Using a large elderly population cohort, we investigated the relationship between choroid plexus (CP) volume, a potential indicator of glymphatic system activity, and lifestyle-related vascular risk and protective factors, as well as age-related changes in CP volume. This cross-sectional study included 2137 participants (median age, 70 years) who underwent 3-T MRI and vascular risk factor assessment. Factors included body mass index (BMI), systolic blood pressure (SBP), total cholesterol, hemoglobin A1c (HbA1c), smoking (Brinkman Index), and physical activity (International Physical Activity Questionnaire). Multiple regression analyses were conducted, adjusting for total intracranial volume (TIV), age, gender, and medication use. CP volume significantly increased with age (r = 0.351, p < 0.001). Among the vascular risk factors, only a higher Brinkman Index (smoking) was significantly associated with a larger CP volume in the overall analysis (β = 0.075, FDR-corrected p < 0.001). In sex-specific analyses, higher systolic blood pressure and an increased Brinkman Index were associated with larger CP volumes in males. In females, no vascular risk factors remained significant after correction for multiple comparisons. These factors explained only a small proportion of the variance in CP volume (R² = 0.013). In this large cohort of older adults, choroid plexus volume increases with age. Sex-specific analyses revealed that systolic blood pressure and smoking were significant risk factors in males, while no significant associations were found in females. However, these vascular factors explained only a small proportion of the variance in CP volume. Question How do lifestyle-related vascular risk factors like hypertension, smoking, and obesity affect choroid plexus volume, a potential indicator of glymphatic function? Findings In this elderly cohort, choroid plexus volume increased with age and was associated with smoking; males showed an additional significant association with higher blood pressure. Clinical relevance Understanding the sex-specific links between vascular risk factors and choroid plexus volume is crucial for developing targeted strategies to preserve glymphatic function and brain health in aging populations.

Elevated childhood blood pressure (BP) is a known predictor of future adult cardiovascular risk. We explored whether baseline BP predicted vascular health decline over time. We included 715 children with baseline (mean age, 7 years) and 4-year follow-up data. Measures of central hemodynamics included pulse wave velocity, cardiac output, cardiac index, stroke volume, augmentation index corrected for heart rate of 75 bpm, and total vascular resistance. Kaplan-Meier survival analyses were used to compare the event rate of follow-up vascular measures greater than or equal to the highest population quartile for each measure between BP groups at baseline. In the total population, we determined the relative risk of baseline BP predicting poorer vascular health at follow-up using Cox proportional hazard models. At baseline, pulse wave velocity, stroke volume, cardiac output, and augmentation index corrected for heart rate of 75 bpm were higher in children in the elevated BP compared with the normotensive group (all P≤0.014). In the total population, baseline systolic BP increased the relative risk for higher pulse wave velocity (HR=1.98; P<0.001), cardiac output (HR=1.23; P=0.005), and stroke volume (HR=1.21; P=0.008) at follow-up, while diastolic BP increased the relative risk for higher follow-up pulse wave velocity (HR=1.57; P=0.038), augmentation index corrected for heart rate of 75 bpm (HR=1.24; P=0.003), and total vascular resistance (HR=1.17; P=0.019). Elevated BP in prepubescent children may contribute to early functional changes in central hemodynamics preceding hypertension in adolescence. These findings suggest a high-volume load in at-risk children, which may contribute to the onset of early vascular aging and hypertension in early life.

Relevance of Early Vascular Aging in Clinical Routine.

High estimated pulse-wave velocity is associated with lower brain white matter microstructural integrity twelve years later.

BackgroundIndirect estimates of pulse wave velocity (PWV) have been proposed as a feasible alternative for PWV assessment in clinical practice; however, their validity and clinical applicability remain uncertain. This study aimed to evaluate the relationships between indirect measures of arterial stiffness and directly measured PWV to determine their potential utility in clinical settings.MethodsIn this multicentre, international study, data from 4,206 individuals from Brazil, Greece, Korea, and Australia were analysed. The relationships between estimated PWV (ePWV), 24-hour (24h)-pulse pressure (PP), Early Vascular Aging Ambulatory Score (EVAAS), and carotid-femoral (cf-PWV) and/or brachial-ankle (ba-PWV) PWV were assessed through correlation and multivariate linear regression analyses. Subgroup-specific associations were also examined.ResultsThe study population had a mean age of 57.6 ± 14.3 years, with 42.5% being male and 82.1% having pre-existing hypertension. After adjusting for multiple factors related to arterial stiffness, ePWV demonstrated a strong association with cf-PWV (β = 0.599, P < 0.001) and ba-PWV (β = 1.342, P < 0.001). 24h-PP and EVAAS showed moderate associations with both cf-PWV and ba-PWV. Subgroup analyses indicated that ePWV correlated more strongly with both cf-PWV and ba-PWV in individuals without traditional cardiovascular risk factors.ConclusionsePWV may be used as a surrogate marker for arterial stiffness, particularly in individuals without major cardiometabolic comorbidities. Although 24h-PP and EVAAS are also associated with PWV, their clinical utility varies across subgroups. Future research should explore their role in improving cardiovascular risk prediction and guiding personalized treatment strategies for vascular aging.Trial RegistrationPROSPERO Identifier: CRD420250618863

Background The concept of Cardiovascular-Kidney-Metabolic syndrome (CKM), an intricate interplay among metabolic risk factors, chronic kidney disease (CKD), and the cardiovascular system, was introduced by the American Heart Association (AHA). This indicates a markedly increased susceptibility to various organ dysfunctions and adverse cardiovascular events. In CKM management, reliable markers are critical. The purpose of this study was to investigate the relationship between ePWV and mortality and prognosis in patients with CKM. Methods This study included 14,372 participants from the National Health and Nutrition Examination Survey (NHANES) from 2005 to 2018. ePWV assessed vascular aging and arterial stiffness. The association between ePWV and CKM, as well as their risk of death, was analyzed using weighted logistic regression and Cox proportional hazards models. Bootstrap sampling was used to analyze the mediating effect of the Triglyceride-Glucose Index (TyG) between ePWV and CKM. The Stage 4 ePWV threshold was further validated using an independent retrospective cohort of 3,157 patients from the cardiology department of a tertiary hospital in southern China. Results The RCS analysis demonstrated a nonlinear positive association between estimated pulse wave velocity (ePWV) and cardiovascular–kidney–metabolic syndrome (CKM) (P &amp;lt; 0.01). Subgroup analysis indicated that this association was strongest among young adults aged 20–40 years (P &amp;lt; 0.01), whereas no significant association was observed in elderly individuals over 75 years old (P = 0.791). Kaplan–Meier curves showed that individuals with higher ePWV had significantly worse survival than those with lower ePWV. Further mediation analysis revealed that the triglyceride–glucose (TyG) index mediated 25.23% of the association between ePWV and CKM. In the external validation cohort, the ePWV threshold corresponding to CKM Stage 4 was 8.52, which was consistent in direction and statistical significance with the NHANES-derived model. Conclusion Elevated ePWV is nonlinearly associated with increased CKM risk, particularly among young adults. This finding provides insights into managing arterial stiffness and preventing CKM progression in individuals at high risk of early cardiovascular disease.

To observe the effect of mild moxibustion on microRNA-34a (miR-34a)/silent information regulator 1 (sirt1)/tumor protein p53 (p53) signaling pathway in the thoracic aorta and kidney of senescent rats, so as to investigate the mechanism of mild moxibustion in attenuating vascular and renal aging. Fifty 8-month-old male SD rats were given intraperitoneal injection of D-galactose (300 mg·kg-1·d-1) for 4 weeks to establish aging model. Twenty-four successfully modeled rats were randomly divided into aged control, medication and mild moxibustion groups (with 8 rats in each group), with 8 additional 3-month-old male SD rats as the young control group. Rats of the mild moxibustion group received mild moxibustion at "Guanyuan" (CV4) and bilateral "Shenshu" (BL23) once daily for 5 d/week over 8 weeks. Rats of the medication group were intraperitoneally injected with testosterone propionate (7 mg/kg) every 3 d for 8 weeks. The aged and young control groups received equivalent volumes of 0.9% normal saline. Exhaustive swimming time was measured pre- and post-treatment. Histomorphology and collagen content of the thoracic aorta and kidney were assessed using HE and Masson staining. Serum total testosterone (TT) and free testosterone (FT) contents were detected by ELISA. Klotho protein expression in aortic and renal tissue was detected via immunohistochemistry. The relative protein expressions of sirt1, p53, and transforming growth factor-β1 (TGF-β1) were detected using Western blot, while real-time PCR was used to detect miR-34a, sirt1, and p53 mRNA expressions. Compared with the young control group, the exhaustive swimming time of rats in the aged control group shortened (P<0.01), collagen deposition in aortic and renal tissues increased (P<0.01), serum TT/FT content reduced (P<0.01), the positive expression of Klotho, the protein and mRNA expression levels of sirt1 in aortic and renal tissue decreased (P<0.01), and the expression levels of p53 protein and mRNA, TGF-β1 protein, and miR-34a mRNA increased (P<0.01). Mild moxibustion and medication interventions reversed these trends:both groups showed prolonged swimming time (P<0.01), reduced collagen (P<0.01), elevated serum TT/FT content (P<0.01), and down-regulated p53 protein, miR-34a mRNA in aortic and renal tissue (P<0.01), TGF-β1 protein and p53 mRNA in aortic tissue (P<0.05, P<0.01), and mRNA expression level of sirt1 in renal tissue (P<0.01). Compared with the medication group, the collagen deposition of aortic tissue in the mild moxibustion group decreased (P<0.05), positive expression of Klotho in renal tissue, and protein expression level of sirt1 in aortic tissue increased (P<0.05), the protein expression level of p53 in renal tissue decreased (P<0.05), while the mRNA expression levels of p53 in aortic tissue and miR-34a in renal tissue decreased (P<0.05). HE staining showed thickened thoracic aorta and reduced glomeruli number in the aged control group, which was improved in the medication and mild moxibustion groups, including thinner thoracic aortic intima, more complete glomerular structure, and more regular renal icles and glomerular lumen, and the improvement in the mild moxibustion group was better than that in the medication group. Mild moxibustion exerts anti-aging effects on the thoracic aorta and kidney of senescent rats, which may be related with its effect on modulating the miR-34a/sirt1/p53 signaling pathway.

BackgroundEarly vascular aging (EVA) is a key contributor to the elevated risk of cardiovascular disease, which remains a leading cause of mortality and disability in patients with type 2 diabetes mellitus (T2DM). This study aims to investigate the association between the triglyceride-glucose (TyG) index and EVA in T2DM patients.MethodsT2DM patients were enrolled, and their clinical data were collected. Bilateral brachial-ankle pulse wave velocity (baPWV) was measured and used to determine the presence of EVA. Binary logistic regression analysis was conducted to evaluate the effect of the TyG index and identify factors associated with EVA. A predictive model was constructed using a nomogram, and its performance was assessed using receiver operating characteristic (ROC) curve analysis.ResultsA total of 380 patients were included in the study, among whom 144 (37.89%) were identified as having EVA. After adjusting for all relevant covariates, the TyG index remained independently and positively associated with EVA (odds ratio [OR] = 2.76, 95% confidence interval [CI]: 1.57–4.85), patients in the highest TyG tertile had a 4.30-fold increased risk of developing EVA compared to those in the lowest tertile (95% CI: 1.97–9.42). Elevated TyG index (OR = 2.53, 95% CI: 1.79–3.57) and systolic blood pressure (OR = 1.04, 95% CI: 1.02–1.06) were positively associated with EVA in T2DM patients, while male sex (OR = 0.50, 95% CI: 0.30–0.83) and absence of T2DM complications (OR = 0.56, 95% CI: 0.34–0.92) were negatively associated with EVA. Based on these findings, a predictive model was developed, which demonstrated good discrimination with an area under the ROC curve (AUC) of 0.776 (95% CI: 0.728–0.824).ConclusionsA higher TyG index and elevated systolic blood pressure were independently associated with the presence of EVA, whereas male sex and the absence of T2DM complications were associated with a lower likelihood of its occurrence. These findings provide valuable insights for the early identification of high-risk individuals.

Terazosin (TZ), a well-known antagonist of the α1-adrenergic receptor (α1-AR), has demonstrated protective effects on vascular endothelial cells (ECs) and reduced vascular stiffness in clinical studies. Endothelial dysfunction and oxidative stress are central drivers of cardiometabolic diseases such as diabetes, where sustained ROS burden accelerates EC senescence and barrier failure. These findings suggest its potential role in combating vascular aging and atherosclerosis; however, the underlying mechanisms remain partially understood. In this study, we investigated whether TZ can prevent atherosclerosis in ApoE-/- mice fed a high-cholesterol diet and aimed to elucidate the mechanisms involved. Our results showed that TZ significantly reduced plaque size, EC senescence, vascular permeability, and reactive oxygen species (ROS) levels, effectively inhibiting atherosclerosis independently of α1-AR signaling. In cultured primary human umbilical vein ECs (HUVECs), TZ inhibited EC senescence via the Pgk1/Hsp90 pathway. It enhanced the interaction between Hsp90 and the antioxidant enzyme peroxiredoxin 1 (Prdx1), leading to lower ROS levels-a key driver of cellular senescence. These findings were confirmed in atherosclerotic ApoE-/- mice. Furthermore, senescent ECs exhibited increased levels of vascular endothelial growth factor A (VEGFA) and decreased levels of angiostatin, contributing to higher vascular permeability and exacerbating atherosclerosis. TZ effectively reversed these changes. Overall, our study demonstrates that TZ primarily alleviates EC senescence and atherosclerosis through the Pgk1/Hsp90/Prdx1 pathway, highlighting Pgk1 activation as a strategy that may also mitigate endothelial dysfunction and oxidative stress in broader cardiometabolic contexts (e.g., diabetes), suggesting that TZ is a promising senomorphic agent for treating vascular aging and atherosclerosis in clinical settings and that Pgk1-targeted interventions could have implications beyond atherosclerosis.

Vascular ageing (VA), characterised by vascular endothelial dysfunction, is a major contributor to age-related chronic conditions. Leucine-rich repeat-containing protein 8A (LRRC8A) is vital in maintaining vascular endothelial function; however, the role of endothelial LRRC8A in VA is undefined. We aimed to investigate the role and mechanism of endothelial LRRC8A in VA. We found that LRRC8A expression was clearly downregulated in the aged murine aortas. Further integrated analysis of single-cell and bulk RNA-seq and experimental verification revealed that endothelial LRRC8A governed VA by counteracting cell cycle, cellular senescence and oxidative stress. Additionally, endothelial LRRC8A deletion exacerbated the D-galactose (D-gal)-induced VA progression. Mechanistically, endothelial LRRC8A phosphorylated AMPK at T172 and subsequently facilitated SIRT1 nuclear translocation, ultimately counteracting the p53-dependent senescence pathway and activating the FOXO3-dependent antioxidant pathway. Therapeutically, pharmacological agonists of AMPK and SIRT1 effectively rescued endothelial cell senescence and VA in the context of endothelial LRRC8A deficiency. Additionally, endothelial-targeted adeno-associated virus (AAV)-LRRC8A gene therapy can effectively delay the progression of VA in naturally ageing mice. Our findings provide the first evidence supporting endothelial LRRC8A as a novel modulator of the AMPK-SIRT1 axis and suggest that targeting LRRC8A represents a promising therapeutic strategy for VA and age-related chronic conditions.

Despite effective viral suppression with antiretroviral therapy (ART), people living with HIV (PLWH) experience persistent inflammation, immune dysfunction, and premature onset of cardiovascular and aging-related comorbidities. To define the underlying mechanisms, we performed longitudinal transcriptomic profiling in peripheral blood mononuclear cells (PBMCs) from a cohort of simian immunodeficiency virus (SIV)-infected rhesus macaques spanning four key stages: pre-infection, acute infection, short-term ART, and long-term ART. Bulk RNA sequencing revealed dynamic immune remodeling across infection and treatment. Acute SIV infection induced robust antiviral and inflammatory programs, with upregulation of interferon-stimulated genes (ISGs), IL-27, JAK/STAT, and NF-κB signaling, coupled with suppression of T- and B-cell activation pathways. Short-term ART effectively reversed these transcriptional perturbations, restoring adaptive immune gene expression and reducing innate antiviral responses to near-baseline levels. In contrast, chronic SIV infection on long-term ART maintained viral suppression but was characterized by reactivation of innate immune pathways, including TLR2/TLR4/MYD88, NF-κB, and inflammasome (NLRP3/or NLRP12, caspase-1) signaling, along with sustained macrophage activation, platelet/coagulation signaling, and senescence-associated secretory phenotype. Protein analyses confirmed persistent CASPASE-1 and NF-κB activation in spleen tissue. Pathologic evaluation of a carotid artery from an SIV-infected, long-term ART–treated macaque revealed macrophage-rich plaques with p21+ senescent cells with intraluminal thrombus formation, recapitulating key features of HIV-associated atherogenesis. Together, these findings demonstrate that while ART normalizes acute infection–induced immune dysregulation, chronic SIV infection sustains a chronic, macrophage- and TLR-driven inflammatory state linked to vascular injury and aging process regardless of long-term suppression of viremia. Targeting inflammasome, NF-κB, and senescence pathways may mitigate non-AIDS comorbidities in PLWH.

Vascular endothelial cell senescence is closely associated with cardiovascular disease. The deficiency of natriuretic peptide receptor A (NPRA), encoded by the natriuretic peptide receptor 1 (NPR1) gene, contributes to vascular endothelial aging, but the cause of its reduced expression is unclear. In this study, we performed reverse chromatin immunoprecipitation (R-ChIP) to analyze the binding proteins of the NPR1 promoter and found that nucleolar and coiled body phosphoprotein 1 (NOLC1) functions as a key regulator of NPRA transcription in endothelial cells. Similar to NPRA, NOLC1 is also expressed at reduced levels in senescent endothelial cells. Knockdown of NOLC1 decreased both NPRA mRNA and protein expression levels. Furthermore, inhibition of NOLC1 expression triggered cellular senescence hallmarks, including elevated p53/p21 levels, enhanced SA-β-gal activity, ROS accumulation, G0/G1 cell cycle arrest, and impaired migration. NPRA overexpression rescued senescence and dysfunction in NOLC1-deficient cells, restoring proliferative and migratory capacity. In conclusion, our findings demonstrate that nucleolar phosphoprotein NOLC1 is a key regulator of NPRA transcription in endothelial senescence.

Aging is a complex, multifaceted process affecting all organ systems, with vascular aging playing a central role in organismal health decline. Beyond its role in circulation, the vascular system acts as a dynamic interface between tissues, influencing countless physiological functions such as tissue regeneration and repair, immune responses, and metabolic balance. Importantly, age-related vascular impairment-characterized by a peculiar set of endothelial aging hallmarks-exacerbates age-related diseases (ARDs) such as cardiovascular disorders, neurodegeneration, chronic kidney disease, sarcopenia, and osteoporosis. This review combines basic concepts of angioscience and aging biology with translational interventions to devise clinical strategies promoting a functional rejuvenation of old and compromised blood vessels, fostering the prevention, delay or treatment of ARDs. Starting from the description of the cellular and molecular mechanisms driving vascular aging, a cutting-edge perspective on the organ-specific vascular impairment and its impact on tissue function is offered. Given the central role of the vasculature in aging, how targeting vascular aging through pharmacological, genetic, and lifestyle interventions holds promise for mitigating its systemic consequences and improving healthspan is discussed. Finally, how the combination of animal models (e.g., parabiosis) and novel microphysiological systems, coupled with multi-omics and artificial intelligence-driven analyses, is advancing the field toward the identification of strategies that promote vascular resilience and extend healthspan, addressing one of the most pressing biomedical challenges of a worldwide aging population is highlighted.

Introduction: Vascular aging (VA) refers to the progressive deterioration of vascular structure and function with age. Vascular age exceeding chronological age indicates higher cardiovascular risk. Indices such as brachial-ankle pulse wave velocity (baPWV), carotid intima-media thickness (cIMT), carotid plaque, and ankle-brachial index (ABI) are often used to assess VA. Research questions: The comparative performance of VA models constructed using these different indicators has not yet been evaluated. This study aimed to compare the cardiovascular risk predictive value of VA models based on baPWV, cIMT, carotid plaque, and ABI. Methods: This study included participants from a Chinese community-based prospective cohort, excluding those with baseline stroke or myocardial infarction. Vascular age was calculated from multivariable regression models incorporating traditional cardiovascular risk factors, medications, and a vascular index (baPWV, cIMT, carotid plaque, or ABI). Residuals between vascular age and chronological age were defined as ΔAge. VA phenotypes were classified as supernormal VA, normal VA (NVA), or early VA (EVA) based on the 10th and 90th percentiles of ΔAge. The endpoint was major adverse cardiovascular events (MACE), including cardiovascular mortality, nonfatal stroke, and nonfatal myocardial infarction. Restricted cubic spline analyses and Cox proportional hazards models were used to examine the association of ΔAge and VA phenotypes with MACE. Results: A total of 3,561 participants (mean chronological age: 58.7±8.5 years) were included. During a mean follow-up of 7.2±1.3 years, 294 (8.26%) participants experienced MACE. The VA model based on baPWV showed the highest explanatory power (R 2 = 0.505) (Table 1).ΔAge based on baPWV showed a backwards L-shape association with MACE (Figure 1A). After adjustment, each standard deviation increase in ΔAge based on baPWV was associated with a 14% higher risk of MACE (HR 1.14, 95% CI 1.01–1.27, P=0.027; Table 2). EVA group based on baPWV had a significantly higher risk of MACE compared with NVA group (HR 1.44, 95% CI 1.05–1.97, P=0.023; Table 3). In contrast, VA phenotypes by other indices were not significantly associated with MACE (Table 3). Conclusions: Vascular age based on baPWV demonstrated superior cardiovascular risk prediction versus models using cIMT, plaque, or ABI. baPWV may serve as the preferred method for vascular aging assessment in cardiovascular risk stratification.

Introduction: Healthy Vascular Aging (HVA) refers to the preservation of vascular function with advancing age, challenging the common belief that hypertension and increased arterial stiffness are unavoidable outcomes of aging (Hypertension, PMID: 28559398). This definition identifies individuals who retain young vascular characteristics beyond midlife. While traditional clinical metrics are used to assess HVA, waveform-derived biomarkers, such as cardiovascular intrinsic frequencies (IFs), may provide a non-invasive and physiologically grounded alternative. IFs extracted from carotid pressure waveforms have previously been shown to be associated with cardiovascular function and vascular health (Hypertension, PMID: 33390053). In this study, we examine the association between carotid-derived IFs and the HVA status. Methods: The study sample was drawn from the Original (Exam 26), Offspring (Exam 7), and Third Generation (Exam 1) cohorts of the Framingham Heart Study. As established in the literature, we assigned HVA to the individuals with age ≥ 50 years, systolic blood pressure &lt;140 mmHg, diastolic blood pressure &lt;90 mmHg (without hypertension medication), and a carotid-femoral pulse wave velocity &lt;7.6 m/s. After applying these criteria, a total of N = 2,242 participants remained for analysis. Carotid pressure waveforms were acquired using applanation tonometry. We used these waveforms and computed the IF metrics for each individual: ω 1c (IF during systole, related to left ventricular inotropic function and LV-arterial coupling) and ω 2c (IF during diastole, related to arterial function). The associations between the HVA status and carotid-derived IFs were assessed using box-and-whisker plots and the Mann–Whitney U test. Results: The carotid-based IF metrics showed statistically significant differences between individuals with and without HVA (p &lt; 0.0001 for all comparisons). The group-wise distributions of these parameters are illustrated in Figure 1, highlighting distinct shifts in IF values between the HVA and no-HVA groups. Lower ω 1c but higher ω 2c were observed in the HVA group. Conclusion: Our results demonstrated that Intrinsic frequencies of carotid pressure waveforms are associated with the status of vascular aging. The findings of this study support the potential utility of IFs as a non-invasive approach to assess vascular health aging.

Background: Smooth muscle cells (SMC), an important cellular component of atherosclerosis, exhibit variant phenotypes after transdifferentiation. Their differentiation into fibromyocytes (FM) is regulated by the TCF21 transcription factor. The TCF21 gene was associated with decreased cardiovascular risk in a genome wide association study. However, RNA seq data suggesting a pro-inflammatory role of TCF21. Cell senescence is an irreversible cell proliferation arrest associated with vascular aging, and the progression of atherosclerosis. Hypothesis: We hypothesized that FM promote cellular senescence and exacerbate formation of the plaque’s necrotic core contributing to atherosclerotic plaque progression. Methods: We analyzed single-cell RNA seq (scRNAseq) datasets obtained for human coronary arteries, APOE-null mouse aortas, and a spatial transcriptomics (ST) dataset obtained for pig coronary artery. A senescence module score was calculated based on expression levels of 150+ senescence genes using a gene set enrichment algorithm. Spearman coefficients were calculated to align each gene expression pattern with senescence score distribution. Results: Gene expression profiling of atherosclerotic specimens obtained from humans, pigs and mice revealed that FM exhibited higher senescence module scores suggesting increased FM susceptibility to developing senescence. Cell type ratios were estimated for ST spots by deconvolution. FM was the most abundant cell type in the porcine coronary plaque’s fibrous cap (FC). Complement factor H (CFH) and versican (VCAN) were identified as top genes expressed in FM and highly correlated with senescence module scores: expression level of CFH and VCAN were 13 and 7.5-fold higher in FM compared to SMC, respectively. VCAN is highly expressed in senescent cells potentially promoting senescence of neighboring cells. CFH inhibits immune cell-mediated efferocytosis to clear dying cells, which may exacerbate formation of the necrotic core. Conclusions: Our results identified high levels of FM in FC in advanced coronary plaque and indicate that FM are prone to develop senescence. Upregulation of VCAN and CFH in FM are potential mechanisms promoting senescence and extension of the necrotic core. These findings revealed a novel role of FM in plaque progression and destabilization and identified FM as novel cellular target for development of anti-atherosclerotic therapy.

Background: Pulse pressure (PP), the difference between systolic and diastolic blood pressure, reflects arterial stiffness and vascular aging. Its role in predicting gastrointestinal (GI) bleeding risk among anticoagulated atrial fibrillation (AF) patients remains poorly defined. Objective: To assess whether elevated pulse pressure independently predicts GI bleeding in anticoagulated patients with atrial fibrillation. Methods: This retrospective cohort study used the REACHnet database. Adult patients diagnosed with AF and receiving blood thinners were included. PP was derived from outpatient systolic and diastolic measurements and categorized into tertiles: T1 (&lt;46 mmHg), T2 (46–62 mmHg), and T3 (&gt;62 mmHg). The primary outcome was GI bleeding requiring hospitalization. Demographic and clinical variables were compared across tertiles using Wilcoxon tests, t-tests, and Chi-square tests as needed. Kaplan-Meier analysis assessed time to GI bleeding. Cox proportional hazards models were adjusted for confounders, including systolic blood pressure (SBP). Results: Among 3,142 patients (mean age 74 ± 8.9 years; 47% female), 9.9% experienced GI bleeding over a median follow-up of 4.1 years. Patients in the highest PP tertile (T3) had significantly greater prevalence of hypertension, diabetes, peripheral artery disease, history of stroke, and anticoagulant use (all p&lt;0.01) (Table 1). Despite this higher burden of comorbidity, elevated PP remained an independent predictor of GI bleeding. Adjusted analysis showed that patients in T3 had a 48% increased risk of GI bleeding compared to T1 (HR: 1.48; 95% CI: 1.19–1.84; p&lt;0.001), independent of SBP and other clinical factors. Kaplan-Meier curves demonstrated significantly lower bleeding-free survival in T3 (log-rank p=0.003) (Figure 1). Conclusion: High pulse pressure, especially &gt;62 mmHg, is independently associated with GI bleeding in anticoagulated AF patients. This association persists after adjustment for SBP and other comorbidities. PP should be considered in bleeding risk assessment models in clinical practice.

Background: The hallmark of high-risk atheroma is presence of a large lipid/necrotic core (LNC) containing macrophages (MF), foam cells and necrotic tissue. Enlargement of the core contributes to plaque rupture. Cell senescence is an irreversible cell proliferation arrest associated with vascular aging. Senescent MF are reported to be present in atherosclerotic plaques; however, their functional role remains to be elucidated. Hypothesis: Senescent MF promote expansion of the LNC contributing to plaque destabilization. Methods: Coronary sections were obtained from atherosclerotic hypercholesteremic pigs. LNCs were identified by staining of cryosections with Oil Red. MF cell marker (scavenger receptor A, SRA), and senescence markers (β-galactosidase (βGal) and γ-H2AX histone) were quantified by immunohistochemistry. βGal activity was assessed with X-Gal assay. Spatial transcriptomics (ST) and single cell RNA seq (scRNA seq) analysis was used to identify differentially expressed genes (DEGs) and quantify senescence module scores in plaque compartments. Results: Increased βGal activity (2.5-fold increase) and upregulated γ-H2AX histone levels (1.8-fold increase) (both are P&lt;0.05) were detected in cells inside LNCs and in a 50um-thick plaque area surrounding LNCs. The number of senescent MF (SRA+/βGal+ cells) was increased (P&lt;0.05) in the area surrounding LNCs compared to other plaque compartments. Senescent marker positivity was higher in cells in areas surrounding LNCs in plaques with large cores (&gt;40% of core area normalized per plaque area) compared to plaques with smaller cores (&lt;15%). These results suggest that development of senescence correlated with LNC size. Top DEGs in cells inside LNCs were molecules associated with induction or development of cell senescence (cathepsin D, cystatin 3 and ferritin light chain) and MF/foam cell markers (SRA, CD68 and lipoprotein lipase). Senescence module score was higher in LNCs vs. other compartments. scRNA seq and ST data were used to identify and locate MF subtype contributing to senescence in LNCs. M2 polarized MF have higher senescence module scores compared to M1 MF. M2 MF were highly enriched in the area between the LNC and plaque fibrous cap. Conclusions: Our data suggest that upregulated levels of senescent MF in LNCs and in the surrounding area promote expansion of the LNC and contribute to plaque destabilization. These results identify senescent MF as a potential novel therapeutic target.

Vascular aging is marked by increased levels of reactive oxygen species in endothelial cells. Reactive oxygen species can, among others, be produced by dysfunctional mitochondria, contributing to acceleration of vascular aging by promoting DNA damage response and senescence. In the aged vasculature, impaired endothelial cell function causes decreased vasodilation, which may also have an impact on peripheral organs such as the kidney. The aim of this study was to investigate the effect of chronic treatment with SUL-138 (30 mg/kg/day), a novel mitochondrial protective compound, on DNA damage-prompted, accelerated endothelial aging and associated kidney dysfunction in mice. Endothelial-specific aging was induced by knock-out (KO) of DNA repair endonuclease Ercc1 in mice [endothelial-cell specific Ercc1 KO (EC-KO) mice]. We showed that impaired endothelium-dependent vasodilation and expression of DNA damage response markers in EC-KO mice were restored after the treatment with SUL-138. The underlying mechanism of improved vasodilation was an increase in endothelium-derived hyperpolarization (EDH). Endothelial-specific aging induced tubular injury, sodium wasting, and increased inflammatory markers in the kidney, which were normalized by the treatment with SUL-138. We conclude that accelerated endothelial aging adversely affects vascular function and causes kidney tubular injury. SUL-138 rescues endothelial aging, restores vasodilation by increasing EDH, and protects the kidney. Thus, preservation of mitochondrial function is a potential pharmacotherapeutic target in aging-related dysfunction provoked by the DNA damage response.

Vascular aging is characterized by a tandem increase in pulse pressure and large elastic artery stiffness. Greater stiffness of the large arteries leads to elevated pulse pressure transmitted into the cerebral circulation, causing dysfunction. However, little is known in females about age-related stiffening of the arteries and the impacts of high pulse pressure on the cerebral vasculature. To examine the effects of sex and age on the cerebral artery response to pulse pressure, we studied cerebral arteries collected from young and old female and male C57BL/6 mice. Isolated posterior cerebral arteries (141.3±3.9 μm diameter), were exposed ex vivo to acute static pressure, low pulse pressure, and high pulse pressure. Exposure to high pulse pressure impaired endothelium-dependent dilation in cerebral arteries from young female and male mice, with impairments also occurring in young female cerebral arteries after exposure to low pulse pressure. In contrast, exposure to low or high pulse pressure had minimal effects on cerebral artery endothelium-dependent dilation in old male and female mice. During exposure to high pulse pressure, young females had higher cerebral artery compliance compared with young males and old females. Old mice had higher cerebral artery passive stiffness and aortic pulse wave velocity compared with young mice. We also found age and sex differences in arterial wall thickness, collagen and elastin content, and matrix metalloproteinase 9 expression. Taken together, young female mice have more compliant cerebral arteries, which are more susceptible to endothelial dysfunction caused by pulse pressure.