The 12-lead electrocardiogram (ECG) recorded during ventricular tachycardia (VT) remains a simple, universally available tool for VT localization. In idiopathic VT, QRS morphology and polarity often reflect the site of earliest activation and can facilitate targeted mapping. In patients with structural heart disease, however, the surface ECG predominantly represents the VT exit region rather than the critical diastolic isthmus; thus, its main clinical value lies in regionalization and in planning ablation strategy and access. A pragmatic stepwise interpretation integrating V1 morphology, precordial transition (V1–V6), and the frontal plane axis can localize VT in most cases, although accuracy is influenced by scar size, conduction abnormalities, antiarrhythmics, and variable exit sites. Emphasis is placed on estimating epicardial or intramural involvement using interval- and morphology-based criteria—especially in nonischemic cardiomyopathy—and on integrating ECG findings with cardiac scar imaging. Key disease-specific ECG patterns are highlighted. Electrocardiographic imaging, automated ECG analysis, and artificial intelligence (AI)-based localization approaches may further improve VT localization.

Vascular aging, endothelial dysfunction, and cellular senescence are interconnected processes that drive cardiovascular diseases (CVDs). Several studies have confirmed that microRNAs (miRNAs) are closely associated with the development of CVDs. Among them, miR-21-5p and miR-146a-5p have emerged as critical players in CVDs: MiR-21-5p is implicated in promoting inflammation and fibrosis, while miR-146a-5p influences vascular smooth muscle cell proliferation and inflammatory responses. Targeting these miRNAs for therapeutic intervention could provide a novel strategy to mitigate the deleterious effects of vascular aging. Recent advances in RNA-based therapeutics have led to the exploration of circular RNAs (circRNAs) as stable and effective miRNA inhibitors. Due to their circular structure, circRNAs exhibit greater stability than linear miRNA inhibitors, such as anti-miRs. This study aimed to evaluate the expression of miR-21-5p and miR-146a-5p in senescent human vascular endothelial cells and vascular smooth muscle cells (VSMCs) and to assess whether synthetic circRNAs could effectively inhibit these miRNAs. In a series of experiments, human umbilical vein endothelial cells (HUVECs) and VSMCs were transfected with custom-designed circRNAs targeting these miRNAs. Here, we want to focus on VSMCs. The effects on miRNA expression, target gene regulation, and cellular functions such as proliferation and migration were analyzed. Our findings demonstrate that circRNA-mediated inhibition of miR-21-5p and miR-146a-5p significantly affects gene expression and cellular behavior in senescent vascular cells, suggesting that circRNAs may serve as more efficient and stable therapeutic agents compared to traditional anti-miRs. By addressing these objectives, this study contributes to the growing body of knowledge on circRNA-mediated miRNA inhibition and its potential implications for vascular health. The results from this study not only enhance our understanding of miR-21-5p and miR-146a-5p in vascular senescence but also provide a foundation for future therapeutic strategies.

Senescent endothelial cells (ECs), characterized by a reduced angiogenic and regenerative potential, are key players in the pathophysiology of cardiovascular diseases. Therefore, targeting these cells has been suggested as an effective therapeutic strategy to increase health. Here, we are the first to report that non-genetic overexpression of the Yamanaka factors induces partial functional rejuvenation and attenuation of senescence-associated features in endothelial cells. Methods to characterize the effects of the transient reprogramming included quantification of gene expression as well as measurements of cellular functions in vitro. Further, in vivo experiments were performed in a hind-limb ischemia model. The application of the pharmacological cocktail to replicative senescent ECs resulted in a robust but timely restricted activation of Oct3/4, Sox2, Klf4, and c-Myc (p < 0.0 and p < 0.01). This was associated with a significant reduction of senescence markers such as p16ink4a and p14arf (p < 0.01). Additionally, qPCR-based telomere length measurements were stabilized, and functional properties of senescent ECs, such as proliferation, migration, sprouting, and tube formation, were improved (p < 0.05). Continuous cultivation of the treated cells over the long term indicated that expression of p16ink4a and p14arf remained significantly low, while cell migration remained enhanced. In vivo, a significantly improved blood flow was observed at 7 and 14 days after hind-limb ischemia in 21 months old C57BL/6 mice (p < 0.001). In conclusion, we revealed that a partial attenuation of endothelial cell senescence-associated features can be induced by a short pharmacological overexpression of the Yamanaka factors. While the compounds used are individually approved for other indications, their combined use in this context highlights a conceptual translational potential, whereas the clinical applicability of this approach remains to be evaluated.

Subjective and objective markers are important in describing healthy aging, yet little is known about their relationships. This study analysed the time-dependent association of dehydroepiandrosterone sulfate (DHEAS) with subjective health. At baseline, DHEAS was measured in participants aged 45-83 randomly selected from the general population. Subjective mental and physical health were assessed using the 12-item Short Form (SF-12) questionnaire at baseline and two follow-ups. In sex-specific linear regression models controlled for age, weight, tobacco consumption, Charlson Comorbidity Index, depression, and testosterone levels, the associations of DHEAS with the subscores of the SF-12 were analysed. DHEAS showed a positive cross-sectional association with subjective physical health, which was stronger in women and remained relevant after multivariable adjustment. However, longitudinal analyses revealed no long-term effect of DHEAS on subjective health. These findings suggest that the association between DHEAS and subjective physical health is temporary and that an underlying causality is unlikely.

Bei einer kardiologischen Nachsorge, drei Monate nach transfemoraler Transkatheteraortenklappenimplantation (TAVI), berichtete eine 83-jährige Patientin über eine neue, einseitige Claudicatio-intermittens-Symptomatik, die die postoperative Wiederaufnahme ihres Nordic Walkings vereitelte. Duplexsonographisch lag eine Malposition des ankerbasierten vaskulären MANTA-Verschlusssystems (VCD) mit Thrombosierung und filiformer Abgangsstenose der A. femoralis superficialis (AFS) vor, hämodynamisch Rutherford I/1 entsprechend. Patientenwunsch und hoher Leidensdruck begründeten eine operative Revaskularisation. Intraoperativ bestätigte sich der subtotale Verschluss der AFS durch Verhaken des intravaskulären Ankers in der Femoralisbifurkation bei zu weit distal eingebrachtem VCD. Im postoperativen Verlauf konnte die Patientin bei Beschwerdefreiheit ihre Nordic-Walking-Aktivitäten wieder aufnehmen.

BACKGROUND:Multivessel coronary artery disease (CAD) is present in 30% to 70% of patients presenting with non-ST-segment elevation myocardial infarction (NSTEMI) depending on varying age and risk profiles. In contrast to the STEMI cohort, there is only limited scientific evidence derived from randomized controlled trials directing the general decision for or against complete revascularization in the NSTEMI population. PRIMARY HYPOTHESIS:The COMPLETE-NSTEMI trial aims to investigate whether multivessel percutaneous coronary intervention (PCI) is superior over culprit-lesion only PCI in patients with NSTEMI and multivessel CAD. DESIGN:COMPLETE-NSTEMI is a prospective, randomized, controlled, multicenter, parallel group, open-label trial. It will enroll 3390 NSTEMI patients with multivessel CAD at 65 to 70 sites in Germany and Austria. Patients will be randomized 1:1 to either complete revascularization with PCI or culprit lesion-only PCI. ENDPOINTS:The primary efficacy endpoint is a composite of cardiovascular death or rehospitalization for nonfatal myocardial infarction during follow-up. The trial is event-driven and will be stopped as soon as 578 primary endpoint events and a minimal follow-up duration of 12 months for each patient are reached. CURRENT STATUS:The first patient was enrolled at October 27, 2023. By April 2025, 51 sites have been activated and >500 patients have been randomized. Completion of recruitment is expected for the first half of 2027. The final results of the primary endpoint are expected in 2028. OUTLOOK:COMPLETE NSTEMI will be the first dedicated trial to answer the question about the optimal revascularization strategy in patients with NSTEMI and multivessel CAD. TRIAL REGISTRATION:CLINICALTRIALS.GOV: NCT05786131.

In der gebietsübergreifenden Intensivmedizin und klinischen Notfallversorgung stellt die Innere Medizin mit ihren Schwerpunkten eine zentrale Säule dar. Dieses Curriculum – zusammengestellt von Mitgliedern der internistischen Fachgesellschaften (DGIIN, DGIM samt Schwerpunktgesellschaften), dem Berufsverband Deutscher Internistinnen und Internisten (BDI) sowie der Deutschen Gesellschaft für Palliativmedizin (DGP) – gibt einen umfassenden Überblick über internistische Weiterbildungsinhalte der Intensiv- und klinischen Notfallmedizin – Kenntnisse, praktische Fähigkeiten (Kompetenzgrad I–III), beruflich-professionelles Verhalten –, deren Erwerb für eine bestmögliche Versorgung der Patienten aus internistischer Sicht erforderlich scheint. Das Curriculum beschreibt zum einen die allgemeinen Aspekte der Internistischen Intensiv- und Klinischen Notfallmedizin mit den Inhalten: Struktur- und Prozessqualität, Notaufnahme: Erstdiagnostik, Initialtherapie und Indikationsstellung zur weiterführenden Behandlung, Schockraumversorgung, intensivmedizinische Syndrome, Diagnostik und Monitoring, generelle Therapieverfahren, Ethik, Hygienemaßnahmen und Pharmakotherapie. Anschließend folgen spezielle Aspekte der Internistischen Intensiv- und Klinischen Notfallmedizin: angiologische/gefäßmedizinische, endokrinologische, diabetologische und metabolische, gastroenterologische und hepatologische, geriatrische, hämatologische und medizinisch-onkologische, infektiologische, kardiologische, nephrologische, palliativmedizinische, pneumologische, rheumatologische und toxikologische. Unterlegt sind die Themen jeweils mit Verweisen auf praxisrelevante Publikationen. Primär für Internisten gedacht zeigt das Curriculum aber auch allen nichtinternistischen Intensiv- und Notfallmedizinern, mit welchen internistischen Krankheitsbildern und Komorbiditäten sie bei ihrer Tätigkeit rechnen müssen.

Objectives. Transcoronary ablation of septal hypertrophy (TASH) and surgical myectomy are the recommended treatment options for patients with hypertrophic obstructive cardiomyopathy refractory (HOCM) when conventional drug treatment is not sufficient. We describe the application of radiofrequency (RF) energy via coronary guidewires in an animal model for selective occlusion of coronary side branches that mimics the principle of TASH. Methods. Transcoronary guidewire ablation of coronary vessels was performed in 5 adult pigs under general anesthesia in an animal cathlab after successful bench testing of the ablation settings. After assessing transcoronary pacing parameters, RF energy was delivered via coronary guidewires insulated by coating or by a monorail balloon and positioned in different coronary side branches. Occlusion or patency of the specific coronary side branch was documented by coronary angiography after RF delivery. Results. After the transcoronary RF ablation, the intended occlusion of the coronary vessel (thrombolysis in myocardial infarction [TIMI]-0 or TIMI-1 flow) could be verified by angiography in 82% of the septal branches attempted and 79% of the epicardial branches. The mean ablation duration was 10.3 seconds at 20-W power with an initial impedance of 176 +/- 31 ohm. No unintended occlusion of the main vessels occurred in any case. Conclusions. RF ablation via coronary guidewires is a feasible method for inducing an acute occlusion of coronary vessels and may change the interventional therapy of HOCM if the current limitations of this technique are overcome.

BACKGROUND:Allostatic load (AL) is a surrogate of the physiological response to stress and reflects the 'wear and tear' on the body. Previous studies indicated that socioeconomic and behavioral determinants influence AL, which in turn is associated with health outcomes. Therefore, AL is increasingly used to operationalize the relationship between social inequality, stress, and health outcomes. This study aimed to investigate associated factors and patterns of AL in the population over a 20-year period using data from the CARLA cohort. METHODS:The analysis included 473 participants from the CARLA study (Cardiovascular Disease, Living and Ageing in Halle), aged 45-80 years at baseline. From recruitment in 2002 in Halle (Saale), three follow-up examinations took place until 2022. We calculated AL scores as the sum of standardized z-scores for metabolic, immune, cardiovascular, and anthropometric components. Descriptive statistics of AL scores were stratified by sex and age categories. Multiple regression analyses were conducted for the first and third follow-up to assess if there were changes in associations between sociodemographic factors and AL. RESULTS:Average AL scores of men decreased, while women's AL scores returned to baseline levels after an initial decrease observed at the first follow-up. Stratified analyses of AL scores revealed that women in the younger age cohorts had lower mean AL scores at baseline than men (women: -3.47, 95% CI [-4.24; -2.71] vs. men: -1.13, 95% CI [-1.84; -0.42] at age <55). At the same time, women showed higher mean AL scores than men in older age cohorts (women: -0.32, 95% CI [-1.58; 0.95] vs. men: -0.93, 95% CI [-1.99; 0.14] at age 65-<70). Results of multiple regression models indicated lower AL scores for women (β: -1.21, 95% CI [-1.93, -0.49]). Professional status was associated with lower AL scores for men but not for women (β: -1.06, 95% CI [-2.02, -0.11] for men). Further, physical activity was negatively associated with AL scores for the total study sample and for women (β: -0.54, 95% CI [-0.82, -0.26]) for total sample and β: -0.74, 95% CI [-1.17, -0.32] for women). CONCLUSION:Our results highlight the importance of health awareness and physical activity for overall health, assessed by AL. Distinct AL score changes and sex-specific socioeconomic influences offer insights into sex-related patterns of aging. Further research is needed to understand the underlying mechanisms of socioeconomic influences on stress-related aging processes between sexes.

Aim: Following myocardial infarction (MI), endothelial cells (ECs) and fibroblasts are key to tissue repair. Modulating their function could improve healing and cardiac recovery. Given limited effective therapies, microRNAs (miRNAs) are promising targets. This project investigates the effects of miR-762 on endothelial and fibroblast function to explore its therapeutic potential in post-MI repair. Methods: Expression of miR-762 was analyzed in vivo after ultrasound-guided MI in C57BL/6J mice by qRT-PCR. Further, miR-762 expression was assessed in human ECs and Aortic Fibroblasts (AoF). The impact of anti-miR-762 transfection on migration, proliferation, angiogenesis, and cell death was evaluated. Potential miR-762 downstream targets were identified by RNA sequencing and validated by qRT-PCR and western blot analysis. Results: MiR-762 is significantly upregulated in vivo 24h after MI ( p <0.05), with no regulation at 72h. Similarly, in vitro, hypoxia led to increased miR-762 levels in ECs at 24h (p<0.05), but not at 72h. Functional analyses indicate that miR-762 does not affect EC proliferation. However, its downregulation significantly enhances EC migration after 6h ( p <0.0001), reduces apoptosis ( p <0.05), and improves the angiogenic capacity ( p <0.05). RNA sequencing revealed that anti-miR transfection significantly downregulates CMPK2 ( p <0.0001) and XAF1 ( p <0.05) in ECs, these findings were confirmed by qRT-PCR. Western blot analyses showed a significant reduction of CMPK2 and XAF1 protein levels ( p <0.05). Upstream pathway analysis revealed a significant downregulation of IFNGR1 and STAT1 proteins ( p <0.05), although no changes were observed at the RNA level. Novel expression analyses revealed significantly higher miR-762 levels in AoF compared to ECs ( p <0.05). Preliminary data suggest that miR-762 downregulation in AoF does not affect proliferation but appears to reduce migration capacity and increase cell death. Conclusion: In summary, our findings identify miR-762 as a promising therapeutic target for enhancing recovery after MI. Inhibition of miR-762 improves EC function and promotes angiogenesis by inhibiting XAF1 and CMPK2, acting through the IFNγ-JAK-STAT signaling pathway. Additionally, miR-762 may modulate AoF activity, potentially leading to decreased scar formation. To further investigate this aspect, additional experiments focusing on AoF are needed. The next phase of our research will explore these mechanisms and include the first in vivo therapeutic studies.

As part of a cardiological follow-up 3 months after transcatheter aortic valve repair (TAVR), an 83-year-old women reported that the attempt to resume postoperative Nordic walking failed due to new unilateral intermittent claudication. Duplex sonography revealed a filiform stenosis at the origin of the superficial femoral artery (SFA), caused by malposition of a plug-based vascular closure device (VCD), hemodynamically consistent with Rutherford I/1. The patient's wishes and high level of suffering justified surgical revascularization. Intraoperatively, subtotal occlusion of the SFA was confirmed by wedging the intravascular anchor of a MANTA VCD in the femoral bifurcation. During the postoperative course, the patient was able to resume her Nordic walking activities without any symptoms.

Senescent endothelial cells (EC) are key players in the pathophysiology of cardiovascular diseases and are characterized by a reduced angiogenic and regenerative potential. Therefore, targeting these cells has been suggested as an effective therapeutic strategy to reduce vascular disease burden and potentially improve health and lifespan of humans. Here, we aimed to establish a pharmacological, partial reprogramming strategy to improve replicative senescent endothelial cell function in the context of angiogenesis. We demonstrate that our treatment improves tube formation and sprouting capacity but also increases proliferation and migration capacity in vitro. Further, inflammation and DNA damage were reduced in the replicative senescent cells. These processes were initiated by a short and timely-restricted overexpression of the Yamanaka-factors induced by our pharmacological strategy. The advantage of these compounds is that they are FDA approved in their respective concentrations which could pave the way for use in a clinical setting.

OBJECTIVES:Protection against severe course of SARS-CoV-2 infection after COVID-19 vaccination or infection was extensively studied. It is unknown whether this effect also translates into shortened duration of mild infections. We assessed the duration of symptoms depending on vaccination status and previous SARS-CoV-2 infections among individuals with a mild course of infection. METHODS:For two post-pandemic winters (2022/2023 and 2023/2024), in total 13,615 participants of the German DigiHero study reported their SARS-CoV-2 infections from September to March. Via negative binomial regression adjusting for sociodemographic factors, we studied the association of infection duration (days with symptoms and in bed) with number of vaccinations, prior SARS-CoV-2 infections, and time since last vaccination/and infection. RESULTS:We noted no major differences in infection duration depending on the number of vaccinations and time since last infection for short mild infections (≤21 days with symptoms). Per 6 months since the last vaccination, symptom duration and days spent in bed increased by 2 % and 4 %. The risk of long mild SARS-CoV-2 infections (>21 days with symptoms) was higher for individuals with no prior SARS-CoV-2 infection (Odds Ratio: 1.98; 95 % confidence interval [1.43; 2.76]), but not for vaccinations (OR: 0.98; 95 % CI [0.74; 1.33]). CONCLUSIONS:There was no indication of reduced duration of symptoms during short mild infections depending on the number of vaccinations and time since the last SARS-CoV-2 vaccination or infection. A prior SARS-CoV-2 infection was protective against prolonged disease in mild SARS-CoV-2 infections.

Herz-Kreislauf-Erkrankungen sind voraussichtlich bis 2050 die Krankheitsbilder mit der höchsten Morbidität und Mortalität und stellen die Versorger vor neue Herausforderungen. Auf diesem Hintergrund wird der teilstationären Versorgung eine zunehmende Bedeutung zukommen. Die Mitglieder der Ordinarienkonferenz Kardiologie e. V. haben sich 2023 und 2024 in Arbeitstreffen mit der zukünftigen Ausrichtung von universitären Kliniken für Kardiologie befasst. Hierauf aufbauend wurde ein Konsensuspapier verfasst, das die strategischen Eckpfeiler, Spezifika und Herausforderungen der universitären Kardiologie zusammenfasst und sich an die Vorstände der Universitätsklinika, den Verband der Universitätsklinika Deutschlands e. V. (VUD), den Gemeinsamen Bundesausschuss (G-BA), Kostenträger und die Entscheidungsträger der Ministerien für Gesundheit, Wissenschaft und Finanzen der jeweiligen Bundesländer sowie im Bund richtet. Die Inhalte dieses Konsensuspapiers werden im vorliegenden Beitrag skizziert.

Coronary artery disease, especially in the elderly population, can lead to calcification of the arteries, which is tightly connected to an unfavorable prognosis. We investigated the combination of coronary intravascular lithotripsy (IVL) to ease deliverability of drug-coated balloons (DCBs) with the potential favorable effect of DCB in avoiding vessel thrombosis due to lack of foreign body implantation. Seventy-three calcified lesions were studied in an open, observational, prospective, single-arm consecutive registry. The role of IVL was studied as to the facilitation of DCB delivery, procedural, angiographic, and clinical outcomes. DCB only angioplasty without stenting was possible in 55 (75

Internal medicine and its associated subspecialities represent an important cornerstone of intensive care and clinical emergency medicine. This curriculum-compiled by members of the German Society of Medical Intensive Care and Emergency Medicine (Deutsche Gesellschaft für Internistische Intensivmedizin und Notfallmedizin), the German Society of Internal Medicine (Deutsche Gesellschaft für Innere Medizin) including subspeciality societies, the Professional Association of German Internists (Berufsverband Deutscher Internistinnen und Internisten, BDI) and the German Association for Palliative Medicine (Deutsche Gesellschaft für Palliativmedizin, DGP)-presents an overview of knowledge, skills (competence levels I-III), behaviors, and attitudes necessary for the highest treatment quality for the internal medicine aspects of intensive care and emergency medicine. It includes general aspects of intensive care and clinical emergency medicine (structure and process quality, emergency department: primary diagnostics and treatment as well as the indication for subsequent treatment, resuscitation room management, clinical syndromes in intensive care medicine, diagnostics and monitoring, general therapeutic measures, ethics, hygiene measures, and pharmacotherapy). Subsequently, specific aspects concerning angiology/vascular medicine, endocrinology, diabetology and metabolism, gastroenterology and hepatology, geriatric medicine, hematology and medical oncology, infectiology, cardiology, nephrology, palliative care, pneumology, rheumatology, and toxicology are addressed. Publications focusing on the content of advanced training are quoted to support this concept. The curriculum is written primarily for internists but may also show practicing intensivists and emergency physicians the broad spectrum of internal medicine diseases and comorbidities presented by patients admitted to the intensive care unit or the emergency department.

Endothelial cell (EC) senescence has been identified as a significant contributor to the development of cardiovascular disease, resulting in impaired angiogenesis and tissue regeneration. Consequently, the reversal of EC senescence emerges as a promising therapeutic strategy to restore vascular function and promote healthy aging. In this study, we demonstrate that pharmacological, non-genetic partial reprogramming via transient activation of the Yamanaka factors Oct3/4, Sox2, Klf4, and c-Myc (OSKM) can rejuvenate senescent ECs. Gene expression profiling and functional assays were performed in vitro, and the regenerative capacity of the murine hind-limb ischemia model was assessed using young and aged C57BL/6J mice. Treatment with the reprogramming cocktail resulted in a robust, transient induction of OSKM at both the mRNA and protein levels (p<0.0001) in replicatively senescent ECs. This was accompanied by a marked reduction in senescence-associated markers, including p16Ink4a, p14Arf, TNFα, IL-1β, IL-6, and CD44 (p<0.01 to p<0.0001) and restoration of EC function. Specifically, the treated cells demonstrated enhanced migration (p<0.001), proliferation (p<0.01), sprouting (p<0.0001), and tube formation (p<0.01). Telomere length was shown to be stabilized (p<0.05) and there was a decrease in reactive oxygen species levels (p<0.05). Furthermore, non-senescent ECs demonstrated no response to the treatment (p>0.05). Notably, following treatment, senescent ECs exhibited diminished expression of senescence markers and maintained their migratory capacity also over time. In vivo, the aged (22-month-old) mice demonstrated significantly enhanced perfusion recovery at 7 and 14 days following the induction of ischaemia (p<0.001), accompanied by an augmentation in capillary density within the gastrocnemius muscle (CD31-staining, p<0.05). Conversely, no effects were observed in young mice (3 months old) (p>0.05). In conclusion, the transient pharmacological induction of OSKM has been demonstrated to reverse endothelial senescence and restore regenerative function in vitro and in vivo. This approach offers a novel and clinically relevant strategy to enhance vascular repair and combat age-related ischemic diseases.

Background: Ageing is a major driving factor for vascular changes. Over time, senescent endothelial and smooth muscle cells accumulate in the vasculature, promoting endothelial dysfunction and cardiovascular disease (CVD). MicroRNAs serve a key function in CVD and cellular senescence. Methods: Expression levels of microRNA-375-3p were analyzed in non-senescent and replicative senescent human coronary artery endothelial cells (HCAEC) and human coronary artery smooth muscle cells (HCASMC) by qRT-PCR. To evaluate its effects regarding cellular functions and cellular senescence, miR-375-3p was downregulated by transfection of a LNA-inhibitor. In silico research identified targets of miR-375-3p, which were validated on mRNA and protein level. Results: In a machine-learning based analysis of over 1200 microRNA datasets including 1800 distinct miRNAs from various tissue samples from patients between the age of 20 to 90 years, we showed that miR-375-3p is an important miR upregulated in aged individuals showing an exceptionally strong centrality and targeting more than 80% of the respective age-associated targets . In vitro, expression analyses showed especially a 10-fold upregulation of miR-375-3p in senescent compared to non-senescent HCAEC (p<0.05). In aortic endothelial cells of mice, miR-375-3p is upregulated in old compared to young mice. Transfection with a LNA-inhibitor downregulated the microRNA successfully. By the inhibition of miR-375-3 we observed improved angiogenic capacities in non-senescent and replicative senescent HCAEC by tube formation and sprouting assays (p<0.05). However, cell death in non-senescent cells was elevated by inhibition. In morphology analyses, downregulation of the miR led to an increase in cell nuclei size in non-senescent cells. Inhibition induced the expression of pro-inflammatory cytokines like TNF-alpha (p<0.01) and IL-1beta (p<0.05) in senescent HCAEC. Conclusion: A machine-learning based analysis identified microRNA-375-3p as a significantly upregulated microRNA in senescent endothelial cells, which was verified in vitro and in vivo. Inhibition of miR-375-3p improved angiogenic capacities in non-senescent and replicative senescent HCAEC. However, its role in cellular senescence remains inconclusive and needs further evaluation.

Atherosclerosis is a primary contributor to cardiovascular mortality. The pathological proliferation, migration to the intima, and phenotypic switching of Smooth Muscle Cells (SMCs) leads to vascular remodeling and plaque development. MicroRNAs (miRs) have emerged as a promising target for modulating SMC activity while maintaining the proper function of Endothelial Cells (ECs). This study investigates whether modulating miR-331-3p can therapeutically target SMCs in atherosclerosis and how this affects ECs. MiR-331-3p levels in SMCs, ECs, and murine tissue were measured via qRT-PCR. Pre- and anti-miR transfection was used to assess its upregulation or inhibition on proliferation, migration, metabolism, and apoptosis. Morphological changes were analyzed by immunofluorescence microscopy. In silico predicted targets were validated at the mRNA level, and protein-level validation is ongoing. Initial investigations revealed a significant increase in miR-331-3p expression during disease progression (p<0.0001) in ApoE--/- mice, a commonly used atherosclerosis model. Further research uncovered that miR-331-3p is highly conserved across species, suggesting its relevance for subsequent in vitro experiments with human SMCs and ECs. Cytokines such as IFNγ, IL-1β, and TNFα were found to influence miR-331-3p expression in SMCs. Following pre- or anti-miRNA transfection, the cellular morphology of SMCs and ECs remained unchanged. Microscopically, the increase in SMC cell area observed 24 hours after miR-331-3p overexpression (p<0.0277) was linked to enhanced proliferation, as validated by BrdU assay results (p<0.05). In contrast, inhibition of miR-331-3p inhibited SMC migration (p<0.0178) and revealed divergent effects on SMC and EC function. Cell death assays showed that miR-331-3p causes anti-apoptotic effects in SMCs 48 hours after regulation. In silico analyses, utilizing literature research, target prediction tools, and public sequencing data, identified several potential targets, including KLF16, BAK1, PHLPP1, SOCS1, TNFα, TGFBR1, and DUSP5. Their upregulation in SMCs was confirmed 24 and 48 hours after miR-331-3p overexpression (p<0.05), while ECs showed remarkably few, if any, target changes. In conclusion, inhibiting miR-331-3p efficiently modulates vascular remodeling in SMCs without impairing EC function, highlighting its potential as a valuable target for cardiovascular treatment strategies potentially by majorly regulating the identified targets DUSP5 and PHLPP1.

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Martin-Luther-Universität Halle-Wittenberg Background and Purpose The accumulation of senescent cells in the vasculature contributes to impaired vascular function over the lifetime. Micro RNAs (miRs) are promising therapeutic targets due to their cell-specific functions. Targeting miR-127-3p emerges as an attractive approach to enhance vascular healing and limit angioplasty-related complications. Here, we focus on elucidating the role of miR-127-3p in vascular smooth muscle cells (VSMC) to characterize their role in cellular function and vascular senescence. Methods Utilizing reverse transcription and qRT-PCR we checked expression levels of miR-127-3p in human and murine aortic VSMCs. After overexpression and downregulation via transfection of pre-miR and antagomiR, we evaluated the influence of miR-127-3p on cellular functions such as proliferation and migration of human (non-) replicative senescent VSMCs. Potential targets were selected via target prediction analysis and verified on mRNA and protein levels. Morphological changes were visualized via high-resolution fluorescence microscopy. Senescence marker expression was checked after transfection by qRT-PCR. Results MiR-127-3p is highly upregulated in femoral artery tissue on day 10 after wire-induced injury in C57Bl6 mice, serving as restenosis model (p<0.001) and aged C57BL/6 mice (3 months vs. 22 months, p<0.0001). Replicative senescent human VSMC exhibit increased miR-127-3p expression (p<0.05). In vitro, miR-127-3p overexpression reduces proliferation in senescent (p<0.001) and non-senescent (p<0.01) cells. It hinders migration in non-senescent cells (p<0.0001) but has a reverse effect in senescent cells (p<0.01). Potential targets affecting cellular functions, inflammatory pathways, and cellular aging were identified as mTOR, AXL, and TEAD3 (p<0.05). Transfection of pre-miR leads to increase in cell size and the acquisition of a flattened morphology of human VSMC. P14arf and laminB1, as senescence-associated genes, showed downregulation after overexpression of miR-127-3p on mRNA level (p<0.05). When VSMCs were stimulated with anti-miR-127-3p, increased mRNA levels of p14arf and laminB1 were detected (p<0.05). Conclusion This study identifies miR-127-3p as a contributor to cellular functions including cellular proliferation and migration and seems to have opposing effects in non- and replicative senescent VSMC. Further research is needed to validate miR-127-3p as a therapeutic target for vascular aging and remodeling in vitro and in vivo.