patterns in CHIP mutation carriers differed substantially from those of non-carriers. Moreover, patterns were different depending on the underlying mutations, e.g. those carrying TET2 mutations predominantly displayed upregulated inflammatory signaling whereas ASXL1 mutations showed stronger effects on metabolic pathways. Conclusions: Deep-DNA-sequencing reveals a high prevalence of CHIP mutations in whole blood of CAD patients. CHIP-affected leukocytes invade plaques in human coronary arteries. RNAseq data obtained from macrophages of CHIP-affected patients suggest that pro-atherosclerotic signaling differs depending on the underlying mutation

Das GUCY1A1 Gen wurde in genomweiten Assoziationsstudien mit der koronaren Herzkrankheit (KHK) assoziiert. Die Risikovariante rs7692387 führte zu reduzierter Expression von GUCY1A1 und nach Hinzufügen von NO zu reduzierter Inhibierung der Thrombozytenaggregation bei homozygoten Risikoallelträgern. Dies wurde durch Präinkubation mit Aspirin aufgehoben. Aspirin kann ein Therapieansatz bei homozygoten GUCY1A1 Risikoallelträgern zur Reduktion des KHK-Risikos sein., The GUCY1A1 gene has been associated with coronary artery disease (CAD) by genome-wide association studies. The risk variant rs7692387 leads to reduced expression of GUCY1A1 and after the addition of NO to reduced inhibition of platelet aggregation in homozygous risk allele carriers, which was reversed by pre-incubation with aspirin. Aspirin might be a therapeutic strategy in homozygous GUCY1A1 risk allele carriers to reduce CAD risk.

Leitlinien empfehlen die operative Therapie (CEA) höhergradiger Carotisstenosen vorausgesetzt das perioperative Schlaganfall-/Todesrisiko ist niedrig. Die externe Qualitätssicherung in Deutschland erfasst diese Komplikationen während des Krankenhausaufenthalts. Diese Studie untersuchte Schlaganfälle/Todesfälle im 30-Tage-Intervall nach CEA. Ca. 60% der Schlaganfälle/Todesfälle traten stationär und ca. 40% poststationär auf. Die frühe poststationäre Nachsorge sollte daher intensiviert werden., Guidelines recommend operative treatment (CEA) of high-grade carotid artery stenosis providing that the perioperative risk of stroke or death is low. Within the german mandatory quality assurance these complications are recorded for the in-hospital stay. This study examined strokes/deaths within 30 days after CEA. About 60% of all post-operative strokes/deaths occured during in-hospital stay and about 40% after discharge. Early post-hospital follow-up care should therefore be intensified.

Targeting inflammation in atherosclerosis with colchicine represents a novel therapeutical approach. The aim of this dissertation is to elucidate the biological basis of this beneficial effect. In this context, it was shown that colchicine treatment decreases vascular inflammation, and thus the progression of atherosclerosis, by reducing the uptake of inflammatory leukocytes into atherosclerotic plaques., Die anti-inflammatorische Therapie der Atherosklerose mit Colchicin repräsentiert eine neue Behandlungsstrategie. Das Ziel dieser Dissertation ist es, die biologischen Grundlagen dieses präventiven Effekts näher zu entschlüsseln. In diesem Zusammenhang konnte gezeigt werden, dass die Behandlung mit Colchicin die vaskuläre Entzündung und damit das Fortschreiten der Atherosklerose abschwächt, insbesondere durch verminderte Aufnahme von entzündlichen Leukozyten in atherosklerotische Plaques.

An Patienten, die sich einem endovaskulären Aortenrepair unterzogen, wurden Schwankungen der Konzentrationen der Neuropeptide Oxytocin (OXT) und Arginin-Vasopressin (AVP) in Blut, Speichel und Liquor untersucht. Ergebnisse: OXT-Konzentrationen im Liquor und Speichel fielen intraoperativ. OXT-Blutkonzentrationen stiegen präoperativ. Zu allen Zeitpunkten zeigte sich eine Korrelation von OXT Liquor- und Speichelkonzentrationen. Diese war stärker als zwischen Liquor und Blut. AVP im Blut zeigte eine inverse Beziehung zu den Angst- und Depressions-Scores., Fluctuations in the concentrations of the neuropeptides oxytocin (OXT) and arginine vasopressin (AVP) in blood, saliva, and cerebrospinal fluid (CSF) were studied in patients undergoing endovascular aortic repair. Results: OXT concentrations in CSF and saliva fell intraoperatively. OXT blood concentrations increased preoperatively. At all time points, there was a correlation of OXT CSF and saliva concentrations. This was stronger than between CSF and blood. AVP in blood showed an inverse relationship with anxiety and depression scores.

Long non-coding RNAs (lncRNAs) orchestrate various biological processes and regulate the development of cardiovascular diseases. Their potential therapeutic benefit to tackle disease progression has recently been extensively explored. Our study investigates the role of lncRNA Nudix Hydrolase 6 (NUDT6) and its antisense target fibroblast growth factor 2 (FGF2) in two vascular pathologies: abdominal aortic aneurysms (AAA) and carotid artery disease. Using tissue samples from both diseases, we detected a substantial increase of NUDT6, whereas FGF2 was downregulated. Targeting Nudt6 in vivo with antisense oligonucleotides in three murine and one porcine animal model of carotid artery disease and AAA limited disease progression. Restoration of FGF2 upon Nudt6 knockdown improved vessel wall morphology and fibrous cap stability. Overexpression of NUDT6 in vitro impaired smooth muscle cell (SMC) migration, while limiting their proliferation and augmenting apoptosis. By employing RNA pulldown followed by mass spectrometry as well as RNA immunoprecipitation, we identified Cysteine and Glycine Rich Protein 1 (CSRP1) as another direct NUDT6 interaction partner, regulating cell motility and SMC differentiation. Overall, the present study identifies NUDT6 as a well-conserved antisense transcript of FGF2. NUDT6 silencing triggers SMC survival and migration and could serve as a novel RNA-based therapeutic strategy in vascular diseases.

An abdominal aortic aneurysm (AAA) is a pathological widening of the aortic wall characterized by loss of smooth muscle cells (SMCs), extracellular matrix degradation, and local inflammation. This condition is often asymptomatic until rupture occurs, leading to high morbidity and mortality rates. Diagnosis is mostly accidental and the only currently available treatment option remains surgical intervention. Circular RNAs (circRNAs) represent a novel class of regulatory non-coding RNAs that originate from backsplicing. Their highly stable loop structure, combined with a remarkable enrichment in body fluids, make circRNAs promising disease biomarkers. We investigated the contribution of circRNAs to AAA pathogenesis and their potential application to improve AAA diagnostics. Gene expression analysis revealed the presence of deregulated circular transcripts stemming from AAA-relevant gene loci. Among these, the circRNA to the Ataxia Telangiectasia Mutated gene (cATM) was upregulated in human AAA specimens, in AAA-derived SMCs, and serum samples collected from aneurysm patients. In primary aortic SMCs, cATM increased upon angiotensin II and doxorubicin stimulation, while its silencing triggered apoptosis. Higher cATM levels made AAA-derived SMCs less vulnerable to oxidative stress, compared with control SMCs. These data suggest that cATM contributes to elicit an adaptive oxidative-stress response in SMCs and provides a reliable AAA disease signature.

Highlights • Endothelial ageing contributes significantly to atherosclerosis. • Non-coding RNAs are gaining interest as regulators of vascular biology. • Several microRNAs regulate endothelial cell ageing. • Multiple lncRNAs play a role in endothelial cell ageing. Abstract Atherosclerosis and numerous other cardiovascular diseases develop in an age-dependent manner. The endothelial cells that line the vessel walls play an important role in the development of atherosclerosis. Non-coding RNA like microRNAs and long non-coding RNAs are known to play an important role in endothelial function and are implicated in the disease progression. Here, we summarize several microRNAs and long non-coding RNAs that are known to have an altered expression with endothelial aging and discuss their role in endothelial cell function and senescence. These processes contribute to aging-induced atherosclerosis development and by targeting the non-coding RNAs controlling endothelial cell function and senescence, atherosclerosis can potentially be attenuated.

Abdominal aortic aneurysm (AAA) poses a great threat to patients. The aim of my thesis project was to reveal the cellular heterogeneity of AAA at the single-cell level. Human AAA samples were collected from patients undergoing open repair surgery and were performed with 10X Genomics Chromium Platform. Current single-cell data analysis of these samples revealed the heterogeneity in AAA and characterized the subpopulations of macrophages and vascular smooth muscle cells, along with their possible functions during the AAA development and progression., Das abdominale Aortenaneurysma (AAA) stellt eine große Bedrohung für die Patienten dar. Das Ziel meines Dissertationsprojektes war es, die zelluläre Heterogenität des AAA auf Einzelzellebene aufzudecken.Menschliche AAA-Proben wurden von Patienten entnommen, die sich einer offenen Aortenoperation unterzogen, und mit der 10X Genomics Chromplattform durchgeführt. Die aktuelle Analyse der Einzelzelldaten dieser Proben ergab die Heterogenität des AAA und charakterisierte die Subpopulationen von Makrophagen und vaskuläre glatte Muskelzellen sowie ihre möglichen Funktionen während der Entwicklung und Progression des AAA.

Abdominal aortic aneurysm (AAA) is a life-threatening vascular disease, involving complex molecular and pathophysiological mechanisms. Smooth muscle cell (SMC) depletion appears as one of the specific characteristics in AAA progression. Necroptosis and apoptosis of SMC are contributing to aneurysm dilatation, and neutrophils extracellular traps (NETs) seem to propagate these effects. Targeting NETs could thus be a promising future therapeutic strategy to limit AAA disease., Das abdominale Aortenaneurysma (AAA) ist eine lebensbedrohliche Gefäßerkrankung mit komplexen molekularen und pathophysiologischen Mechanismen. Der Abbau der glatten Muskelzellen (SMC) erscheint als eines der spezifischen Merkmale bei der AAA-Progression. Nekroptose und Apoptose von SMC tragen zur Aneurysmadilatation bei, und die extrazellulären Neutrophilenfallen (NETs) scheinen diese Effekte zu verbreiten. Das Targeting von NETs könnte daher eine vielversprechende zukünftige Therapiestrategie zur Begrenzung der AAA-Krankheit sein.

Da die Prozesse der Mechanotransduktion weiterhin Gegenstand aktueller Forschung sind, ist es im Tissue Engineering essentiell, möglichst physiologische Bedingungen zu schaffen. Dafür wurde ein Verfahren zur automatisierten Dezellularisierung eines murinen Scaffolds entwickelt, das anschließend in einer dynamischen Kultur mit humanen Osteoblasten rezellularisiert wird. Um die Auswirkungen einer mechanischen Stimulation auf Osteoblasten nach Rezellularisierung zu erforschen, wurde eine mechanische Stimulationseinheit erfolgreich implementiert und angepasst., Since the processes of mechanotransduction continue to be the focus of current research, it is essential in the field of tissue engineering to establish a physiological environment. For this purpose, a method for automated decellularization of a murine scaffold was developed, which was subsequently recellularized in a dynamic culture with human osteoblasts. In order to investigate the effects of mechanical stimulation on osteoblasts after recellularization, a mechanical stimulation unit was successfully implemented and adapted.

Die für diese Arbeit durchgeführte Studie vergleicht verschiedene Methoden den Ballon in einer Herzkatheteruntersuchung an die Inflationsspritze anzuschließen und zu entlüften. Die Entlüftung über einen Dreiwegehahn führt zu signifikant weniger häufig Luft im Ballon als der direkte Anschluss. Schließt man den Katheter direkt an die Inflationsspritze an, macht es keinen Unterschied, ob das Ballonlumen im Vorfeld mit Inflationslösung gespült wird oder nicht., This study compares different methods of connecting PCI balloons and the inflation syringe while removing air from the balloon. The connection with a three-way valve achieved significantly less air in the inflated balloon as compared with direct connection. The direct connection showed no significant difference between purging and filling the lumen prior to making the connection or not.

Ein akuter Myokardinfarkt führt im Zuge der systemischen Immunantwort zur Bereitstellung von phänotypisch verschiedenen Blut-Monozyten. Klassische, inflammatorische (CD14-positive) Blut-Monozyten von Patienten mit akutem STEMI zeigen signifikante Genexpressionsänderungen mit zunächst pro-inflammatorischen und später pro-reparativen Veränderungen. Insbesondere die differentielle Expression der Gene CYP19A1, THBS1, CLU, TRPM6, MTUS1, CRIP2, IL1R2, IL1R1, IL1RAP, IL18R1, IL18RAP, IL21R, CX3CR1 deutet auf ihre Regulation der monozytären Immunantwort hin., Acute myocardial infarction induces the supply of phenotypically different blood-monocytes during the following systemic inflammatory response. Classical, inflammatory (CD14-positive) blood monocytes from patients with acute STEMI show significant changes in gene expression and first pro-inflammatory and later pro-reparative adaptions. Specifically, differential expression of genes CYP19A1, THBS1, CLU, TRPM6, MTUS1, CRIP2, IL1R2, IL1R1, IL1RAP, IL18R1, IL18RAP, IL21R and CX3CR1 indicates their importance for the regulation of the monocytic immune response.

Da die Prozesse der Mechanotransduktion weiterhin Gegenstand aktueller Forschung sind, ist es im Tissue Engineering essentiell, möglichst physiologische Bedingungen zu schaffen. Dafür wurde ein Verfahren zur automatisierten Dezellularisierung eines murinen Scaffolds entwickelt, das anschließend in einer dynamischen Kultur mit humanen Osteoblasten rezellularisiert wird. Um die Auswirkungen einer mechanischen Stimulation auf Osteoblasten nach Rezellularisierung zu erforschen, wurde eine mechanische Stimulationseinheit erfolgreich implementiert und angepasst., Since the processes of mechanotransduction continue to be the focus of current research, it is essential in the field of tissue engineering to establish a physiological environment. For this purpose, a method for automated decellularization of a murine scaffold was developed, which was subsequently recellularized in a dynamic culture with human osteoblasts. In order to investigate the effects of mechanical stimulation on osteoblasts after recellularization, a mechanical stimulation unit was successfully implemented and adapted.

Congenital heart diseases are a difficult target for clinical and developmental studies. Particularly, the investigation of pathophysiologic changes in early stages of cardiogenesis is vastly limited because of the restricted access to patient material at different stages of human development. The advent of hiPSC technology and the possibility of generating patient- specific hiPSC-cardiomyocytes solved the obstacles of limited tissue access to some extent, providing a new platform for molecular investigation. Hypoplastic left heart syndrome (HLHS) is a congenital cardiac malformation in which the heart consists of a single pumping chamber responsible for the body circulation, consisting of the right ventricle, whereas passive venous flow to the lungs maintains the pulmonary circulation. In several hiPSC-CM models recapitulating common HLHS mutations, important findings suggest an underlying cell cycle defect and an immature phenotype of diseased HLHS cardiomyocytes (CM). Nevertheless, a major drawback of hiPSC-derived cardiomyocytes (hiPSC-CM) is their lack of the mature subcellular organization, electromechanical properties, and three-dimensionality that are seen in differentiated human cardiomyocytes. These immature properties of standard-cultured hiPSC-CMs consequently limit the exploration of disease phenotypes. In the current study, hiPSC-CMs were successfully seeded onto decellularized non-human primate ventricular myocardium (cECM), thereby generating 3D heart patches. By subjecting these 3D heart patches to specialized biomimetic culture conditions consisting of continuous mechanical preload and electrical field stimulation, a significant increase in structural and functional maturation of hiPSC-CM was achieved. By partially overcoming the limitation of cellular immaturity, 3D cECM heart patches were generated from healthy and diseased hiPSC-CMs of three different HLHS patient lines. The dynamic readout of contraction force and beating rate analysis revealed, Seit Beginn der erfolgreichen Differenzierung von humanen induzierten pluripotenten Stammzellen (hiPSC) zu Kardiomyozyten (hiPSC-CM) wurde eine neuartige molekulare Untersuchung verschiedenster genetischer Mutationen im Kontex von intakten, lebenden humanen Zellen möglich, welches das Krankheitsverständnis vertieft und die Identifizierung pathophysiologischer Vorgänge unterschiedlichster kardiovaskulärer Erkrankungen ermöglicht hat. Das hiPSC-CM Zellmodell bietet sich auch für die Erforschung von kongenitalen Herzerkrankungen an. Diese sind meist heterogene Krankheitsentitäten, die kardiovaskuläre Malformationen während der embryonalen Entwicklung umfassen und die unterschiedlichsten Mutationen und klinischen Ausprägungen beinhalten. Zu dieser Gruppe zählt auch das hypoplastische Linksherzsyndrom (HLHS). Hierbei handelt es sich um eine kongenitale kardiale Malformation, bei der das Herz aus einer einzigen Kammer besteht, die für die Kreislaufversorgung verantwortlich ist. Diese Kammer wird dabei klassischerweise vom rechten Ventrikel gebildet, da der linke Ventrikel nur hypoplastisch und weitestgehend funktionslos angelegt ist. Der zugrunde liegende Pathomechanismus sowie eindeutige krankheitsverursachende Mutationen sind noch nicht im Detail erforscht, weshalb verschiedenste Hypothesen zum Pathomechanismus des HLHS bestehen. Durch die molekulare Aufarbeitung verschiedener HLHS-Patienten im hiPSC- Modell konnte bereits ein besseres Verständnis der Zelldefekte erzielt werden. Nichtsdestotrotz ist die Erforschung des Krankheitsphänotypes dadurch limitiert, dass hiPSC-CM einen imaturen Phänotyp besitzen, der nicht alle Funktionen von reifen Kardiomyozyten rekapitulieren kann. In der für diese Arbeit durchgeführte Studie wurden hiPSC von gesunden Probanden sowie drei verschiedene HLHS-Patientenlinien erfolgreich zu Kardiomyozyten differenziert. Auf der Basis von dezellularisiertem Primaten-Myokard wurde anschließend ein 3D-Herzpatch generiert, das eine verbesserte Matur

Patients with type 2 diabetes (T2D) are threatened by excessive cardiovascular morbidity and mortality. While accelerated arterial stiffening may represent a critical mechanistic factor driving cardiovascular risk in T2D, specific therapies to contain the underlying diabetic arterial remodeling have been elusive. The present translational study investigates the role of microRNA-29b (miR-29b) as a driver and therapeutic target of diabetic aortic remodeling and stiffening. Using a murine model (db/db mice), as well as human aortic tissue samples, we find that diabetic aortic remodeling and stiffening is associated with medial fibrosis, as well as fragmentation of aortic elastic layers. miR-29b is significantly downregulated in T2D and miR-29b repression is sufficient to induce both aortic medial fibrosis and elastin breakdown through upregulation of its direct target genes COL1A1 and MMP2 thereby increasing aortic stiffness. Moreover, antioxidant treatment restores aortic miR-29b levels and counteracts diabetic aortic remodeling. Concluding, we identify miR-29b as a comprehensive—and therefore powerful—regulator of aortic remodeling and stiffening in T2D that moreover qualifies as a (redox-sensitive) target for therapeutic intervention.

Chronische Hypoxie induziert pulmonale Hypertonie, pulmonalvaskuläres Remodeling und rechtsventrikuläre Hypertrophie. Hypoxie-induzierbare Faktoren (HIF) sind zentrale Regulatoren der Transkription in hypoxischen Zellen. Obwohl glatte Muskelzellen wesentlich zur Pathogenese der pulmonalen Hypertension beitragen, ist die Rolle von glattmuskulärem HIF unvollständig geklärt. Die vorliegende Arbeit zeigt, dass HIF1α und HIF2α in glatten Muskelzellen die Entwicklung von pulmonaler Hypertonie und pulmonalvaskulärem Remodeling nach chronischer Hypoxie fördern., Chronic hypoxia induces pulmonary hypertension, pulmonary vascular remodeling and right ventricular hypertrophy. Hypoxia-inducible factors (HIF) are master regulators of transcription in hypoxic cells. Although smooth muscle cells critically contribute to the pathogenesis of pulmonary hypertension, the specific role of smooth muscle HIF is not well understood. This study shows that the presence of HIF1α and HIF2α in smooth muscle cells promotes the development of hypoxia-induced pulmonary vascular remodeling and pulmonary hypertension.

Das A disintegrin and metalloproteinase with thrombospondin motifs 7- (ADAMTS7) Gen ist mit der Koronaren Herzkrankheit assoziiert. Diese Arbeit umfasst eine funktionelle und phänotypische Charakterisierung ADAMTS-7-defizienter humaner glatter Gefäßmuskelzellen, Untersuchungen zur ADAMTS7-Expressionsregulation und eine Evaluation spezifischer Antikörper. ADAMTS-7-Defizienz verminderte die Zellmigration unter proatherogenen Bedingungen nicht. Oxidiertes Low-density lipoprotein bewirkte eine Reduktion der ADAMTS7-Expression, welche durch Knockdown des Oxidized low-density lipoprotein receptor 1 teilweise reversibel war., The A disintegrin and metalloproteinase with thrombospondin motifs 7 (ADAMTS7) gene is associated with coronary artery disease. This work comprises a functional and phenotypical characterization of ADAMTS-7 deficient human vascular smooth muscle cells, analyses of ADAMTS7 gene expression regulation as well as an evaluation of specific antibodies. ADAMTS-7 deficiency did not impair cell migration under proatherogenic conditions. ADAMTS7 expression was reduced by oxidized Low-density lipoprotein, which was partly reversible by Oxidized low-density lipoprotein receptor 1 knockdown.

Abdominal aortic aneurysm (AAA) is a life-threatening vascular disease, involving complex molecular and pathophysiological mechanisms. Smooth muscle cell (SMC) depletion appears as one of the specific characteristics in AAA progression. Necroptosis and apoptosis of SMC are contributing to aneurysm dilatation, and neutrophils extracellular traps (NETs) seem to propagate these effects. Targeting NETs could thus be a promising future therapeutic strategy to limit AAA disease., Das abdominale Aortenaneurysma (AAA) ist eine lebensbedrohliche Gefäßerkrankung mit komplexen molekularen und pathophysiologischen Mechanismen. Der Abbau der glatten Muskelzellen (SMC) erscheint als eines der spezifischen Merkmale bei der AAA-Progression. Nekroptose und Apoptose von SMC tragen zur Aneurysmadilatation bei, und die extrazellulären Neutrophilenfallen (NETs) scheinen diese Effekte zu verbreiten. Das Targeting von NETs könnte daher eine vielversprechende zukünftige Therapiestrategie zur Begrenzung der AAA-Krankheit sein.

Hintergrund: Die Entstehung von In-Stent Restenose (ISR) stellt eine der wichtigsten Limitationen aktueller Koronarstent-Technologie dar. Intravaskuläre Bildgebungstechnologien, in diesem Fall insbesondere die optische Kohärenztomografie (Optical Coherence Tomography, OCT), liefern wertvolle zusätzliche Informationen für die ISR-Charakterisierung, die bei einer Angiografie nicht verfügbar sind. Zweck: Zweck dieser Studie war die Durchführung einer qualitativen und quantitativen neointimalen Charakterisierung von ISR-Läsionen. Es erfolgt die Ermittlung der pathophysiologischen Ursache der In-Stent Restenose durch neue OCT-Surrogat Parameter innerhalb gestenteter Gefäßabschnitte, mittels neuen multimodalem Beurteilungsprotokoll. Ein weiterer wichtiger Punkt ist die Identifikation von Prädiktoren von neointimalen Mustern und Neoatherosklerose. Methodik: In dieser Studie wurden Patienten, die sich einer frequenzbasierten OCT (Frequency domain OCT, FD-OCT) vor einer perkutanen Koronarintervention (PCI) bei ISR unterzogen, an drei europäischen Prüfzentren eingeschlossen. Qualitative und morphometrische OCT-Analysen wurden in einem Core Lab jeder Millimeter entlang des gesamten gestenteten Segments durchgeführt. Weitere qualitative und quantitative Analysen in Bezug auf die Grauskala-Signalintensität (GSI) wurden pro Quadrant beim Einzelbild mit dem maximalen Prozentbereich der Stenose (=%AS-Wert) sowie bei den fünf vorangegangenen oder darauffolgenden analysierten Einzelbildern durchgeführt. Ergebnisse: Insgesamt nahmen an der Studie 107 Patienten teil, wobei pro Patient eine Läsion bildgebungstechnisch erfasst und behandelt wurde. Vorwiegend homogene Läsionen wiesen 4,5 % (0,0–14,3) nicht-homogene Quadranten auf, wohingegen vorwiegend nicht-homogene Läsionen 28,1 % (20,3–37,5) homogene Quadranten aufwiesen. Die durchschnittlichen GSI-Werte unterschieden sich erheblich zwischen homogenen [108,4 (92,5–123,6) U], nicht-homogenen [79,9 (61,2–95,9), Background: In-stent Restenosis (ISR) represents a major cause of stent failure. Intravascular imaging techniques under which here mostly to mention Optical coherence tomography (OCT) provides valuable unique additional Information for the characterization of ISR, otherwise not available on angiography. Purpose: To perform a qualitative and quantitative neointimal characterization of ISR lesions and identify predictors of neointimal patterns and neoatherosclerosis. Methods: Patients undergoing frequency-domain OCT prior to Percutaneous Coronary Intervention (PCI) for ISR in 3 European centres were included. Qualitative and morphometric OCT analyses were performed in a core laboratory every Millimeter along the entire stented segment. Additional qualitative and quantitative [gray-scale signal intensity (GSI)] analyses were performed on a per quadrant basis at the frame displaying the maximal percentage area stenosis (%AS) as well as the five preceding and following analyzed frames. Results: A total of 107 patients were included with one lesion being imaged and treated per patient. Predominantly homogeneous lesions included 4.5 (0.0-14.3) % non-homogeneous quadrants while predominantly non-homogeneous ones included 28.1 (20.3-37.5) % homogeneous quadrants. Mean GSI values were significantly different between homogeneous [108.4 (92.5-123.6) U], non-homogeneous [79.9 (61.2-95.9) U] and neoatherosclerosis [88.3 (72.8-104.9) U] quadrants (p<0.001 for all between group comparisons). Stent underexpansion (defined as stent expansion index <0.80) was observed in 16 (48.5%) and 44 (61.1%) of predominantly non-homogenenous and homogeneous ISR lesions respectively (p=0.225). Presence of any frame with neoatherosclerosis was registered in 33 (30.8%) ISR lesions. Multivariable analysis showed that female sex, maximal neointimal thickness, angiographic ISR pattern and diabetes mellitus independently correlate with neointimal pattern. Among all analyzed v

Das A disintegrin and metalloproteinase with thrombospondin motifs 7- (ADAMTS7) Gen ist mit der Koronaren Herzkrankheit assoziiert. Diese Arbeit umfasst eine funktionelle und phänotypische Charakterisierung ADAMTS-7-defizienter humaner glatter Gefäßmuskelzellen, Untersuchungen zur ADAMTS7-Expressionsregulation und eine Evaluation spezifischer Antikörper. ADAMTS-7-Defizienz verminderte die Zellmigration unter proatherogenen Bedingungen nicht. Oxidiertes Low-density lipoprotein bewirkte eine Reduktion der ADAMTS7-Expression, welche durch Knockdown des Oxidized low-density lipoprotein receptor 1 teilweise reversibel war., The A disintegrin and metalloproteinase with thrombospondin motifs 7 (ADAMTS7) gene is associated with coronary artery disease. This work comprises a functional and phenotypical characterization of ADAMTS-7 deficient human vascular smooth muscle cells, analyses of ADAMTS7 gene expression regulation as well as an evaluation of specific antibodies. ADAMTS-7 deficiency did not impair cell migration under proatherogenic conditions. ADAMTS7 expression was reduced by oxidized Low-density lipoprotein, which was partly reversible by Oxidized low-density lipoprotein receptor 1 knockdown.

Highlights • This current review covers studies that have identified long non-coding RNAs in aortic aneurysm development and progression. • We separately discuss transcripts and mechanisms of importance to thoracic as well as abdominal aortic aneurysms. • Functional data on lncRNAs being identified are highlighted. • Some have been studied in human as well as experimental models of the disease pathology. Abstract Aortic aneurysm (AA) is a complex and dangerous vascular disease, featuring progressive and irreversible vessel dilatation. AA is typically detected either by screening, or identified incidentally through imaging studies. To date, no effective pharmacological therapies have been identified for clinical AA management, and either endovascular repair or open surgery remains the only option capable of preventing aneurysm rupture. In recent years, multiple research groups have endeavored to both identify noncoding RNAs and to clarify their function in vascular diseases, including aneurysmal pathologies. Notably, the molecular roles of noncoding RNAs in AA development appear to vary significantly between thoracic aortic aneurysms (TAAs) and abdominal aortic aneurysms (AAAs). Some microRNAs (miRNA - a non-coding RNA subspecies) appear to contribute to AA pathophysiology, with some showing major potential for use as biomarkers or as therapeutic targets. Studies of long noncoding RNAs (lncRNAs) are more limited, and their specific contributions to disease development and progression largely remain unexplored. This review aims to summarize and discuss the most current data on lncRNAs and their mediation of AA pathophysiology.

Aims: Long non-coding RNAs (lncRNAs) have been shown to regulate numerous processes in the human genome, but the function of these transcripts in vascular aging is largely unknown. We aim to characterize the expression of lncRNAs in endothelial aging and analyse the function of the highly conserved lncRNA H19. Methods and results: H19 was downregulated in endothelium of aged mice. In human, atherosclerotic plaques H19 was mainly expressed by endothelial cells and H19 was significantly reduced in comparison to healthy carotid artery biopsies. Loss of H19 led to an upregulation of p16 and p21, reduced proliferation and increased senescence in vitro. Depletion of H19 in aortic rings of young mice inhibited sprouting capacity. We generated endothelial-specific inducible H19 deficient mice (H19iEC-KO), resulting in increased systolic blood pressure compared with control littermates (Ctrl). These H19iEC-KO and Ctrl mice were subjected to hindlimb ischaemia, which showed reduced capillary density in H19iEC-KO mice. Mechanistically, exon array analysis revealed an involvement of H19 in IL-6 signalling. Accordingly, intercellular adhesion molecule 1 and vascular cell adhesion molecule 1 were upregulated upon H19 depletion. A luciferase reporter screen for differential transcription factor activity revealed STAT3 as being induced upon H19 depletion and repressed after H19 overexpression. Furthermore, depletion of H19 increased the phosphorylation of STAT3 at TYR705 and pharmacological inhibition of STAT3 activation abolished the effects of H19 silencing on p21 and vascular cell adhesion molecule 1 expression as well as proliferation. Conclusion: These data reveal a pivotal role for the lncRNA H19 in controlling endothelial cell aging.

Background: The angiogenic function of endothelial cells is regulated by numerous mechanisms, but the impact of long noncoding RNAs (lncRNAs) has hardly been studied. We set out to identify novel and functionally important endothelial lncRNAs. Methods: Epigenetically controlled lncRNAs in human umbilical vein endothelial cells were searched by exon-array analysis after knockdown of the histone demethylase JARID1B. Molecular mechanisms were investigated by RNA pulldown and immunoprecipitation, mass spectrometry, microarray, several knockdown approaches, CRISPR-Cas9, assay for transposase-accessible chromatin sequencing, and chromatin immunoprecipitation in human umbilical vein endothelial cells. Patient samples from lung and tumors were studied for MANTIS expression. Results: A search for epigenetically controlled endothelial lncRNAs yielded lncRNA n342419, here termed MANTIS, as the most strongly regulated lncRNA. Controlled by the histone demethylase JARID1B, MANTIS was downregulated in patients with idiopathic pulmonary arterial hypertension and in rats treated with monocrotaline, whereas it was upregulated in carotid arteries of Macaca fascicularis subjected to atherosclerosis regression diet, and in endothelial cells isolated from human glioblastoma patients. CRISPR/Cas9-mediated deletion or silencing of MANTIS with small interfering RNAs or GapmeRs inhibited angiogenic sprouting and alignment of endothelial cells in response to shear stress. Mechanistically, the nuclear-localized MANTIS lncRNA interacted with BRG1, the catalytic subunit of the switch/sucrose nonfermentable chromatin-remodeling complex. This interaction was required for nucleosome remodeling by keeping the ATPase function of BRG1 active. Thereby, the transcription of key endothelial genes such as SOX18, SMAD6, and COUP-TFII was regulated by ensuring efficient RNA polymerase II machinery binding. Conclusion: MANTIS is a differentially regulated novel lncRNA facilitating endothelial angiogenic fun