Low extracellular vesicle concentrations predict survival in patients with heart failure.

Background: Heart disease is of worldwide importance due to high morbidity and mortality. Extracellular vesicle (EV) concentration and size represent novel diagnostic and prognostic biomarkers, e.g. in patients with liver cancer, but data on their prognostic relevance in heart disease are lacking. Here, we investigated the role of EV concentration, size and zeta potential in patients with heart disease. Methods: Vesicle size distribution, concentration and zeta potential were measured by nanoparticle tracking analysis (NTA) in 28 intensive care unit (ICU) and 20 standard care (SC) patients and 20 healthy controls. Results: Patients with any disease had a lower zeta potential compared to the healthy controls. Vesicle size (X50) was significantly higher in ICU patients (245 nm) with heart disease as compared to those patients with heart disease receiving standard care (195 nm), or healthy controls (215 nm) (p = 0.001). Notably, EV concentration was lower in ICU patients with heart disease (4.68 × 1010 particles/ml) compared to SC patients with heart disease (7,62 × 1010 particles/ml) and healthy controls (1.50 × 1011 particles/ml) (p = 0.002). Extracellular vesicle concentration is prognostic for overall survival in patients with heart disease. Overall survival is significantly reduced when the vesicle concentration is below 5.55 × 1010 particles/ml. Median overall survival was only 140 days in patients with vesicle concentrations below 5.55 × 1010 particles/ml compared to 211 days in patients with vesicle concentrations above 5.55 × 1010 particles/ml (p = 0.032). Summary: Concentration of EVs is a novel prognostic marker in ICU and SC patients with heart disease.

Dataset of ileum bacterial diversity in mice after heart failure due to pressure overload.

We recently reported the correlation of gut bacterial diversity with heart failure using a mouse model of heart failure due to pressure overload induced by transverse aortic constriction (TAC). We found that gut the bacterial diversity is significantly altered and is directly correlated to the severity of heart failure (Heart Failure Severity Closely Correlates with Intestinal Dysbiosis and Subsequent Metabolomic Alterations (Spehlmann, 2022). In addition, stool samples that were collected for the gut microbial diversity analysis, we dissected ileum from the mice after 42 days of TAC. The total DNA was extracted to identify the bacterial diversity resided in ileum using 16S rRNA gene amplicon shotgun sequencing and downstream bioinformatics analysis to determine if it is correlated to the heart failure.

Heart Failure Severity Closely Correlates with Intestinal Dysbiosis and Subsequent Metabolomic Alterations.

Growing evidence suggests an altered gut microbiome in patients with heart failure (HF). However, the exact interrelationship between microbiota, HF, and its consequences on the metabolome are still unknown. We thus aimed here to decipher the association between the severity and progression of HF and the gut microbiome composition and circulating metabolites. Using a mouse model of transverse aortic constriction (TAC), gut bacterial diversity was found to be significantly lower in mice as early as day 7 post-TAC compared to Sham controls (p = 0.03), with a gradual progressive decrease in alpha-diversity on days 7, 14, and 42 (p = 0.014, p = 0.0016, p = 0.0021) compared to day 0, which coincided with compensated hypertrophy, maladaptive hypertrophy, and overtly failing hearts, respectively. Strikingly, segregated analysis based on the severity of the cardiac dysfunction (EF < 40% vs. EF 40−55%) manifested marked differences in the abundance and the grouping of several taxa. Multivariate analysis of plasma metabolites and bacterial diversity produced a strong correlation of metabolic alterations, such as reduced short-chain fatty acids and an increase in primary bile acids, with a differential abundance of distinct bacteria in HF. In conclusion, we showed that HF begets HF, likely via a vicious cycle of an altered microbiome and metabolic products.

Prevention and treatment of venous thromboembolism in cancer patients.

Deep vein thrombosis and pulmonary artery embolism are common and serious concomitant diseases in patients with cancer. The prophylaxis and therapy of such venous thromboembolic events (VTE) in oncology have so far been achieved with low-molecular-weight heparins. An increasing number of studies show evidence of the use of direct oral anticoagulants. However, since none of the possible options were shown to have a clear advantage in all patients, the individual decision to use a drug should be made depending on its effectiveness in preventing VTE, the risk of bleeding, the nature of the cancer, the interactions with other medications, the route of administration, and finally the cost of treatment.

Identification of Disease-associated Traits and Clonotypes in the T Cell Receptor Repertoire of Monozygotic Twins Affected by Inflammatory Bowel Diseases.

BACKGROUND AND AIMS: Intestinal inflammation in inflammatory bowel diseases [IBD] is thought to be T cell mediated and therefore dependent on the interaction between the T cell receptor [TCR] and human leukocyte antigen [HLA] proteins expressed on antigen presenting cells. The collection of all TCRs in one individual, known as the TCR repertoire, is characterised by enormous diversity and inter-individual variability. It was shown that healthy monozygotic [MZ] twins are more similar in their TCR repertoire than unrelated individuals. Therefore MZ twins, concordant or discordant for IBD, may be useful to identify disease-related and non-genetic factors in the TCR repertoire which could potentially be used as disease biomarkers. METHODS: Employing unique molecular barcoding that can distinguish between polymerase chain reaction [PCR] artefacts and true sequence variation, we performed deep TCRα and TCRß repertoire profiling of the peripheral blood of 28 MZ twin pairs from Denmark and Germany, 24 of whom were discordant and four concordant for IBD. RESULTS: We observed disease- and smoking-associated traits such as sharing, diversity and abundance of specific clonotypes in the TCR repertoire of IBD patients, and particularly in patients with active disease, compared with their healthy twins. CONCLUSIONS: Our findings identified TCR repertoire features specific for smokers and IBD patients, particularly when signs of disease activity were present. These findings are a first step towards the application of TCR repertoire analyses as a valuable tool to characterise inflammatory bowel diseases and to identify potential biomarkers and true disease causes.

Serum Levels of miR-143 Predict Survival in Critically Ill Patients.

BACKGROUND AND AIMS: Recent data suggested a potential role of miR-143 as a biomarker for systemic inflammation and infection. However, its role in critical illness and sepsis is only poorly understood. METHODS: We determined circulating levels of miR-143 in 218 critically ill patients, of which 135 fulfilled sepsis criteria, and compared them to 76 healthy controls. Results were correlated with clinical records. RESULTS: In the total cohort of critically ill patients from a medical intensive care unit (ICU), miR-143 serum levels tended to be lower compared to healthy control samples, but this difference did not reach statistical significance. In ICU patients, serum levels of miR-143 were independent of disease etiology, including the presence of sepsis, or severity of disease. Importantly, low miR-143 serum levels were associated with an unfavorable short- and long-term prognosis in ICU patients. Our study identified different optimal cut-off values at which low miR-143 serum levels predicted mortality with a high diagnostic accuracy. In line with this, concentrations of circulating miR-143 correlated with markers of organ failure such as creatinine, bilirubin, or lactate in our cohort of critically ill patients. CONCLUSION: Low miR-143 serum levels are indicative for an unfavorable short- and long-term prognosis in critically ill patients admitted to a medical ICU. Our data suggest a previously unrecognized role for miR-143 measurements as a novel prognostic marker in critically ill patients.

Serum Levels of Kisspeptin Are Elevated in Patients with Pancreatic Cancer.

Pancreatic adenocarcinoma (PDAC) still represents a devastating disease associated with a very limited survival. Novel biomarkers allowing an early diagnosis as well as an optimal selection of suitable treatment options for individual patients are urgently needed to improve the dismal outcome of PDAC patients. Recently, alterations of Kisspeptin serum levels, a member of the adipokine family, were described in various types of cancers. However, the role of circulating Kisspeptin as a biomarker in PDAC patients is poorly defined. In this study, we measured Kisspeptin serum levels in a cohort of 128 prospectively enrolled PDAC patients undergoing surgical resection as well as 36 healthy controls. Kisspeptin concentrations were elevated in PDAC patients compared to control samples. Nevertheless, Kisspeptin serum levels were independent of tumor-related factors such as the tumor grading, TNM stage, or clinical features such as the ECOG performance status. Finally, in our analysis, neither preoperative nor postoperative Kisspeptin levels turned out as a significant predictor of overall survival after tumor resection. In conclusion, our data suggest that Kisspeptin concentrations are altered in PDAC patients but do not allow to predict patients' outcome after resection of PDAC.

A Combined Score of Circulating miRNAs Allows Outcome Prediction in Critically Ill Patients.

BACKGROUND AND AIMS: Identification of patients with increased risk of mortality represents an important prerequisite for an adapted adequate and individualized treatment of critically ill patients. Circulating micro-RNA (miRNA) levels have been suggested as potential biomarkers at the intensive care unit (ICU), but none of the investigated miRNAs displayed a sufficient sensitivity or specificity to be routinely employed as a single marker in clinical practice. METHODS AND RESULTS: We recently described alterations in serum levels of 7 miRNAs (miR-122, miR-133a, miR-143, miR-150, miR-155, miR-192, and miR-223) in critically ill patients at a medical ICU. In this study, we re-analyzed these previously published data and performed a combined analysis of these markers to unravel their potential as a prognostic scoring system in the context of critical illness. Based on the Youden's index method, cut-off values were systematically defined for dysregulated miRNAs, and a "miRNA survival score" was calculated. Patients with high scores displayed a dramatically impaired prognosis compared to patients with low values. Additionally, the predictive power of our score could be further increased when the patient's age was additionally incorporated into this score. CONCLUSIONS: We describe the first miRNA-based biomarker score for prediction of medical patients' outcome during and after ICU treatment. Adding the patients' age into this score was associated with a further increase in its predictive power. Further studies are needed to validate the clinical utility of this score in risk-stratifying critically ill patients.

miR-155 Predicts Long-Term Mortality in Critically Ill Patients Younger than 65 Years.

INTRODUCTION: Alterations in miR-155 serum levels have been described in inflammatory and infectious diseases. Moreover, a role for miR-155 in aging and age-related diseases was recently suggested. We therefore analyzed a potential age-dependent prognostic value of circulating miR-155 as a serum-based marker in critical illness. METHODS: Concentrations of circulating miR-155 were determined in 218 critically ill patients and 76 healthy controls. RESULTS: By using qPCR, we demonstrate that miR-155 serum levels are elevated in patients with critical illness when compared to controls. Notably, levels of circulating miR-155 were independent on the severity of disease, the disease etiology, or the presence of sepsis. In the total cohort, miR-155 was not an indicator for patient survival. Intriguingly, when patients were subdivided according to their age upon admission to the ICU into those younger than 65 years, lower levels of miR-155 turned out as a strong marker, indicating patient mortality with a similar accuracy than other markers frequently used to evaluate critically ill patients on a medical ICU. CONCLUSION: In summary, the data provided within this study suggest an age-specific role of miR-155 as a prognostic biomarker in patients younger than 65 years. Our study is the first to describe an age-dependent miRNA-based prognostic biomarker in human diseases.

Elevated serum levels of bone sialoprotein (BSP) predict long-term mortality in patients with pancreatic adenocarcinoma.

Patients with pancreatic adenocarcinoma (PDAC) still face a very limited prognosis. At early stage, surgical tumor resection might offer long-term survival but disease recurrence is common and the existing stratification algorithms are often unsuitable to identify patients who particularly benefit from surgery. Here, we investigated the potential role of bone sialoprotein (BSP) as a circulating marker in patients undergoing resection of PDAC. We used ELISA to determine serum concentrations of BSP in a cohort of 132 PDAC patients as well as 39 healthy controls. Circulating BSP levels were significantly higher in PDAC patients compared to healthy controls. Notably, elevated preoperative BSP levels above the ideal cut-off value of 4743 pg/ml turned out as a significant predictor for an impaired postoperative survival. The potential of preoperative BSP levels as a prognostic marker was further underlined by uni- and multivariate Cox-regression analyses including various tumour- and patient-specific. Finally, high tumoral BSP expression was also associated with a significantly impaired long-term survival. In conclusion, we identified a novel role of circulating BSP as a biomarker in PDAC patients undergoing tumor resection. Such data might help to establish new preoperative stratification strategies to better identify patients who particularly benefit from tumor resection.

Serum levels of kisspeptin are elevated in critically ill patients.

INTRODUCTION: Members of the adipokine family such as resistin, adiponectin and omentin have recently been described as novel biomarkers with a diagnostic and prognostic role in the context of critically ill patients during intensive care unit (ICU) treatment. Kisspeptin represent another member of this family and has been shown to be closely correlated to different members of the adipokine family in manifold diseases. However, its role in critical illness and sepsis is currently unknown. MATERIALS AND METHODS: Kisspeptin serum concentrations were measured in 133 ICU patients admitted to the medical ICU. Results were compared with 36 healthy controls. RESULTS: Kisspeptin serum levels were elevated in the serum of critically ill patients at admission to the ICU, when compared to healthy controls, and remained increased after 72 hours of ICU treatment. Notably, kisspeptin levels were independent of the presence of sepsis and etiology of critical illness. In line, serum concentrations of kisspeptin were not correlated to concentrations of inflammatory cytokines or established sepsis markers. Serum kisspeptin correlated inversely with the glomerular filtration rate. In contrast to the reported role of other members of the adipokine family, serum levels of kisspeptin were neither predictive for short term survival during ICU treatment nor for patients' overall survival. Kisspeptin levels did not correlate with other adipokines measured in serum, including leptin, resistin, ghrelin, or adiponectin. CONCLUSIONS: Although circulating kisspeptin levels were strongly elevated in ICU-patients, elevated kisspeptin levels were not predictive for an impaired patients' survival.

Progress in heart failure treatment in Germany.

Acute and chronic heart failure is still a major cause of morbidity and mortality in Europe. Nevertheless, significant progress has been made in diagnosis and treatment of heart failure as well as in unraveling its molecular causes. Here, we focus on some relevant contributions to these achievements by German cardiovascular clinicians and scientists.

Elevated serum levels of bone sialoprotein during ICU treatment predict long-term mortality in critically ill patients.

Bone sialoprotein (BSP), a member of the SIBLINGs (for Small Integrin-Binding LIgand, N-linked Glycoproteins) family, has recently be associated to inflammatory and infectious diseases. We therefore measured BSP concentrations in 136 patients at admission to the intensive care unit (ICU) and 3 days of ICU. BSP levels were compared to 36 healthy blood donors and correlated to clinical data. In these analysis, BSP serum levels were strongly elevated at the time point of admission to the ICU when compared to healthy controls. Moreover BSP concentrations were significantly elevated after 3 days of treatment on the intensive care unit. A further increase in BSP levels was detected in patients with higher APACHE-II-scores and in patients with septic disease. While in most patients, BSP levels decreased during the first three days of treatment on a medical ICU, patients with persistently elevated BSP levels displayed an unfavorable outcome. In these patients, persistently elevated BSP concentrations were a superior predictor of mortality than established indicators of patient´ prognosis such as the SAPS2 or the APACHE-II score. In summary, our data argue for a novel utility for BSP as a biomarker in patients treated on a medical ICU.

Cardio-toxicity of checkpoint inhibitors.

Immune checkpoint inhibitors (ICIs) have started revolutionizing the treatment of numerous advanced oncological diseases by restoring immune resistance against cancer cells. ICI-associated cardiac adverse effects are rare, but severe. About 50% of cardiac complications comprise myocarditis with variable clinical presentation and a high rate of fatality. The pathomechanism is incompletely understood and may involve preexisting autoimmunity such as autoantibodies or common epitopes shared by cardiomyocytes and tumor cells. Especially patients at risk might be followed up by serial troponin measurements in order to allow an early identification of ICI-associated myocarditis. Therapeutic options are limited and consist of early discontinuation of ICI treatment and initiation of an immunosuppression. Further studies are necessary to elucidate the mechanism, define diagnostic criteria, improve surveillance of patients at risk, and finally refine therapy.

Radial access protects from contrast media induced nephropathy after cardiac catheterization procedures.

OBJECTIVES: To assess, whether cardiac catheterization via radial access prevents contrast-induced nephropathy. BACKGROUND: Contrast-induced nephropathy (CIN) is a major clinical problem which accounts for more than 10% of acute kidney injury cases in hospitalized patients. Protective measures such as the infusion of isotonic saline solution or acetylcysteine have not consistently been proven to prevent acute kidney injury (AKI). However, there is growing evidence that radial access for coronary angiography and coronary intervention is associated with a lower incidence of AKI compared to femoral access. METHODS AND RESULTS: In a retrospective monocentric analysis, 2937 patients that had undergone cardiac catheterization were examined. Up to 2013, coronary intervention was performed primarily via the femoral artery in our hospital; thereafter, interventions were primarily done via the radial artery. In the cohort under study, 1141 patients had received catheterization using the radial access while 1796 were examined via the femoral artery. No significant differences were found in the two groups regarding the amount of iodinated contrast medium applied [femoral group: 180 (120-260) ml; radial group: 180 (120-250) ml; P = 0.438]. A total of 400 (13.6%) patients developed acute kidney injury (AKI) after cardiac catheterization (85.3% AKI stage 1; 12.8% AKI stage 2; 2% AKI stage 3). AKI was significantly less frequent in patients that had received radial access compared to patients with femoral access (10.1 vs. 15.9%, P < 0.001). Multivariate regression analysis showed that patient age (1.03/year; 95% CI 1.02-1.04/year; P < 0.001), the amount of contrast media applied (OR 1.003/ml; 95% CI 1.002-1.004/ml; P < 0.001), acute coronary syndrome (OR 2.01, 95% CI 1.52-2.66; P < 0.001), CKD (OR 1.62, 95% CI 1.50-1.70; P < 0.001), pre-existing heart failure (OR 1.27, 95% CI 1.00-1.42 P = 0.007), previous myocardial infarction (OR 1.34, 95% CI 1.15-1.49; P = 0.001), diabetes (OR 1.25, 95% CI 1.04-1.41; P = 0.020) and serum creatinine before the procedure (1.45/mg/dl; 95% CI 1.24-1.69/mg/dl; P < 0.001) were important risk factors for the occurrence of AKI. Our analysis points to a significant risk reduction using radial access (OR 0.65; 95% CI 0.51-0.83; P < 0.001). Interestingly, this reduction in risk was also evident in patients with CKD (OR 0.59; 95% CI 0.41-0.87; P = 0.007). The superiority of radial access was particularly obvious in the subgroup of patients with acute coronary syndrome (13.1% AKI in the radial access group vs. 23.6% AKI in the femoral access group, OR 0.52; 95% CI 0.34-0.81; P = 0.003). CONCLUSION: Our study shows that cardiac catheterization using radial access bears significantly lower risk of AKI than cardiac catheterization via femoral access. The advantage of radial access in acute coronary syndrome regarding morbidity and mortality could partly be explained by the here demonstrated reduced risk for AKI. Thus, radial access should be preferred in patients at risk for AKI.

Absence of RNase H2 triggers generation of immunogenic micronuclei removed by autophagy.

Hypomorphic mutations in the DNA repair enzyme RNase H2 cause the neuroinflammatory autoimmune disorder Aicardi-Goutières syndrome (AGS). Endogenous nucleic acids are believed to accumulate in patient cells and instigate pathogenic type I interferon expression. However, the underlying nucleic acid species amassing in the absence of RNase H2 has not been established yet. Here, we report that murine RNase H2 knockout cells accumulated cytosolic DNA aggregates virtually indistinguishable from micronuclei. RNase H2-dependent micronuclei were surrounded by nuclear lamina and most of them contained damaged DNA. Importantly, they induced expression of interferon-stimulated genes (ISGs) and co-localized with the nucleic acid sensor cGAS. Moreover, micronuclei associated with RNase H2 deficiency were cleared by autophagy. Consequently, induction of autophagy by pharmacological mTOR inhibition resulted in a significant reduction of cytosolic DNA and the accompanied interferon signature. Autophagy induction might therefore represent a viable therapeutic option for RNase H2-dependent disease. Endogenous retroelements have previously been proposed as a source of self-nucleic acids triggering inappropriate activation of the immune system in AGS. We used human RNase H2-knockout cells generated by CRISPR/Cas9 to investigate the impact of RNase H2 on retroelement propagation. Surprisingly, replication of LINE-1 and Alu elements was blunted in cells lacking RNase H2, establishing RNase H2 as essential host factor for the mobilisation of endogenous retrotransposons.

Heart failure is associated with depletion of core intestinal microbiota.

AIMS: In spite of current medical treatment approaches, mortality of chronic heart failure (HF) remains high and novel treatment modalities are thus urgently needed. A recent theory proposes a possible impact of the intestinal microbiome on the incidence and clinical course of heart failure. This study sought to systematically investigate, if there are specific changes of the intestinal microbiome in heart failure patients. METHODS AND RESULTS: The intestinal microbiome of 20 patients with heart failure with reduced ejection fraction due to ischemic or dilated cardiomyopathy was investigated by applying high-throughput sequencing of the bacterial 16S rRNA gene. Microbial profiles were compared to those of matched controls in which heart failure was ruled out by clinical assessment and NT-proBNP serum levels (n = 20). According to the Shannon diversity index (which measures the intra-individual alpha-diversity) based on the distribution of operational taxonomic units (OTUs), HF cases showed a nominally significantly lower diversity index compared to controls (Pnom. = 0.01), and testing for genera abundance showed a tendency towards a decreased alpha diversity of HF patients. Beta-diversity measures (inter-individual diversity) revealed a highly significant separation of HF cases and controls, (e.g. Pweighted UniFracv = 0.004). Assessing the individual abundance of core measurable microbiota (CMM), a significant decrease of Coriobacteriaceae, Erysipelotrichaceae and Ruminococcaceae was observed on the family level. In line with that, Blautia, Collinsella, uncl. Erysipelotrichaceae and uncl. Ruminococcaceae showed a significant decrease in HF cases compared to controls on the genus level. CONCLUSIONS: Heart failure patients showed a significantly decreased diversity of the intestinal microbiome as well as a downregulation of key intestinal bacterial groups. Our data point to an altered intestinal microbiome as a potential player in the pathogenesis and progression of heart failure.

Elevated Omentin Serum Levels Predict Long-Term Survival in Critically Ill Patients.

Introduction. Omentin, a recently described adipokine, was shown to be involved in the pathophysiology of inflammatory and infectious diseases. However, its role in critical illness and sepsis is currently unknown. Materials and Methods. Omentin serum concentrations were measured in 117 ICU-patients (84 with septic and 33 with nonseptic disease etiology) admitted to the medical ICU. Results were compared with 50 healthy controls. Results. Omentin serum levels of critically ill patients at admission to the ICU or after 72 hours of ICU treatment were similar compared to healthy controls. Moreover, circulating omentin levels were independent of sepsis and etiology of critical illness. Notably, serum concentrations of omentin could not be linked to concentrations of inflammatory cytokines or routinely used sepsis markers. While serum levels of omentin were not predictive for short term survival during ICU treatment, low omentin concentrations were an independent predictor of patients' overall survival. Omentin levels strongly correlated with that of other adipokines (e.g., leptin receptor or adiponectin), which have also been identified as prognostic markers in critical illness. Conclusions. Although circulating omentin levels did not differ between ICU-patients and controls, elevated omentin levels were predictive for an impaired patients' long term survival.