Metabolism

BackgroundObesity is a serious risk factor for cardiovascular diseases. A high fat diet results in cellular oxidative stress and endothelial dysfunction in resistance-sized arteries, characterized by reduced nitric oxide (NO) and endothelium-dependent hyperpolarizing (EDH) responses. Thioredoxin-1, a sulfo-oxidoreductase protein that cleaves disulfide bridges between two adjacent cysteine residues in oxidized proteins, has been shown to lower blood pressure and improve endothelium-dependent relaxing responses in aged C57Bl6/J mice.\n\nMethods and ResultsYoung ([~] 3 month-old) male C57Bl6/J mice were fed a high fat diet (42% kcal from fat; obese) or a normal chow (lean) for 3 months. Mice were administered recombinant human thioredoxin-1 (rhTrx; 25 mg/kg) or saline (0.9% NaCl) via tail vein injection at the start, after one month, and after two months. Body weight (BW) was comparable between lean/rhTrx1 and lean/saline at the time of euthanasia (32 {+/-}1 g versus 32 {+/-} 1 g). The high fat regimen resulted in a comparable BW between obese/saline and obese/rhTrx mice (47 {+/-} 1 g versus 45 {+/-} 2 g, respectively). Small (second-order branches) mesenteric arteries (MA2), coronary and femoral arteries were isolated and mounted on the wire-myograph. MA2 and femoral arteries from obese/saline had blunted acetylcholine (10-9 - 10-5 M)-mediated relaxations compared to lean/saline mice, but not to the NO donor sodium nitroprusside. NO and EDH-mediated relaxing responses were blunted in MA2 from obese/lean mice compared to the three other groups.\n\nConclusionTail vein injections with rhTrx prevented endothelial dysfunction in obese mice by improving NO and EDH relaxing responses in MA2.

NAD+ deficiency underlies obesity-induced metabolic disturbances. Here we evaluated the treatment effect of a new and potent NAD+ enhancer, dihydronicotinamide riboside (NRH), in diet-induced obese mice with hyperglycemia and hyperlipidemia. Administering NRH for 7 weeks improved glucose homeostasis by enhancing pancreatic beta-cell functional mass, increasing muscle insulin sensitivity, and reducing hepatic gluconeogenesis. NRH treatment also mobilized fat deposition, reduced circulating lipid, and improved white adipose function. Significant elevation in multi-tissue NAD+ levels and sirtuin (SIRT) activities, especially SIRT3, mediated these metabolic improvements. Inhibiting adenosine kinase (ADK), a newly recognized enzyme in the NRH-induced NAD+ synthesis pathway, blocked NRHs effect in improving glucose and lipid metabolism. ADK inhibition also reduced tissue NAD+ elevation and the subsequent activation of SIRT3, suggesting an active ADK pathway is necessary for NRH-induced metabolic benefits. These observations, for the first time, establish NRH as a promising intervention for correcting obesity-induced metabolic syndrome.

We conducted a comprehensive genetic investigation of obesity in a cohort of 93,673 Korean individuals, categorized by both body mass index and waist circumference using Korean-specific and international criteria. To explore the genetic architecture of obesity and its comorbidities, we performed genome-wide association studies and constructed polygenic risk scores (PRSs) using both conventional single trait and advanced multiple-trait models, including the PRSsum approach. Our analyses identified genome-wide significant loci and demonstrated higher heritability for general obesity than abdominal obesity, and for moderate compared to severe obesity. Notably, East Asian populations showed stronger genetic correlations between abdominal obesity and obesity-related diseases. Both single trait and multiple trait PRSs stratified individuals by risk, with low PRS individuals exhibiting reduced risk for obesity, hypertension, and type 2 diabetes, while high PRS individuals displayed elevated risk, particularly under the multiple trait model. Additionally, interaction and mediation analyses revealed distinct genetic pathways through which obesity contributes to disease development. Collectively, our findings uncover key loci and shared genetic mechanisms linking obesity and its comorbidities in the Korean population. These insights highlight the value of multiple trait PRS models and underscore the importance of ancestry-specific genetic research for addressing the obesity epidemic.

ObjectiveThis research aims to uncover the factors associated with circulating FGF21 levels in a cohort mimicking metabolic disease progression, examining its relationship with adipose tissue (AT) morphology and function. It also investigates FGF21 level changes post-metabolic surgery, predictive factors, and their links to metabolic adjustments. DesignIn this observational study, serum FGF21 was measured in 678 individuals cross-sectionally and longitudinally in 189 undergoing metabolic surgery. We explored links between FGF21, AT histology, cardiometabolic risk factors, weight loss, glucose metabolism changes using feature selection algorithms, univariate/multivariate models, and transcriptome/proteome network analyses in subcutaneous and visceral AT. ResultsFGF21 levels track closely with central adiposity, subclinical inflammation, insulin resistance, and cardiometabolic risk, with circulating leptin emerging as the top predictor. Visceral AT inflammation was associated with liver dysfunction and FGF21 elevation. Post-surgery, FGF21 peaked transitorily at 3 months and predicted fat mass loss at 12 months but not HOMA-IR improvements. Mediation analysis indicated an increased catabolic and AT-lipolytic state associated with higher liver enzyme and FGF21 levels (total effect 0.38, p<0.01; proportion mediation 32%, p<0.01). AT fibrosis was related to a blunted transitory FGF21 increase, and lower fat loss, and hence linked with a reduced surgical effect (FFA and visceral AT fibrosis: rho=-0.31, p=0.030; FFA and fat-mass loss: rho=0.17, p=0.020). ConclusionFGF21 reflects the livers metabolic response to AT characteristics in both central adiposity and after metabolic surgery, with its dynamics reflecting AT-liver crosstalk.

ObjectiveMaternal obesity is usually associated with placental hypovascularity. This study aimed to challenge this paradigm by investigating the immediate vascular adaptations in the placental labyrinth zone in response to short-term, diet-induced obesity. The study hypothesised that there would be an initial compensatory hypervascularisation before the onset of systemic metabolic disease. MethodsFemale Wistar rats were fed either a standard control diet (CG, n = 9) or an ultra-processed, hypercaloric cafeteria diet (EG, n = 9) for eight weeks to induce obesity. On gestational day 16.5, maternal morphometric and biochemical analyses were performed alongside detailed placental histomorphometry and immunohistochemistry for CD31/-actin, in order to quantify foetal vessel density in the labyrinth zone using ImageJ. ResultsThe cafeteria diet successfully induced a significant obese phenotype (mean weight gain: 62.73 g in the experimental group (EG) versus 32.26 g in the control group (CG); P < 0.0001), but did not induce significant hyperglycaemia or dyslipidaemia (P > 0.05). Although there were no significant differences in foetal or placental weights, the labyrinth zone of the EG showed a significant increase in foetal vessel density (29.08 {+/-} 1.91 vessels/field) compared to the CG (26.06 {+/-} 1.80 vessels/field; P = 0.014), indicating robust vascular remodelling. ConclusionShort-term exposure to an obesogenic diet triggers significant compensatory hypervascularisation in the placenta of rats, which is an adaptive response that precedes systemic metabolic dysfunction. This finding contrasts with the hypovascularity observed in chronic obesity. The discrepancy between increased vascular density and foetal growth underscores the importance of evaluating not only the quantity but also the quality and function of blood vessels when assessing placental health in cases of maternal obesity. HighlightsO_LIAcute cafeteria diet-induced obesity causes placental hypervascularity. C_LIO_LIIncreased placental vasculature does not correlate with enhanced foetal growth. C_LIO_LIRapid obesity develops without major pre-gestational metabolic disruption. C_LIO_LIA biphasic model of vascular adaptation to maternal obesity is proposed. C_LI Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=112 SRC="FIGDIR/small/664837v1_ufig1.gif" ALT="Figure 1"> View larger version (43K): org.highwire.dtl.DTLVardef@1090dfaorg.highwire.dtl.DTLVardef@174c7cdorg.highwire.dtl.DTLVardef@1e9a413org.highwire.dtl.DTLVardef@131bf9_HPS_FORMAT_FIGEXP M_FIG C_FIG

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive cancer with poor outcomes. Obesity is a risk factor for several cancers including PDAC due to metabolic dysregulation and inflammation. The ketogenic diet (KD) can alter metabolism and has been evaluated for its effects on tumor progression in non-obese but not obese PDAC using genetically engineered mouse models (GEMMs). We hypothesized that ketone bodies and a KD alter cell and tumor metabolism. We show that ketone treatments altered pyrimidine metabolism in PDAC cells. Moreover, in an obese PDAC GEMM, KD prevented tumor progression independent of weight loss but promoted PDAC in a non-obese PDAC GEMM. The KD-specific delay of obesity-associated PDAC was associated with pancreatic metabolic shifts in pyrimidine, cysteine and methionine, and arginine and proline pathways. These findings suggest potential benefits of a KD in preventing obesity-associated PDAC, but highlights some risks in non-obese settings.

Glucokinase activators (GKAs) have been developed as blood glucose lowering drugs for type 2 diabetes. Despite good short-term efficacy, several GKAs showed a decline in efficacy chronically during clinical trials. The underlying mechanisms remain incompletely understood. We tested the hypothesis that deficiency in the liver glucokinase regulatory protein (GKRP) as occurs with common human GCKR variants affects chronic GKA efficacy. We used a Gckr-P446L mouse model for the GCKR exonic rs1260326 (P446L) variant and the Gckr-del/wt mouse to model transcriptional deficiency to test for chronic efficacy of the GKA, AZD1656 in GKRP-deficient states. In the Gckr-P446L mouse, the blood glucose lowering efficacy of AZD1656 (3 mg/kg body wt) after 2 weeks was independent of genotype. However after 19 weeks, efficacy was maintained in wild-type but declined in the LL genotype, in conjunction with raised hepatic glucokinase activity and without raised liver lipids. Sustained blood glucose lowering efficacy in wild-type mice was associated with qualitatively similar but more modest changes in the liver transcriptome compared with the P446L genotype, consistent with GKA therapy representing a more modest glucokinase excess than the P446L genotype. Chronic treatment with AZD1656 in the Gckr-del/wt mouse was associated with raised liver triglyceride and hepatocyte microvesicular steatosis. The results show that in mouse models of liver GKRP deficiency in conjunction with functional liver glucokinase excess as occurs in association with common human GCKR variants, GKRP-deficiency predisposes to declining efficacy of the GKA in lowering blood glucose and to GKA induced elevation in liver lipids.

Gastric bypass surgery results in long-term weight loss due to re-routing of the gastro-intestinal anatomy and dietary intake alterations. Studies have examined protein change during rapid weight loss (up to 1 year post-surgery), but whether protein changes are maintained long-term after weight stabilization is unknown. To identify proteins and pathways involved with the long-term beneficial effects of weight loss, abundances of 1297 blood-circulating proteins were measured at baseline, 2 and 12 years after Roux-en-Y gastric bypass surgery. Protein changes were compared between 234 surgery and 144 non-surgery subjects with severe obesity, with discovery and replication subgroups. Seventy-one protein changes were associated with 12-year BMI changes and 58 (7 unique) with surgical status. Protein changes, including ApoM, were most strongly associated with long-term changes in lipids (HDL-C and triglycerides). Inflammation, adipogenesis, cellular signaling, and complement pathways were implicated. Short-term improvements in protein levels were maintained long-term, even after some weight regain.

BackgroundEndothelial dysfunction, dyslipidemia, and myocardial injury are major contributors to cardiovascular disease. Avocado (Persea americana), rich in monounsaturated fatty acids and phytochemicals, has shown lipid-lowering and anti-inflammatory properties, but its integrated effects on vascular injury remain unclear. MethodsMale rats were randomized into six groups (n = 4 per group): control, avocado, L-NAME, L-NAME+drugs (metoprolol+losartan), L-NAME+avocado, and L-NAME+drugs+avocado. Morphometric indices, lipid profiles, cardiac injury enzymes, and vascular biomarkers were measured after treatment. One-way ANOVA with Tukey test assessed group differences, while contour plots and correlation networks visualized biomarker interactions. ResultsL-NAME treatment induced a pathological phenotype characterized by reduced feed efficiency (-40%), weight gain (-80%), and BMI (-18%), together with dyslipidemia (LDL +120%, TG +55%, TC +42%, HDL -28%), myocardial stress (troponin +70%, CK +50%, LDH +35%), and vascular activation (endothelin +350%, VCAM-1 +55%, AngII +80%; all p < 0.01). Avocado supplementation mitigated these effects: BMI and feed efficiency returned to near-control levels, LDL, TG, and TC fell by 30-45%, and troponin, CK, and LDH decreased by [~]25-30%. Endothelin, VCAM-1, and AngII were reduced by 40-55% relative to L-NAME. Network analysis revealed dense pathological correlations under L-NAME (density 0.42), simplified under avocado (0.17), and most normalized with avocado+drugs (0.09), indicating restoration of physiological biomarker independence. ConclusionAvocado supplementation attenuates L-NAME-induced vascular injury by improving metabolic efficiency, correcting dyslipidemia, reducing cardiac injury, and dampening endothelial activation, while reprogramming pathological biomarker networks toward control-like organization HighlightsO_LIAvocado supplementation improves lipid balance, cardiac integrity, and vascular function in L-NAME-induced injury. C_LIO_LIContour and network analyses reveal avocado disrupts maladaptive biomarker couplings and restores control-like organization. C_LIO_LIPreclinical evidence supports avocado as a nutraceutical adjunct for integrated cardiometabolic protection. C_LI GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=152 SRC="FIGDIR/small/681762v1_ufig1.gif" ALT="Figure 1"> View larger version (36K): org.highwire.dtl.DTLVardef@12bc02borg.highwire.dtl.DTLVardef@1558740org.highwire.dtl.DTLVardef@229d49org.highwire.dtl.DTLVardef@843627_HPS_FORMAT_FIGEXP M_FIG C_FIG

Hypertension drives heterogeneous aortic remodeling, but the mechanisms underlying regional disparities remain unclear. Here, we demonstrate that adaptive immunity orchestrates spatial differences in vascular dysfunction by modulating perivascular adipose tissue (PVAT) phenotype and immune-metabolic crosstalk. Using angiotensin II (AngII)-infused wild-type (WT) and Rag1-/-mice lacking T and B cells, we integrated biaxial mechanical testing, bulk transcriptomics, and PVAT analyses. In WT mice, AngII induced pronounced descending thoracic aorta (DTA) remodeling, marked by wall thickening, reduced circumferential stiffness and inflammatory gene upregulation (Il6, Ccl2). These changes were attenuated in Rag1-/- mice, implicating T cells in thoracic maladaptation. Conversely, the infrarenal abdominal aorta (IAA) exhibited hypertensive resilience in WT mice but unmasked PPAR{gamma}-associated metabolic reprogramming (Pparg, Adipoq) in Rag1-/- mice, suggesting T cells suppress protective abdominal adaptations. PVAT heterogeneity emerged as a key regulator wherein thoracic PVAT (T-PVAT) adopted a pro-inflammatory phenotype (CCL5, TIMP-1) in WT mice, exacerbating DTA damage, while Rag1-/- mice showed thermogenic plasticity (Ucp1 upregulation) in abdominal PVAT (A-PVAT). T cell reconstitution restored maladaptive remodeling in Rag1-/- mice, confirming adaptive immunitys dual role in promoting thoracic injury and restraining metabolic resilience. This work identifies PVAT as an immune-metabolic switch governing regional susceptibility to vascular remodeling, offering spatially resolved strategies to preserve aortic compliance in hypertensive disease.

BackgroundIndividuals affected by obesity present different health trajectories and do not suffer from cardiometabolic complications all in the same way. There is a need to better understand obesity subtypes and to develop approaches for stratification. In this study we investigated both metabolomic and proteomic signatures in serum and blood plasma samples discriminating metabolically healthy from unhealthy obesity. MethodsWe investigated cross-sectional metabolomic and proteomic data from participants of the Cooperative Health Research in South Tyrol (CHRIS) study. Participants were grouped into metabolically healthy obesity (MHO) and metabolically unhealthy obesity (MUO) based on available health data in the study. A total of 461 individuals were included in the analysis, with n=130 MHO and n=331 MUO. Random forest (RF) classifiers were used to discriminate metabolically healthy from unhealthy obesity and to identify molecular features characteristic of MHO/MUO. Linear regression models were used to assess associations between each relevant metabolite/protein and MHO/MUO phenotypes independently of age, sex and body composition. ResultsThe MHO/MUO RF classifier achieved a performance of AUC = 0.709, 95% CI = (0.698,0.721). Three plasma proteins and 12 circulating metabolites were identified as relevant predictors of MHO/MUO phenotypes. Linear regression models confirmed the Apolipoprotein C-III (APOC3) association to be independent of age, visceral fat composition, medication or serum triglyceride levels. ConclusionAPOC3 was identified as a novel predictor for obesity stratification, highlighting the importance of circulating triglyceride levels in relation to metabolic health.

Hepatic glucose production is crucial for the maintenance of normal glucose homeostasis. Although hepatic insulin resistance contributes to excessive glucose production, its mechanism is not well understood. Here, we show that inositol polyphosphate multikinase (IPMK), a key enzyme in inositol polyphosphate biosynthesis, plays a role in regulating hepatic insulin signaling and gluconeogenesis both in vitro and in vivo.IPMK-deficient hepatocytes exhibit decreased insulin-induced activation of Akt-FoxO1 signaling. The expression of mRNA levels of phosphoenolpyruvate carboxykinase 1 (Pck1) and glucose 6-phosphatase (G6pc), key enzymes mediating gluconeogenesis, are increased in IPMK-deficient hepatocytes compared to wild type (WT) hepatocytes. Importantly, re-expressing IPMK restores insulin sensitivity and alleviates glucose production in IPMK-deficient hepatocytes. Moreover, hepatocyte-specific IPMK deletion exacerbates hyperglycemia and insulin sensitivity in mice fed a high-fat diet (HFD), accompanied by an increase in hepatic glucose production during pyruvate tolerance test and reduction in Akt phosphorylation in IPMK deficient liver. Our results demonstrate that IPMK mediates insulin signaling and gluconeogenesis and may be potentially targeted for treatment of diabetes. HighlightsIPMK expression is reduced in livers of HFD-fed mice. Hepatocyte-specific deletion of IPMK in mice aggravated HFD-induced insulin resistance. Loss of IPMK decreased insulin-induced activation of Akt-FoxO1 signaling, leading to the increase of glucose production in hepatocytes.

BackgroundDegenerative protein modifications (DPMs) accumulate with aging and can alter biomolecule structure and function, including via spontaneous conversion of Asn-Gly-Arg (NGR) to isoAsp-Gly-Arg (isoDGR) motifs that can bind integrins and drive chronic inflammation. Since isoDGR-modified extracellular matrix proteins are enriched in atherosclerosis and have been associated with rupture-prone plaque characteristics, we hypothesized that antibody neutralisation can inhibit key pathological features including atherosclerotic vascular plaque formation and metabolic dysfunction. MethodsWe first examined Pcmt1-/- mice which rapidly accumulate isoDGR due to lack of the corresponding repair enzyme to assess the extent of vascular protein damage. We then treated 6-8 week old atherosclerosis-prone (ApoE-/-) mice which were fed a high-fat Western diet (WD) with weekly dose of 1mg/kg isoDGR-specific monoclonal antibody (isoDGR-mAb) or isotype-matched control (while on diet) for 2 months duration. A regular chow-fed ApoE-/- group served as baseline control. Aortic atherosclerotic burden, plaque composition, systemic inflammation, lipid profiles, hepatic steatosis, and metabolic parameters (indirect calorimetry) were assessed. ResultsPcmt1-/- mice displayed extensive isoDGR deposition and degeneration of the aortic wall, linking this DPM to vascular structural damage. In the ApoE-/- mice, WD induced large aortic root plaques with abundant isoDGR and macrophage infiltration. IsoDGR-mAb treatment decreased plaque size by [~]30% with reduced lipid and collagen content (p=0.001). Furthermore, plaques in treated mice contained significantly fewer CD68+ macrophages that also exhibited limited activation. Systemically, isoDGR-mAb modified lipoprotein profiles by decreasing atherogenic VLDL/IDL/LDL cholesterol (p=0.04) while slightly increasing HDL, accompanied by a reduction in circulating inflammatory proteins. IsoDGR-mAb also protected against hepatic lipid accumulation which was reduced by [~]60% in treated animals (p<0.001), with indirect calorimetry confirming [~]30% higher oxygen consumption and energy expenditure without change in food intake or physical activity. ConclusionWe identified isoDGR as a key pathological factor involved in the progression of atherosclerosis. Remarkably, isoDGR neutralization diminished plaque inflammation and improved atherosclerotic plaque stability. Our findings support isoDGR neutralization as a promising therapeutic strategy to mitigate both atherosclerosis and aging-associated metabolic dysfunction.

Clearance of circulating insulin is critical for metabolic homeostasis. In the liver, insulin is degraded by the activity of the insulin-degrading enzyme (IDE). Here we establish a hepatic regulatory axis for IDE through WDR23-proteostasis. Wdr23KO mice have increased IDE expression, reduced circulating insulin, and defective insulin responses. Genetically engineered human cell models lacking WDR23 also increase IDE expression and display dysregulated phosphorylation of insulin signaling cascade proteins, IRS-1, AKT2, MAPK, FoxO, and mTOR, similar to cells treated with insulin, which can be mitigated by chemical inhibition of IDE. Mechanistically, the cytoprotective transcription factor NRF2, a direct target of WDR23-Cul4 proteostasis, mediates the enhanced transcriptional expression of IDE when WDR23 is ablated. Moreover, an analysis of human genetic variation in WDR23 across a large naturally aging human cohort in the US Health and Retirement Study reveals a significant association of WDR23 with altered hemoglobin A1C (HbA1c) levels in older adults, supporting the use of WDR23 as new molecular determinant of metabolic health in humans.

IntroductionInflammation is a key pathogenic feature of abdominal aortic aneurysm (AAA). Soluble epoxide hydrolase (sEH) is a pro-inflammatory enzyme that converts cytochrome P450-derived epoxides of fatty acids to the corresponding diols, and pharmacological inhibition of sEH prevented AAA formation. Both cytochrome P450 enzymes and sEH are highly expressed in the liver. Here, we investigated the role of hepatic sEH in AAA using a selective pharmacological inhibitor of sEH and hepatocyte-specific Ephx2 (which encodes sEH gene) knockout (KO) mice in two models of AAA [angiotensin II (AngII) infusion and calcium chloride (CaCl2) application]. Methods and resultssEH expression and activity were strikingly higher in mouse liver compared with aorta and further increased the context of AAA, in conjunction with elevated expression of the transcription factor Sp1 and the epigenetic regulator Jarid1b, which have been reported to positively regulate sEH expression. Pharmacological sEH inhibition, or liver-specific sEH disruption, achieved by crossing sEH floxed mice with albumin-cre mice, prevented AAA formation in both models, concomitant with reduced expression of hepatic sEH as well as complement factor 3 (C3) and serum amyloid A (SAA), liver-derived factors linked to AAA formation. Moreover, sEH antagonism markedly reduced C3 and SAA protein accumulation in the aortic wall. Co-incubation of liver ex vivo with aneurysm-prone aorta resulted in induction of sEH in the liver, concomitant with upregulation of Sp1, Jarid1b, C3 and SAA gene expression, suggesting that the aneurysm-prone aorta secretes factors that activate sEH and downstream inflammatory signaling in the liver. Using an unbiased proteomic approach, we identified a number of dysregulated proteins [e.g., plastin-2, galectin-3 (gal-3), cathepsin S] released by aneurysm-prone aorta as potential candidate mediators of hepatic sEH induction. ConclusionWe provide the first direct evidence of the livers role in orchestrating AAA via the enzyme sEH. These findings not only provide novel insight into AAA pathogenesis, but they have potentially important implications with regard to developing effective medical therapies for AAA.

BackgroundGut microbial metabolites called short-chain fatty acids (SCFA) confer protective effects against cardiovascular disease and high blood pressure. Proposed mechanisms include anti-inflammatory signalling mediated by SCFA-sensing G-protein-coupled receptors (GPCR), particularly GPR41, GPR43, and GPR109a, as suggested by knockout mouse models. We aimed to determine if rare pathogenic variants (RPVs) affecting GPCR genes in humans increase the risk of hypertension (HTN) and major adverse cardiac events (MACEs), including acute coronary syndrome, heart failure, and ischemic stroke. MethodsUsing UK Biobank whole-exome sequencing data from 393,649 European participants, we identified rare (minor allele frequency <1%) pathogenic variants with predicted high-impact functional consequences in GPCR genes, based on Ensembl Variant Effect Predictor annotations. For missense variants, pathogenicity likelihood scores from AlphaMissense, Mendelian Clinically Applicable Pathogenicity, and Combined Annotation Dependent Depletion were assessed. Multivariable logistic regression models, adjusted for age, sex, BMI, genetic ancestry, and other potential confounders, were conducted to compare RPV prevalence between cases and controls. ResultsWe identified a total of 158 RPVs in SCFA-sensing GPCR genes. The prevalence of RPV carriers was significantly higher in patients with HTN (OR=1.12, P=0.014) and MACEs (OR=1.18, P=0.009) than controls. In single GPCR gene analyses, RPVs in the FFAR2 gene (encoding GPR43) were associated with an increased risk of HTN (OR=1.23, P=0.005). RPVs in the HCAR2 gene (encoding GPR109A) were associated with a markedly increased risk of heart failure (OR=1.57, P=0.012). ConclusionsThese findings confirm and extend previous results from knockout animal models in a large population-based cohort, highlighting the potential of GPCRs as therapeutic targets for HTN and cardiovascular diseases in humans.

ObjectiveThis study aimed to investigate the epidemiological burden of metabolic dysfunction-associated steatotic liver disease (MASLD) across Cardiovascular-Kidney-Metabolic syndrome (CKM) stages and evaluate its association with cardiovascular mortality, while exploring the mediating role of insulin resistance (IR). MethodsUsing data from the National Health and Nutrition Examination Survey (NHANES, 2009-2018), we included 9,093 adults with CKM stages 1-4. MASLD was defined by validated indices (usFLI [&ge;] 30). Weighted Cox regression assessed MASLD-associated cardiovascular mortality risk. Restricted cubic splines (RCS) modeled dose-response relationships. Causal mediation analysis quantified TyG indexs contribution to MASLD-related mortality. Sensitivity analyses included subgroup stratification, missing data deleting and alternative MASLD definitions. ResultsMASLD prevalence increased significantly across advancing CKM stages (stage 1: 8.04%, stage 2:32.78%, stage 3: 41.90% and stage 4: 42.55%; P < 0.001). RCS revealed linear mortality risk escalation with rising usFLI scores (Non-line P < 0.05). MASLD independently predicted 63% higher cardiovascular mortality risk (adjusted HR=1.63, 95% CI:1.05-2.52). Stratify analyses revealed heterogeneity in associations by diabetes, CKD, CVD, and CKM stages (P for interaction < 0.05), stronger risks were observed in non-diabetic, non-CKD, non-CVD and early-stage (1-2) CKM. TyG-mediated IR explained 40.5% of MASLD-associated mortality. Sensitivity analyses confirmed robustness across MASLD definitions (FLI-based HR = 1.68, 95% CI, 1.07 - 2.63, P = 0.025). ConclusionMASLD exhibits a stage-dependent escalation in CKM populations and independently drives CVD mortality, with insulin resistance mediating 40% of this risk. Integrating MASLD screening into CKM risk stratification may enhance early intervention, particularly in early-stage patients.

ObjectiveAtherosclerosis, the main pathology underlying cardiovascular diseases is accelerated in diabetic patients. Genetic mouse models require breeding efforts which are time-consuming and costly. Our aim was to establish a new nongenetic model of inducible metabolic risk factors that mimics hyperlipidemia, hyperglycemia, or both and allows the detection of phenotypic differences dependent on the metabolic stressor(s). Methods and ResultsWild-type mice were injected with gain-of-function PCSK9D377Y (proprotein convertase subtilisin/kexin type 9) mutant adeno-associated viral particles (AAV) and streptozotocin and fed either a high-fat diet (HFD) for 12 or 20 weeks or a high-cholesterol/high-fat diet (Paigen diet, PD) for 8 weeks. To evaluate atherosclerosis, two different vascular sites (aortic sinus and the truncus of the brachiocephalic artery) were examined in the mice. Combined hyperlipidemic and hyperglycemic (HGHCi) mice fed a HFD or PD displayed characteristic features of aggravated atherosclerosis when compared to hyperlipidemia (HCi HFD or PD) mice alone. Atherosclerotic plaques of HGHCi HFD animals were larger, showed a less stable phenotype (measured by the increased necrotic core area, reduced fibrous cap thickness, and less -SMA-positive area) and had more inflammation (increased plasma IL-1{beta} level, aortic pro-inflammatory gene expression, and MOMA-2-positive cells in the BCA) after 20 weeks of HFD. Differences between the HGHCi and HCi HFD models were confirmed using RNA-seq analysis of aortic tissue, revealing that significantly more genes were dysregulated in mice with combined hyperlipidemia and hyperglycemia than in the hyperlipidemia-only group. The HGHCi-associated genes were related to pathways regulating inflammation (increased Cd68, iNos, and Tnfa expression) and extracellular matrix degradation (Adamts4 and Mmp14). When comparing HFD with PD, the PD aggravated atherosclerosis to a greater extent in mice and showed plaque formation after 8 weeks. Hyperlipidemic and hyperglycemic mice fed a PD (HGHCi PD) showed less collagen (Sirius red) and increased inflammation (CD68-positive cells) within aortic plaques than hyperlipidemic mice (HCi PD). HGHCi-PD mice represent a directly inducible hyperglycemic atherosclerosis model compared with HFD-fed mice, in which atherosclerosis is severe by 8 weeks. ConclusionWe established a nongenetically inducible mouse model allowing comparative analyses of atherosclerosis in HCi and HGHCi conditions and its modification by diet, allowing analyses of multiple metabolic hits in mice.

BackgroundWhile excess salt intake is known to affect cardiovascular health, its role in nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH) is less established. We aimed to examine the longitudinal association between salt intake and incident NAFLD/NASH. Methods and findingsThis study included 494,170 UK Biobank participants without NAFLD/NASH at baseline. Salt exposure was assessed via self-reported salt-adding frequency (four-point Likert scale) and estimated 24-hour sodium intake from spot urine. Incident NAFLD/NASH cases were defined by diagnostic codes. Hazard ratios (HRs) and 95%CIs were calculated by using Cox proportional hazards models. Mendelian randomization and mediation analyses were conducted to infer causality and explore underlying mechanisms. Over a mean 14.3-year follow-up, 7,307 NAFLD and 630 NASH cases were identified. Both higher salt-adding frequency and sodium intake were significantly associated with elevated NAFLD/NASH risk. Compared to those who never/rarely added salt, adjusted HRs (aHRs) for NAFLD were 1.02 (95% CI: 0.98-1.10), 1.20 (1.09-1.31), and 1.31 (1.16-1.48) for sometimes, usually, and always, respectively (P-trend < 0.001). Always adding salt was also linked to higher NASH risk (aHR = 1.42; 95% CI: 1.01-1.99). Per 1g increase in estimated 24-hour sodium intake, NAFLD and NASH risks increased (aHRs = 1.82; 95% CI: 1.73-1.92 and 2.17; 95% CI: 1.81-2.61). Genetically predicted salt-adding frequency was also associated with increased NAFLD risk (Odds ratio = 1.54; 95% CI: 1.16-2.05). The salt-NAFLD associations were more pronounced among individuals with normal body-mass index (BMI) or normal alanine transaminase (P-interaction = 0.002, 0.005), with BMI mediating 16.4% (95% CI: 13.1-22.4%) of the salt-NAFLD association. ConclusionsHigher salt intake is independently associated with increased NAFLD/NASH risk, particularly in metabolically healthy individuals. These findings support liver-focused strategies in salt reduction policies.

Background and aimsThis study investigated the prevalence and causal relationships of chronic metabolic diseases (diabetes, hypertension, and dyslipidemia) with steatotic liver disease (SLD), specifically metabolically associated alcoholic liver disease (MetALD). MethodsWe conducted a comprehensive analysis using cross-sectional data from the Korea National Health and Nutrition Examination Survey (KNHANES) from 2011 to 2021 and the National Health and Nutrition Examination Survey (NHANES) from 1999 to 2020. Longitudinal data from 2001 to 2014 from the Korean Genome and Epidemiology Study (KoGES) were used. Participants were categorized into the metabolic dysfunction-associated SLD (MASLD), MetALD, and ALD groups based on their hepatic steatosis index (HSI) and alcohol consumption. Logistic and Cox regression analyses were performed to assess the prevalence and incidence of chronic diseases. ResultsIn both the KNHANES and NHANES cohorts, an increased HSI was significantly associated with a higher prevalence of chronic metabolic diseases. Longitudinal data from the KoGES cohort showed that MASLD and MetALD were significant predictors of chronic metabolic disease in both men and women. MetALD showed a higher hazard ratio for the development of chronic metabolic diseases than MASLD in Cox regression analysis. ConclusionThis study highlighted the intertwined nature of SLD and metabolic health, with an emphasis on the role of MetALD. The significant association between MetALD and chronic metabolic diseases underscores the need for integrated management strategies that address both liver health and metabolic risk factors.