Observational studies suggest an association between impaired oral health and cardiovascular disease; however, the directionality and underlying mechanisms remain unclear. In particular, whether heart failure (HF) itself adversely affects oral and periodontal health has not been systematically investigated in large populations or experimental models.We examined the association between HF and self-reported oral health indicators in 502,387 participants of the UK Biobank, including 17,356 individuals with HF defined by ICD-9/10 codes. Multivariable logistic regression models adjusted for demographic factors, cardiovascular comorbidities, systemic inflammation, lifestyle, and socioeconomic status were applied. To explore causality and mechanisms, periodontal tissue remodeling and inflammation were assessed in a murine model of pressure overload-induced HF using transverse aortic constriction (TAC). Periodontal ligament (PDL) space and alveolar bone microarchitecture were quantified by micro-computed tomography, and gingival inflammatory gene expression was analyzed by RT-PCR.HF patients exhibited a significantly higher prevalence of oral health burden compared with controls (51% vs. 40%, p<0.001). HF was associated with a 1.6-fold increased risk of impaired oral health, which remained significant after full adjustment (adjusted OR 1.18, 95% CI 1.14-1.22; p<0.001). In mice, reduced left ventricular ejection fraction following TAC was strongly associated with expansion of the maxillary PDL space (R2 = 0.63, p = 0.009) and alterations in alveolar bone microarchitecture (trabecular thickness R2 = 0.41 p = 0.061, trabecular number R2 = 0.38 p = 0.07). These structural changes were accompanied by increased gingival expression of pro-inflammatory cytokines, including Il1b (SHAM vs. TAC: 1.44 ± 1.02 vs. 3.39 ± 1.53, p = 0.06) and TNF-α (SHAM vs. TAC: 1.37 ± 0.86 vs. 5.71 ± 1.23, p = 0.002 predominantly in the maxilla.HF is independently associated with impaired oral health in a large population cohort and induces site-specific periodontal inflammation and remodelling in experimental HF. These findings support HF as an upstream driver of compromised oral-periodontal health, challenging the prevailing concept that oral disease primarily contributes to cardiovascular pathology.

AIM:Cardiovascular diseases are the leading cause of death worldwide, with arterial calcification being a risk factor, especially in patients with diabetes and kidney disease. Arterial calcification involves hydroxyapatite deposition and the transformation of vascular smooth muscle cells (vSMCs) into osteoblast-like cells, processes mediated in part by tissue-nonspecific alkaline phosphatase (TNAP, encoded by ALPL), a key regulator of mineralization. Previous work showed increased activity of electron transport chain complexes I and IV during vSMC calcification. This study examined the role of mitochondrial complex I in vSMC calcification. METHODS:vSMCs were calcified using osteogenic medium. Rotenone was used as a complex I inhibitor. Metabolomic profiling, extracellular pH measurements, and TNAP activity assays were performed. RESULTS:Rotenone dose-dependently reduced matrix mineralization, with near-complete inhibition at 50 nM, without affecting cell viability. Metabolomic analysis showed that rotenone increased both intracellular and extracellular lactic acid while decreasing pyruvic acid, indicating a shift toward glycolysis. Exogenous lactic acid reduced mineralization by 24%; sodium lactate reduced mineralization without altering extracellular pH; and extracellular acidification independently reduced mineralization by 23%, indicating that both lactate and proton-mediated acidification partially contribute to the anti-calcific mechanism. Additionally, rotenone decreased TNAP activity by 59% without affecting ALPL transcript levels. CONCLUSION:Inhibition of mitochondrial complex I causes metabolic reprogramming in calcifying vSMCs, promoting lactic acid accumulation that partially inhibits matrix mineralization through lactate-mediated and acidification-dependent mechanisms. The near-complete inhibition of calcification by rotenone suggests additional uncharacterized mechanisms beyond the lactic acid-pH axis contribute to its full anti-calcific effect, warranting further investigation.

Background The aryl hydrocarbon receptor (AhR) is a well-described regulator of xenobiotic stimuli, but recent studies highlight a potential bidirectional role in mediating lipid metabolism. The precise mechanisms underlying these effects are not yet fully understood. Therefore, this study aimed to investigate the impact of a genetic aberration in the AhR in hepatocytes and intestinal epithelial cells on lipid metabolism and the development of atherosclerosis. Methods By injecting Albumin-iCre containing AAVs into PCSK9 overexpressing Ahrfl/fl mice, hepatocyte-specific Ahr-deficient mice were created. Additionally, mice with an intestinal epithelial cell-specific deficiency of the Ahr (Ahrfl/fl VillinCre+) and control mice (Ahrfl/fl) were injected with AAV8-PCSK9. Both mouse models were fed a high-fat diet (HFD) for 12 weeks, after which alterations in plasma lipid levels and the development of atherosclerosis were assessed. Findings Mice lacking hepatocyte-specific Ahr did not exhibit significant differences in lipid levels or atherosclerosis. In contrast, male mice lacking Ahr in intestinal epithelial cells (IECs) showed significant changes in intestinal and systemic lipid levels, which coincided with altered RNA transcription and intestinal kinase activity, whereas female mice were unaffected. Interestingly, female mice showed reduced circulating leukocyte counts, whereas males were unaffected. Combined, these alterations resulted in decreased atherosclerosis development in both sexes. Interpretation This study reveals a major impact of intestinal epithelial cell-specific Ahr on lipid metabolism, with effects on atherosclerosis development, particularly in male mice, whereas atherosclerosis development in female intestinal epithelial cell-specific Ahr-deficient mice is accompanied by reduced circulating leukocytes, thereby suggesting apparent sex dimorphism.

AIMS:Patients with chronic kidney disease (CKD) display a reduced survival following myocardial infarction (MI). As the underlying mechanisms remain unclear, we examined the impact of CKD on cardiac remodeling and function post-MI using a mouse model of adenine-induced CKD. METHODS AND RESULTS:After MI, CKD mice showed a stronger cardiac dysfunction compared to non-CKD controls. While immunohistochemical and immunofluorescence analyses did not reveal changes in cardiomyocyte apoptosis, infarction size, or myofibroblast content, CKD mice exhibited an increased number of circulating myeloid cells post-infarction and more neutrophil infiltration in the heart. Combining RNAseq, untargeted kinome profiling, western blotting, and mass spectrometry revealed that post-MI, CKD enhanced cardiac oxidative stress and the acute stress complex S100A8/A9 in circulation and the heart, and enforced cardiac MAP-kinase p38 activation and NR4A1 phosphorylation as pathways underlying cardiomyocyte dysfunction. S100A8/A9 also exerted an acute detrimental impact on calcium flux and sarcomere shortening in cardiomyocytes ex vivo. Increased myeloid cell-derived S100A8/A9 expression was confirmed in the infarcted human heart by single-nucleus RNAseq, and CKD patients had higher post-infarction S100A8/A9 levels compared to patients without kidney dysfunction. Furthermore, integrating metabolomics, RNAseq, and mitochondrial analysis uncovered a disturbed cardiac metabolism with impaired glycolysis, a reduced glycerol-3-phosphate-shuttle, and a reduced Coenzyme A-bioavailability in CKD vs. non-CKD mice post-MI. These alterations were associated with poorer cardiac performance post-MI, without intrinsic defects in mitochondrial function observed. CONCLUSION:Our study reveals innate immune activation, inflammation, oxidative stress, and metabolic alterations indicative of reduced glycolytic entry and CoA bioavailability along with aggravated cardiac dysfunction post-MI in CKD compared to non-CKD conditions, independent of infarct size, and with poorer cardiac performance in CKD associated with the cardiac metabolic alterations. Combined, this could contribute to the worsened outcome of CKD patients post-MI and reveals cardiac metabolism in CKD as an interesting translational research target.

Abstract Background Myeloproliferative neoplasms (MPN) comprise a heterogenous group of hematological malignancies that include polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF). Current therapeutic strategies rely on cytoreductive approaches that mitigate disease burden and thromboembolic risk but are not curative. Allogeneic stem cell transplantation remains the only curative option, underscoring the need for novel therapeutic strategies. We previously identified hypoxia–inducible factor 1 (HIF-‍‍1) as a selective vulnerability in JAK2V617F–positive cells, but the underlying metabolic mechanisms remain incompletely defined. Methods In vitro studies utilized 32D cells transduced with an empty vector control, Jak2WT, or Jak2V617F. To evaluate metabolic dependencies, CRISPR-Cas9 was used to generate Slc2a1 (GLUT1) and Slc2a3 (GLUT3) knockout clones, which were subsequently characterized via RNA sequencing, extracellular flux analysis, and cellular fitness assays (proliferation, viability, and apoptosis). Pharmacological targeted inhibition of GLUT1/3 was evaluated in human JAK2V617F–mutated post-MPN AML cell lines (SET-2, HEL), primary patient-derived cells and a Jak2V617F knock-in mouse model. Combinatorial efficacy was assessed using the JAK1/2 inhibitor ruxolitinib. Results JAK2V617F induced HIF-1–dependent metabolic reprogramming, characterized by increased glycolytic flux and oxidative metabolism. Complete abrogation of glucose uptake occurred only upon combined loss of GLUT1 and GLUT3 in Jak2V617F cells, revealing functional redundancy between these transporters that sustains enhanced glycolysis. Disruption of glucose uptake selectively induced stress–associated transcriptional programs and replication stress, triggering an S-phase arrest that culminated in apoptosis and impaired viability, specifically in Jak2V617F cells. In vivo, pharmacological inhibition of HIF-1 or GLUT induced a reorganization of erythropoiesis to the spleen but did not ameliorate core disease features. In contrast, in vitro GLUT inhibition robustly reduced cell viability in human SET-2 and HEL cell lines and impaired proliferation, viability, and colony formation in patient-derived PBMCs. Conclusions Collectively, these findings establish HIF-1–driven glucose metabolism as a metabolic vulnerability in JAK2V617F-positive MPN. The selective exhaustion of patient–derived clones defines the HIF-1–GLUT1/3 axis as a central, targetable bottleneck. These data provide a mechanistic rationale for further investigation of HIF-1 or GLUT inhibitors, suggesting that targeting this fundamental requirement may help overcome clinical limitations to achieve disease modification and eradicate the malignant clone.

Patients with chronic kidney disease are at an increased risk of developing heart failure, but the underlying mechanisms remain incompletely understood, at least in part because of the paucity of mouse models of uremic cardiomyopathy. In this study, we used two different experimental setups of 2,8-dihydroxyadenine-induced nephropathy in different mouse strains to develop a non-invasive mouse model of uremic cardiomyopathy. Among the different models, only 129/Sv mice fed an adenine-supplemented diet for 16 weeks showed typical features of uremic cardiomyopathy. Kidney damage was confirmed by histopathologic findings of diffuse fibrosis with collagen deposition, crystal formation, and uremia. The cardiac phenotype showed significantly increased myocardial fibrosis associated with impaired cardiac contractility under dobutamine-induced stress conditions. This was associated with a significant activation of the mTOR pathway and downstream endoplasmic reticulum stress, increased apoptosis, and inflammation. Treatment of 129/Sv mice with an adenine-supplemented diet for 16 weeks represents a model of uremic cardiomyopathy with increased myocardial fibrosis and impaired cardiac function, as well as a shift from cardioprotective to detrimental signaling, increased endoplasmic reticulum stress, and inflammation.

BACKGROUND AND AIMS:Bioactive adrenomedullin 1-52 (bio-ADM) is a novel biomarker for the assessment of endothelial function and prediction of adverse outcomes in patients with acute heart failure and cardiogenic shock. The SMART (Second Manifestations of Arterial Disease) risk score is a validated tool for risk assessment in patients with established atherosclerotic cardiovascular disease (ASCVD). Here we assessed whether bio-ADM adds incremental prognostic value to the SMART risk score in stable patients with ASCVD. METHODS:Circulating bio-ADM levels were measured in 452 stable patients with ASCVD. Endpoints evaluated were all-cause and cardiovascular mortality; follow up was 3 years. RESULTS:Bio-ADM was higher in non-survivors (n = 45; median 36.8 pg/mL) compared to survivors (n = 407; median 18.3 pg/mL; p < 0.0001). Bio-ADM was found to be a strong predictor for all-cause mortality (Chi2: 44.58; C-index: 0.79) as well as cardiovascular death (Chi2: 33.29; C-index: 0.85) and proved to be superior to other markers including hs-Troponin T (Chi2: 7.77; C-index: 0.73) and eGFRCKD-EPI 2021 (Chi2: 25.10; C-index: 0.70). In multivariable analyses adjusting for age, sex, diabetes mellitus, hypertension, smoking, NT-proBNP, and eGFRCKD-EPI 2021, bio-ADM remained independently associated with all-cause mortality (HR: 1.6; 95 % CI: 1.2-2.1; Chi2: 96.17; p < 0.00001; C-index: 0.89) and cardiovascular death (HR: 1.7; 95 % CI: 1.1-2.5; Chi2: 57.71; p < 0.00001; C-index: 0.88). Addition of bio-ADM to the SMART risk score meaningfully improved model performance in predicting mortality (SMART risk score: Chi2: 19.91; p = 0.0001; C-index: 0.69; SMART risk score + bio-ADM: Chi2: 54.51; p < 0.00001; C-index: 0.81). CONCLUSIONS:Bio-ADM levels are independently associated with mortality and provide incremental added value on top of the SMART risk score in stable patients with ASCVD.

Glucagon-like peptide-1 (GLP-1) is a gut-derived peptide secreted in response to nutritional and inflammatory stimuli. Elevated GLP-1 levels predict adverse outcome in patients with acute myocardial infarction or sepsis. GLP-1 holds cardioprotective effects and GLP-1 receptor agonists reduce cardiovascular events in high-risk patients with diabetes. In this study, we aimed to investigate the capacity of GLP-1 to predict outcome in patients with cardiogenic shock (CS) complicating myocardial infarction. Circulating GLP-1 levels were serially assessed in 172 individuals during index PCI and day 2 in a prospectively planned biomarker substudy of the IABP-SHOCK II trial. All-cause mortality at short- (30 days), intermediate- (1 year), and long-term (6 years) follow-up was used for outcome assessment. Patients with fatal short-term outcome (n = 70) exhibited higher GLP-1 levels [86 (interquartile range 45–130) pM] at ICU admission in comparison to patients with 30-day survival [48 (interquartile range 33–78) pM; p < 0.001] (n = 102). Repeated measures ANOVA revealed a significant interaction of GLP-1 dynamics from baseline to day 2 between survivors and non-survivors (p = 0.04). GLP-1 levels above vs. below the median proved to be predictive for short- [hazard ratio (HR) 2.43; 95 www.ClinicalTrials.gov Identifier: NCT00491036.

Background Proenkephalin A 119-159 (penKid) is a novel blood biomarker for real-time assessment of kidney function and was found to be independently associated with worsening kidney function and mortality. A novel penKid-based estimated glomerular filtration rate equation (eGFR(PENK-Crea)), outperforms current creatinine-based eGFR equations in predicting iohexol or iothalamate plasma clearance-based measured GFR. In this study, we aimed to evaluate the predictive value of penKid and eGFR(PENK-Crea) for all-cause mortality in stable patients at high cardiovascular risk. Methods Circulating penKid levels were assessed in 615 stable patients hospitalized at the Department of Cardiology at University Hospital Aachen, Germany. The endpoint was all-cause mortality; follow up was 3 years. Results penKid levels were higher in 46 non-survivors [58.8 (IQR 47.5-85.0) pmol/l] compared to 569 survivors [43.8 (IQR 34.0-58.0) pmol/l; P < .0001]. Univariable Cox regression analyses found penKid and eGFR(PENK-Crea) to be associated with all-cause mortality (C index 0.703, chi(2) 33.27, P < .00001; C index 0.716, chi(2) 36.51, P < .00001). This association remained significant after adjustment for significant baseline parameters including age, smoking, chronic heart failure, use of diuretics, leucocytes, body mass index, sex, and creatinine (C index 0.799, chi(2) 72.06, P < .00001). Importantly, penKid provided significant added value on top of eGFR(CKD-EPI 2021) (eGFR(CKD-EPI 2021): C index 0.716, chi(2) 34.21; eGFR(CKD-EPI 2021 )+ penKid: C index 0.727, chi(2): 40.02; Delta chi(2) 5.81; all P < .00001) for all-cause mortality prediction in our cohort. Conclusions penKid levels and eGFR(PENK-Crea) is associated with all-cause mortality within a 3-year follow-up period and the addition of penKid on top of eGFR(CKD-EPI 2021) provided significant added value in mortality prediction.

Chronic kidney disease (CKD) significantly increases cardiovascular risk and mortality, and the accumulation of uremic toxins in the circulation upon kidney failure contributes to this increased risk. We thus performed a screening for potential novel mediators of reduced cardiovascular health starting from dialysate obtained after hemodialysis of patients with CKD. The dialysate was gradually fractionated to increased purity using orthogonal chromatography steps, with each fraction screened for a potential negative impact on the metabolic activity of cardiomyocytes using a high-throughput MTT-assay, until ultimately a highly purified fraction with strong effects on cardiomyocyte health was retained. Mass spectrometry and nuclear magnetic resonance identified the metabolite mycophenolic acid-β-glucuronide (MPA-G) as a responsible substance. MPA-G is the main metabolite from the immunosuppressive agent MPA that is supplied in the form of mycophenolate mofetil (MMF) to patients in preparation for and after transplantation or for treatment of autoimmune and non-transplant kidney diseases. The adverse effect of MPA-G on cardiomyocytes was confirmed in vitro, reducing the overall metabolic activity and cellular respiration while increasing mitochondrial reactive oxygen species production in cardiomyocytes at concentrations detected in MMF-treated patients with failing kidney function. This study draws attention to the potential adverse effects of long-term high MMF dosing, specifically in patients with severely reduced kidney function already displaying a highly increased cardiovascular risk.

Abstract Background Bioactive adrenomedullin 1-52 (bio-ADM) is a dynamic blood biomarker for real-time assessment of endothelial function. Bio-ADM was recently shown to be a prognostic marker in patients with acute heart failure and cardiogenic shock. The SMART (Second Manifestations of Arterial Disease) score is a validated tool for risk assessment in patients with established atherosclerotic cardiovascular disease (ASCVD). The aim of this study was to assess whether measurement of bio-ADM adds incremental value to the SMART score in stable patients with ASCVD. Methods Circulating bio-ADM levels were assessed in n=695 stable patients with ASCVD in an all-comer cohort. Endpoints evaluated were all-cause mortality and cardiovascular mortality; follow up was 3 years. Results Bio-ADM was higher in non-survivors (all-cause death: n=54, median 33.1 pg/mL) compared to survivors (n=641, median 17.9 pg/mL; p<0.0001). Univariable Cox regression analyses showed bio-ADM to be associated with adverse outcome [standardized hazard ratio (HR) of bio-ADM values: All-cause death: 2.4, 95% confidence interval (CI): 2.0-2.9; p<0.001, cardiovascular death: 2.5, 95% CI: 1.9-3.3; p<0.001]. This association remained significant in various multivariable Cox regression models. Bio-ADM was found to be a strong marker for mortality (c-index: 0.80, Chi²: 54.1) and proved to be superior to other markers including hs-Troponin T (c-index: 0.61, Chi²: 2.0) and eGFR CKD-EPI 2021 (c-index: 0.687, Chi²: 33.5). Addition of bio-ADM to the SMART score significantly improved model performance in predicting mortality (SMART score: c-index: 0.717, Chi²: 24.73; SMART score + bio-ADM: c-index: 0.832, Chi²: 63.24; Delta c-index: 0.115; Delta Chi²: 38.51; all p<0.001) and showed net reclassification improvement in risk categorization with 11.6% in the group of survivors and 9.8% for non-survivors. Conclusion Bio-ADM provides incremental added value (improved discrimination, calibration and reclassification) on top of the SMART risk score in patients with ASCVD.

BACKGROUND:Cardiac hypertrophy is characterized by remodeling of the myocardium, which involves alterations in the ECM (extracellular matrix) and cardiomyocyte structure. These alterations critically contribute to impaired contractility and relaxation, ultimately leading to heart failure. Emerging evidence implicates that extracellular signaling molecules are critically involved in the pathogenesis of cardiac hypertrophy and remodeling. The immunophilin CyPA (cyclophilin A) has been identified as a potential culprit. In this study, we aimed to unravel the interplay between eCyPA (extracellular CyPA) and myocardial dysfunction and evaluate the therapeutic potential of inhibiting its extracellular accumulation to improve heart function.METHODS:Employing a multidisciplinary approach encompassing in silico, in vitro, in vivo, and ex vivo experiments we studied a mouse model of cardiac hypertrophy and human heart specimen to decipher the interaction of CyPA and the cardiac microenvironment in highly relevant pre-/clinical settings. Myocardial expression of CyPA (immunohistology) and the inflammatory transcriptome (NanoString) was analyzed in human cardiac tissue derived from patients with nonischemic, noninflammatory congestive heart failure (n=187). These analyses were paralleled by a mouse model of Ang (angiotensin) II-induced heart failure, which was assessed by functional (echocardiography), structural (immunohistology, atomic force microscopy), and biomolecular (Raman spectroscopy) analyses. The effect of inhibiting eCyPA in the cardiac microenvironment was evaluated using a newly developed neutralizing anti-eCyPA monoclonal antibody.RESULTS:We observed a significant accumulation of eCyPA in both human and murine-failing hearts. Importantly, higher eCyPA expression was associated with poor clinical outcomes in patients (P=0.043) and contractile dysfunction in mice (Pearson correlation coefficient, -0.73). Further, myocardial expression of eCyPA was critically associated with an increase in myocardial hypertrophy, inflammation, fibrosis, stiffness, and cardiac dysfunction in vivo. Antibody-based inhibition of eCyPA prevented (Ang II)-induced myocardial remodeling and dysfunction in mice.CONCLUSIONS:Our study provides strong evidence of the pathogenic role of eCyPA in remodeling, myocardial stiffening, and dysfunction in heart failure. The findings suggest that antibody-based inhibition of eCyPA may offer a novel therapeutic strategy for nonischemic heart failure. Further research is needed to evaluate the translational potential of these interventions in human patients with cardiac hypertrophy.

Aim: Chronic kidney disease (CKD) is accompanied by increased cardiovascular risk and heart failure (HF). In rodents, 2,8-dihydroxyadenine (DHA)-induced nephropathy is a frequently used CKD model. Cardiac and kidney tubular cells share high energy demand to guarantee constant contractive force of the heart or reabsorption/ secretion of primary filtrated molecules and waste products by the kidney. Here we analyze time-dependent mechanisms of kidney damage and cardiac consequences under consideration of energetic pathways with the focus on mitochondrial function and lipid metabolism in mice.Methods and results: CKD was induced by alternating dietary adenine supplementation (0.2 % or 0.05 % of adenine) in C57BL/6J mice for 9 weeks. Progressive kidney damage led to reduced creatinine clearance, kidney fibrosis and renal inflammation after 3, 6, and 9 weeks. No difference in cardiac function, mitochondrial respiration nor left ventricular fibrosis was observed at any time point. Investigating mechanisms of renal damage, protective SirT3 was decreased in CKD, which contrasted an increase in protein kinase B (AKT) expression, mechanistic target of rapamycin (mTOR) downstream signaling, induction of oxidative and endo-plasmic reticulum (ER) stress. This occurred together with impaired renal mitochondrial function and accu-mulation of hexosylceramides (HexCer) as an established mediator of inflammation and mitochondrial dysfunction in the kidney.Conclusions: 2,8-DHA-induced CKD results in renal activation of the mTOR downstream signaling, endoplasmic reticulum stress, tubular injury, fibrosis, inflammation, oxidative stress and impaired kidney mitochondrial function in conjunction with renal hexosylceramide accumulation in C57BL/6J mice.

Abstract Background GLP-1 and GLP-2 (glucagon-like peptide-1/2) are incretin hormones that are co-secreted from intestinal L-cells in response to food intake and inflammatory stimuli. While GLP-1 is known to induce postprandial insulin secretion and to improve cardiovascular outcomes in patients with diabetes, GLP-2 enhances intestinal nutrient absorption. GLP-2 agonists are clinically used for the treatment of patients with short bowel syndrome. The relevance of GLP-2 beyond the gut is not well understood. The aim of this study was to investigate the role of GLP-2 for cardiovascular disease (CVD). Methods Total GLP-2 levels were assessed at time of admission in 918 patients with myocardial infarction presenting with acute chest pain, among them 597 patients with NSTEMI and 321 with STEMI. The primary composite outcome of the study was the first occurrence of all-cause death, nonfatal myocardial infarction, or nonfatal stroke (3-P-MACE) with a median follow-up of 311 days. To induce atherosclerosis, Glp2r−/− or WT mice were injected with PCSK9 virus and fed a diet high in cholesterol (HCD) for 12 weeks. Results Kaplan-Meier survival plots (separated by the median of GLP-2 with a cut-off value of 4.4 pM) and univariable cox regression analyses found GLP-2 values to be associated with adverse outcome (3-P-MACE and all-cause mortality; logarithmized GLP-2 values HR: 2.87; p<.0001). Further adjustment for age, sex, smoking, hypertension, hypercholesterinemia, previous CVD and diabetes mellitus did not affect this association (logarithmized GLP-2 values HR: 2.66; p=0.0055). Receiver operating characteristic curve (ROC) analyses illustrated that GLP-2 is a strong indicator for early events (area under the curve of the combined endpoint at 7 days: 0.74; 14 days: 0.76; 30 days: 0.76; 6 months: 0.72), which proved to be superior to Troponin T and hs-CRP. To asses the functional role of GLP-2 in CVD in an experimental approach we injected Glp2r−/− or WT mice PCSK9 virus (to induce functional Ldlr-deficiency and hypercholesterolemia) and fed these mice a HCD. After 12 weeks Glp2r−/− mice compared to WT littermates presented with a significant reduction in plaque volume and lesion size. While body weight and circulating leukocyte numbers (FACS analysis) were unaffected, Glp2r−/− mice had lower cholesterol levels. Conclusion Circulating GLP-2 levels are associated with cardiovascular events in patients with acute myocardial infarction while inactivation of the GLP-2 system reduces atherosclerosis in mice. Future studies are needed to investigate whether the GLP-2 receptor might provide a novel therapeutic target for cardiovascular disease.

Metabolic derangement is a key culprit in kidney pathophysiology. Organoids have emerged as a promising in vitro tool for kidney research. Here, we present a fine-tuned protocol to analyze bioenergetics in single human induced-pluripotent-stem-cell (iPSC)-derived kidney organoids using Seahorse XF96. We describe the generation of self-organized three-dimensional kidney or-ganoids, followed by preparation of organoids for Seahorse XF96 analysis. We then detail how to carry out stress tests to determine mitochondrial and glyco-lytic rates in single kidney organoids.

Background: GLP-1 and GLP-2 (glucagon-like peptide-1/2) are incretin hormones that are co-secreted from intestinal L-cells in response to food. While GLP-1 is known to induce postprandial insulin secretion and to improve cardiovascular outcomes in patients with diabetes, GLP-2 is a local intestinal growth factor enhancing intestinal nutrient absorption. GLP-2 agonists are clinically used for the treatment of patients with short bowel syndrome. The relevance of GLP-2 beyond the gut is not well understood. The aim of this study was to investigate the role of GLP-2 for cardiovascular disease (CVD). Methods: Circulating GLP-2 levels were assessed at time of hospital admission in 1929 patients with myocardial infarction. The primary outcome of the study was the first occurrence of cardiovascular death, nonfatal myocardial infarction, or nonfatal stroke (3-P-MACE). To induce atherosclerosis, Glp2r –/– or WT mice were injected with PCSK9 virus and fed a HCD for 12 weeks. Results: Kaplan-Meier survival plots and univariable cox regression analyses found GLP-2 to be associated with adverse outcome (3-P-MACE and all-cause mortality; log. GLP-2 values HR: 2.87; p<.0001). Further adjustment for age, sex, smoking, hypertension, hypercholesterinemia, previous CVD and diabetes mellitus did not affect this association (log. GLP-2 values HR: 2.66; p=0.0055). Receiver operating characteristic curve (ROC) analyses illustrated that GLP-2 is a strong indicator for early events, which proved to be superior to Troponin T and hs-CRP. To asses the functional role of GLP-2 in CVD in an experimental approach we injected Glp2r –/– or WT mice PCSK9 virus and fed these mice a HCD. After 12 weeks Glp2r –/– mice compared to WT littermates presented with a significant reduction in plaque volume and lesion size. While body weight and leukocyte numbers in various organs were unaffected, Glp2r –/– mice had significantly lower cholesterol levels. Conclusion: Circulating GLP-2 levels are independently associated with cardiovascular events in patients with myocardial infarction while inactivation of the GLP-2 system reduces atherosclerosis in mice. Future studies are needed to investigate whether the GLP-2 receptor might provide a novel therapeutic target for cardiovascular disease.

Cardiac remodeling occurs frequently in chronic kidney disease patients and affects quality of life and survival. Current treatment options are highly inadequate. As kidney function declines, numerous metabolic pathways are disturbed. Kidney and heart functions are highly connected by organ crosstalk. Among others, altered volume and pressure status, ischemia, accelerated atherosclerosis and arteriosclerosis, disturbed mineral metabolism, renal anemia, activation of the renin-angiotensin system, uremic toxins, oxidative stress and upregulation of cytokines stress the sensitive interplay between different cardiac cell types. The fatal consequences are left-ventricular hypertrophy, fibrosis and capillary rarefaction, which lead to systolic and/or diastolic left-ventricular failure. Furthermore, fibrosis triggers electric instability and sudden cardiac death. This review focuses on established and potential pathophysiological cardiorenal crosstalk mechanisms that drive uremia-induced senescence and disease progression, including potential known targets and animal models that might help us to better understand the disease and to identify novel therapeutics.

Abstract Aims In this prospective, placebo‐controlled, double‐blind, exploratory study, we examined early and more delayed effects of empagliflozin treatment on haemodynamic parameters (primary endpoint: cardiac output) and kidney function including parameters of acute kidney injury (AKI) in patients with acute decompensated heart failure (HF). Methods and results Patients with acute decompensated HF with or without diabetes were randomized to empagliflozin 10 mg or placebo for 30 days. Haemodynamic, laboratory, and urinary parameters were assessed after 6 h, 1 day, 3 days, 7 days, and 30 days of treatment. Median time between hospital admission and randomization was 72 h. Baseline characteristics were not different in the empagliflozin (n = 10) and placebo (n = 9) groups. Empagliflozin led to a significant increase in urinary glucose excretion throughout the study (baseline: 37 ± 15 mg/24 h; Day 1: 14 565 ± 8663 mg/24 h; P = 0.001). Empagliflozin did not affect the primary endpoint of cardiac index or on systemic vascular resistance index at any time point. However, empagliflozin significantly reduced parameters of AKI (urinary TIMP‐2 and IGFBP7 by NephroCheck® as indicators of tubular kidney damage), which became significant after 3 days of treatment [placebo: 1.1 ± 1.1 (ng/mL)2/1000; empagliflozin: 0.3 ± 0.2 (ng/mL)2/1000; P = 0.02] and remained significant at the 7 day time point [placebo: 2.5 ± 3.8 (ng/mL)2/1000; empagliflozin: 0.3 ± 0.2 (ng/mL)2/1000; P = 0.003]. Conclusions In this study, empagliflozin treatment did not affect haemodynamic parameters but significantly reduced markers of tubular injury in patients with acute decompensated HF.