Targeting biological processes of aging is a central goal of geroscience; however, limited data exist regarding the feasibility of incorporating biological aging biomarkers into dietary intervention trials. We conducted a pilot feasibility study among 34 adults aged 48-81 years with metabolic syndrome, a condition associated with elevated risk of age-related cardiometabolic disease and advanced biological aging. Participants consumed 1 oz of tree nuts and two tablespoons of extra virgin olive oil daily for 4 weeks. The primary objectives were to evaluate feasibility, adherence, and participant acceptability of epigenetic aging assessments. Exploratory outcomes included DunedinPACE, a measure of the pace of aging, and AgeAccelGrim, a measure of biological age relative to chronological age. At baseline, all participants exhibited a faster pace of biological aging than average as assessed by DunedinPACE, supporting metabolic syndrome as a promising target population for geroscience interventions. Adherence to the dietary intervention exceeded 95%, and most participants reported willingness to participate in a similar longer-term trial. Participants expressed a strong interest in learning their biological age and indicated that evidence of slowed aging would motivate sustained dietary change. No significant changes in epigenetic aging were observed over the 4-week intervention. These findings demonstrate the feasibility and acceptability of incorporating epigenetic aging biomarkers into dietary intervention research and suggest that biological aging measures may serve not only as surrogate outcomes but also as tools to support participant engagement. The results also support metabolic syndrome as a relevant population for dietary geroscience trials and provide practical guidance for designing longer-term studies evaluating whether dietary interventions can slow biological aging and promote healthy longevity. ClinicalTrials.gov Identifier: NCT04361617 (date of registration: 04-23-2020).
Genotype-first screening compares patient genomes to a standard reference. The GRCh38 linear assembly contains reference minor alleles (RMAs), posing a critical structural vulnerability for automated clinical variant classification. To determine whether RMAs systematically generate false-positive annotations, we conducted an observational case series of 20 healthy adults undergoing preventive genome sequencing. Automated bioinformatic processing was performed using the GRCh38 linear reference to identify "high-impact" variant annotations generated at loci where GRCh38 differs from the population consensus major allele. Among all 20 participants (100%), linear alignment to GRCh38 systematically misclassified functional major alleles as false-positive "high-impact" annotations at three distinct loci (SLC37A4, CIMIP2A, and GPR33). These artifacts occurred solely because analysis software mathematically defined the healthy wild-type state as a deviation from the rare RMA. Cross-referencing with gnomAD confirmed these as population-dominant benign alleles. Furthermore, this inherent reference bias introduces a theoretical risk of false-negative classifications if RMAs mathematically mask true pathogenic variants. Automated pipelines using linear references systematically misclassify healthy alleles as high-impact functional annotations. While downstream population-frequency filters manage false-positive artifacts, this retrospective patching creates an unsustainable bottleneck for population-scale screening. Transitioning to graph-based pangenome references represents a highly promising approach to directly resolve these diagnostic vulnerabilities at the alignment level, though computational and standardization challenges must first be addressed to ensure the accuracy of large-scale aging research.
People with HIV (PWH) on effective antiretroviral therapy continue to experience a disproportionate burden of age-related comorbidities and multimorbidity. This review explores HIV-associated aging as a systems-level dysregulation and loss of resilience and evaluates emerging biomarker strategies for capturing its biological and clinical heterogeneity. Recent evidence indicates that aging in HIV is driven by interacting immune, metabolic, regenerative, structural, and inter-organ pathways that generate measurable signatures across biological scales. Composite biomarker approaches integrating molecular, physiological, and functional measures are increasingly being applied to cardiovascular, neurocognitive, physical, and co-infection-related outcomes. Large cohort studies, risk prediction models, and intervention trials support their clinical relevance, while emerging gerotherapeutic studies suggest that biologic aging markers may serve as responsive treatment endpoints. No single biomarker adequately captures HIV-associated aging. Future progress will depend on multimodal, outcome-anchored composites that integrate biological and functional domains and are coupled with artificial intelligence- and machine learning-based approaches to improve risk stratification, therapeutic monitoring, and evaluation of geroscience-informed interventions in PWH.
As global populations age and lifespan increases, health systems too often conceptualize "healthy aging" and "end-of-life care" as distinct phases. Yet emerging evidence from geroscience, palliative care, and health systems science suggests that aging and dying lie on a continuum of adaptation and care. This Perspective proposes a framework that treats system-supported, goal-aligned end-of-life care as a public health outcome of integrated life-course care and argues for system redesign to introduce earlier palliative approaches, align care models across the life-course, and reduce health disparities. This article outlines key mechanisms, evidence for integration, a conceptual model, and implications.
Epigenetic aging is a new dimensional advancement in geroscience that continually addresses factors beyond genetic determinants of longevity. It occurs through three conventional pathways, one of which is non-coding RNA production. miR-124 is a newly introduced miRNA that was found in aged skin and several tumorigenic cells and is associated with aging. Additionally, SIRT1, a protein from the NAD+-dependent family, is found to have a protective mechanism in the pathways of aging. On the other hand, vitamin D works as an epigenetic factor and often targets pathways related to the aging process. In this study, the aim is to identify the expression of miR-124 in circulating blood to address the epigenetic-related biomarker and link the facets with anti-aging SIRT1 and vitamin D cognate receptor (VDR) to understand vitamin D deficiency interception. A vitamin D deficiency middle-aged rat model was developed by assigning male Albino Wistar Rats. Paricalcitol was administered IP (intraperitoneal) to the experimental group (32 ng/day) to induce deficiency, along with a calcium-rich rescue diet. The study was conducted for 21 days following blood collection and storage. ELISA and RTqPCR were performed to estimate serum 25(OH)D and gene expression analysis, respectively. 25(OH)D comparison between the group p-value of 0.002. The log2 fold changes of SIRT1, VDR, and miR-124 were compared between the groups using boxplot visualisation. A linear regression model was developed to understand the explorative nature of aging index (R2 = 0.561; Adjusted R2 = 0.517; p = 0.005; n = 12). A putative epigenetic relation was found between miR-124 levels and their association with SIRT1. The investigated biomarkers of aging are associated with serum concentration of 25(OH) vitamin D in a middle-aged vitamin D-deficient rat model, which supports the need for mechanistic investigation in future studies.
Caveolin-1 (CAV1), the principal structural protein of caveolae, is a critical regulator of endothelial homeostasis, vascular aging, and frailty in older adults. By scaffolding multiple signaling pathways and controlling endothelial nitric oxide synthase activity, CAV1 shapes nitric oxide bioavailability, oxidative stress responses, and vascular stiffness. In parallel, extracellular vesicles (EVs) derived from endothelial cells, platelets, and other vascular cells act as key mediators of intercellular communication, propagating senescence, inflammation, and thrombosis across the vascular tree. This narrative review synthesizes current evidence on the CAV1-EV axis as a mechanistic nexus linking endothelial dysfunction, cardiovascular disease, and geriatric syndromes, with a particular focus on frailty. We first summarize the structure, regulation, and post-translational modifications of CAV1, especially tyrosine-14 (Tyr-14) phosphorylation, and their impact on endothelial senescence and dysfunction. We then examine how CAV1 controls EV biogenesis, cargo loading, and uptake, and how CAV1-enriched EVs contribute to pro-inflammatory and prothrombotic phenotypes in aging and frail populations. Finally, we discuss the translational potential of CAV1-related EVs as circulating biomarkers for early vascular aging and as targets for geroscience-guided interventions, including engineered or mesenchymal stem cell-derived EVs with vasculoprotective cargo. By integrating molecular, vascular, and geriatric perspectives, this review proposes the CAV1-EV axis as a promising framework for risk stratification and vascular-targeted therapies aimed at preserving function and independence in older adults.
Aging is commonly framed as a progressive accumulation of cellular and tissue-level damage. However, the aged organism does not decline as a collection of isolated organs. Aging is communicated systemically through blood-borne signals that connect senescent cells, immune remodeling, vascular dysfunction, metabolic stress, dysbiosis, and chronic inflammation. I propose the concept of the circulating senosome to describe the composite network of age-associated circulating mediators, including senescence-associated secretory phenotype proteins, extracellular vesicles, inflammatory cytokines, lipids, metabolites, complement and coagulation mediators, autoantibodies, cell-free nucleic acids, and microbiome-derived products. This framework positions blood as both a biomarker compartment and a therapeutic interface in aging biology. The circulating senosome does not replace established hallmarks of aging; rather, it provides a systemic layer through which multiple hallmarks interact. Defining, measuring, and therapeutically remodeling this circulating network may create new opportunities for translational geroscience.
Statin optimization in older adults is increasingly relevant within the geroscience framework of cardiovascular ageing. This randomized controlled trial evaluated whether pharmacist-led pharmaceutical care improves lipid outcomes and statin adherence in community-dwelling older adults receiving primary care. A total of 99 patients aged 65 years or older on long-term statin therapy were randomized to usual care or structured pharmacist-led care delivered every 3 months. The primary outcome was change in low-density lipoprotein cholesterol at 6 months. Secondary outcomes included changes in other lipid parameters and dispensing-based adherence assessed by the proportion of days covered. The present analysis represents a pre-specified interim evaluation and was conducted on a per-protocol population with complete 6-month laboratory data. No statistically significant between-group difference in low-density lipoprotein cholesterol reduction was observed (adjusted difference -0.129 mmol/L, p = 0.385). Statin adherence was high and stable in both groups throughout follow-up, with 84-87% of patients meeting adherence thresholds and approximately 75-77% achieving perfect dispensing, consistent with an apparent ceiling effect. Exploratory sex-stratified analyses identified potential differences in lipid responses, but these were not statistically robust. In these highly adherent elderly populations, pharmacist-led pharmaceutical care was not associated with additional short-term improvement in LDL-C or dispensing-based adherence. The findings highlight the importance of baseline adherence when evaluating adherence-focused interventions in older adults.
Vitamin D is increasingly recognized as a multisystem hormone whose actions converge on physiological domains central to frailty. Low circulating 25-hydroxyvitamin D [25(OH)D] concentrations are associated with sarcopenia, bone loss, immune and cognitive dysfunction, and excess mortality, while observational cohort studies link vitamin D deficiency to frailty onset and progression. Mechanistic evidence provides a biologically coherent framework for these associations. Vitamin D receptor (VDR) signaling regulates myogenic differentiation, neuromuscular junction integrity, osteoblast-osteoclast coupling, and osteocyte-mediated mechanotransduction. It also modulates innate and adaptive immune responses, restrains inflammasome activation, and preserves mitochondrial function and metabolic flexibility. Disruption of VDR signaling promotes muscle atrophy, skeletal fragility, immunosenescence, and impaired bioenergetic capacity, processes that mirror key biological features of frailty. Despite this strong mechanistic plausibility, randomized controlled trials have largely yielded neutral results in community-dwelling adults with adequate vitamin D status. Clinical benefits appear confined to individuals with deficiency or heightened vulnerability, whereas intermittent high-dose bolus regimens have been associated with increased risks of falls and fractures. These discordant findings reflect persistent methodological limitations, including heterogeneous frailty definitions, insufficient follow-up duration, inter-assay variability in 25(OH)D measurement, and uncertainty regarding thresholds relevant to non-skeletal outcomes. In this narrative review, we integrate clinical, epidemiological, and mechanistic evidence to position vitamin D as a biomarker and potential modulator of vulnerability within a broader geroscience framework.
Inflammaging, defined as the persistent, low-grade sterile inflammation accompanying aging, represents a central driver of age-related pathology, including cardiovascular dysfunction, neurodegeneration, metabolic disorders, and frailty. This review discusses the most recent advances in understanding its mechanistic basis, encompassing cellular senescence, the senescence-associated secretory phenotype (SASP), mitochondrial dysfunction, immune cell senescence, innate immune hyperactivation, defective inflammatory resolution, and nutrient-sensing dysregulation. Single-cell and spatial transcriptomics reveal tissue-specific and context-dependent patterns, highlighting the systemic complexity of inflammaging. Preclinical interventions demonstrate that inflammaging is modifiable through senolytics, which selectively eliminate senescent cells, and senomorphics, which suppress SASP without inducing cell death. Metabolic modulators such as metformin and rapamycin attenuate inflammatory signaling, while immune-directed therapies and microbiome-targeted interventions provide synergistic benefits through combinatorial approaches. Early-phase clinical trials in frail older adults show feasibility, safety, and preliminary efficacy, including reductions in circulating inflammatory markers, improved physical function, and enhanced immune responsiveness. Inflammaging trajectories are shaped by lifestyle, environmental exposures, and evolutionary factors, underscoring the need for personalized interventions. Remaining challenges include biomarker development, long-term safety evaluation, and heterogeneity across aging populations. Addressing these through interdisciplinary research supports a precision geroscience paradigm, where multimodal targeting of inflammaging can extend healthspan and reduce chronic disease burden.
Geriatricians are aware that the clinical presentation of older people is often with 'atypical' symptoms of commonly encountered conditions. In this Commentary, we explore the emergence of this description as geriatric medicine developed during the era that the disease-diagnosis paradigm dominated medical discourse. We suggest that this recognition of atypicality may have increased the sensibility of non-experts to the more subtle clinical complexity of unwell older adults and added legitimacy to the role of specialist geriatrics, and therefore contributed to improvements in the access and quality of care of older people. Then, noting the growing understanding of the multidimensional nature of illness causation, with age-related frailty now widely appreciated, and insights emerging from geroscience, we offer a critique of its conceptual limitations. Finally, we suggest that holding to the notion of atypicality might be an obstacle to a more assertive reorientation to equitable and age-attuned healthcare, and that the time may be right to offer atypicality a well-deserved retirement.
A central challenge in computational geroscience is to distinguish molecular quantities that predict mortality from those that causally drive it. Epigenetic clocks and aging biomarkers are increasingly used as if they were that mechanism, yet this is rarely tested directly. This distinction also bears on competing theories of aging: damage/reliability (A), hyperfunction/mTOR-IIS (B-1), and information loss (B-2). Although individual aging proteins have been tested piecemeal, no study has asked, in one framework, what fraction of mortality is measurable, whether it is causal, and whether it is reversible. Using only public, de-identified data, we evaluate this three ways. First, a Markov generator-matrix model of hallmark-load dynamics with death as an absorbing state, fitted by Bayesian inference through a joint biomarker-and-mortality likelihood to NHANES with linked mortality (n=23,844) and replicated in the Health and Retirement Study (HRS), decomposes Gompertz acceleration into visible (measured-biomarker-driven) and latent components. Second, a positive-control-calibrated, two-platform cis-pQTL Mendelian-randomization and colocalization design (UKB-PPP, deCODE) against parental-lifespan GWAS tests whether the latent's measurable components are causal. Third, a clock battery (Horvath, chronological; DamAge, causality-enriched damage) tests reversibility in cellular reprogramming. Within the model, ~92% of Gompertz acceleration is assigned to a latent component not captured by measured blood-biomarker axes (NHANES 92.5%, HRS 91.6%); the latent is partly encoded in DNA-methylation signatures but not transcription. The known causal proteins are detected (LPA p=9×10-12; IL6R p=2.8×10-5), yet the latent's components, across inflammatory, renal and growth-signalling (IGFBP3, IGF-1) axes, are null and do not colocalize on either platform. Reprogramming reverses the chronological clock (-11 to -22 yr) but not the causality-enriched damage clock. The model-inferred mortality-driving component is largely latent to accessible biomarkers; its measurable molecular proxies show no supported causal effect where the design detects known causes; and the causality-enriched damage-clock signal is resistant to partial reprogramming.
The convergence of neurological, psychiatric, neurodevelopmental, and public health approaches to brain health is reshaping global strategies for prevention, care, and policy. However, major gaps remain in the integration and implementation of brain and mental health frameworks across healthcare systems and regions. This paper describes the foundation and early development of the International Alliance on Brain Health (IABH), established in Switzerland in 2025 to promote interdisciplinary collaboration, reciprocal innovation, and implementation-oriented exchange between the global North and global South. Drawing on the Alliances founding meeting in Bern, its contribution as a partner to the World Brain Health Forum organized by the Paris Brain Institute, and its subsequent meeting in Buenos Aires, the paper outlines the Alliance's global positioning and priorities for translating brain health frameworks into practice. Key themes included integrated neurological and mental health approaches, prevention across the life course, digital innovation, brain capital, workforce development, and stronger inclusion of global South perspectives in international policy dialogue. The Alliance also emphasized bidirectional learning and locally adaptable implementation strategies aligned with WHO brain health frameworks. Under the joint patronage of the World Federation of Neurology and the World Psychiatric Association, the IABH represents an emerging platform for cross-continental collaboration in brain health.
We examined whether a genome-wide polygenic lifespan score (PLS) is associated with all-cause mortality and how this association compares with associations between long-term lifestyle factors and mortality. The PLS was computed for the older Finnish Twin Cohort (mean age 57.4 years; 45.2% men; N = 5575). Cox regression was used to estimate the effects of the PLS on all-cause mortality risk, before and after adding sex, physical activity, BMI, alcohol consumption, smoking behavior, and education level. Concordance indices (C-indices) were used to assess each predictor's contribution to the model's discriminatory performance. Over a mean follow-up of 17.5 ± 8.3 years, 1405 deaths (25.2%) occurred. A one standard deviation increase in the PLS was statistically significantly associated with a lower all-cause mortality risk (hazard ratio [HR] = 0.838, 95% confidence interval [CI] = 0.792-0.887). This association remained relatively unchanged after adding all covariates (HR = 0.863, 95% CI = 0.816-0.912). Smoking 20 or more cigarettes per day showed the strongest association with increased mortality risk (HR = 3.341, 95% CI = 2.751-4.056), while female sex was associated with the greatest risk reduction (HR = 0.678, 95% CI = 0.597-0.770). Smoking behavior had the largest impact on model performance (ΔC-index = 0.027); other covariates contributed less (ΔC-index < 0.006). In conclusion, genetic predisposition to a longer lifespan was associated with a modest reduction in all-cause mortality risk, independent of lifestyle and other factors. Smoking behavior and female sex were stronger predictors of mortality than the PLS.
Dementia is a major public health challenge, and Apolipoprotein E (APOE) ε4 is strongly associated with all-cause dementia, particularly Alzheimer's disease (AD). We aim to quantify the overall contribution of modifiable lifestyle, adiposity, socioeconomic status (SES), and health conditions occurring before dementia, to the association between ε4 genotype and the development of all-cause dementia, with AD examined as a major subtype. A population cohort study of 181,006 white UK Biobank participants aged ≥ 55 years at baseline was conducted, to examine the associations between APOE ε4 and all-cause dementia, and specifically AD, including modification and mediation role of lifestyle factors, adiposity, SES, and health conditions occurring before dementia. All risk factors, except for high alcohol intake, low diet quality, and phenotypic obesity, were associated with higher risk of all-cause dementia. The interaction contributions of lifestyle, adiposity, SES, and health conditions occurring before dementia varied by sex and dementia type. Low educational attainment had the strongest interaction effects with the association of APOE ε4 carriers and AD/all-cause dementia (up to 32.1%). In women, high deprivation level, abnormal sleep duration, anxiety, and depression showed interaction effects with APOE genotype (5-11.6%) as well. Phenotypic adiposity was associated with an increased risk of dementia among APOE ε4 non-carriers, but with a reduced risk among APOE ε4 carriers. Educational attainment explained a meaningful proportion of the APOE ε4 association with dementia. The strength of the association between APOE ε4 and dementia differed by risk factors and sex.
Evidence on meal timing and dementia is limited. We examined late-night dinner and skipping breakfast in relation to incident dementia. We conducted a population-based cohort study using linked medical insurance claims, long-term care insurance claims, and government-subsidized health checkup data from 19 participating in the Longevity Improvement & Fair Evidence (LIFE) Study. Cohort entry was from April 2016 to March 2021. Late-night dinner (dinner within 2 h of bedtime ≥ 3 times/week) and skipping breakfast (≥ 3 times/week) were assessed at health checkup and analyzed as dichotomous exposures (yes/no). Incident all-cause dementia was identified in claims data using ICD-10 codes. Associations were estimated using cause-specific Cox proportional hazards models, treating death as censoring and reporting hazard ratios with 95% confidence intervals, with prespecified adjustment for baseline demographic, lifestyle, and comorbidity factors. Among 283,166 participants (mean [standard deviation] age, 72.8 [5.5] years; 58.5% women), late-night dinner was reported by 36,609 (12.9%) and skipping breakfast by 17,727 (6.3%). During follow-up, 18,056 incident dementia cases and 16,390 deaths occurred. Both late-night dinner (hazard ratio, 1.17; 95% confidence interval, 1.12-1.20) and skipping breakfast (1.13; 1.06-1.20) were associated with higher hazards of incident all-cause dementia. Late-night dinner and skipping breakfast were associated with a higher risk of incident all-cause dementia in a large cohort of older Japanese adults. Meal timing behaviors assessed during routine health checkups may be relevant for dementia prevention strategies, although causal inference requires further study.
Understanding the links between metabolism, ageing, and age-related phenotypes may clarify the role of ageing in disease onset and improve risk prediction. We conducted a cross-cohort assessment of biological age using broad-spectrum LC-MS metabolomics of 3,686 plasma samples in 2,295 participants, aged 20-89, from the UK Airwave study (N = 960) and the Irish Longitudinal Study of Ageing (N = 1,335). The nucleoside N2,N2-dimethylguanosine, C-glycosyltryptophan, bile acid glucuronides, and the antioxidant zeta-carotene were associated with chronological age, frailty, and mortality. The noradrenergic metabolite 3-methoxy-4-hydroxyphenylglycol sulphate and the oligosaccharide sialyllactose were strongly associated with both age and mortality. We developed a metabolomic clock that was highly predictive of chronological age (r = 0.92) in test samples. Metabolomic age acceleration was strongly correlated between study visits (r > 0.6). Each standard deviation increase in metabolomic age acceleration (~ 5 years) was associated with 43% higher mortality risk, 27% higher risk of mild cognitive impairment, and 10% increased risk of a higher frailty score in fully adjusted models. The metabolites identified here may link ageing and age-related vulnerability and should be further investigated in mechanistic studies. The metabolomic clock has potential for translational applications, including as a prognostic and response marker of generalised age-related disease risk.
Fractures of long bones such as the femur are a common and serious health concern in the elderly, triggering immune responses essential for healing but also affecting remote organs like the liver. With age, the risk of fractures and immune imbalance increases, raising the likelihood of organ damage, infections, and mortality. To better understand age-dependent hepatic responses to bone injury, this study investigates early immune responses in the liver following femoral osteotomy, used here as a reproducible model of fracture. In a mouse model, young (17-26 weeks old) and aged (64-72 weeks old) male C57BL/6J mice received a femoral osteotomy with external fixation (Fx) or a corresponding sham procedure. After 24 h, inflammation, apoptosis, tissue damage, and immune responsiveness of the liver were analyzed. Aged sham animals exhibited a higher inflammatory state in sham-operated animals (neutrophil infiltration, tumor necrosis factor (TNF), interleukin (IL)-1b, chemokine (C-X-C motif) ligand 1 (CXCL1)), without corresponding increases in caspase-3-positive cells, activation of c-Jun N-terminal kinase (JNK), expression of sirtuin (SIRT) 1 or 3, or the receptor for advanced glycation end products (RAGE). Fx did not increase liver damage in young mice but showed a trend toward greater damage in aged mice. Fx significantly increased hepatic neutrophil infiltration and CXCL1 as well as TNF concentrations in both age groups. Aged mice showed weaker activation of pro- and anti-inflammatory signaling pathways after osteotomy, with reduced RAGE expression, JNK activation, and less pronounced induction of SIRT1 and SIRT3. In summary, femur osteotomy induced liver inflammation in both young and aged animals; however, older animals exhibit higher apoptosis and a superimposed inflammatory response on top of already elevated baseline inflammation in sham conditions. Further, they did not adequately activate key regulators like RAGE, SIRT1, and JNK, which coordinate inflammation and repair. Observed dysregulations may underlie the increased vulnerability to post-traumatic complications in the elderly. Understanding these age-related deficits is essential to improving therapeutic strategies.
Metabolic dysfunction-associated steatotic liver disease (MASLD) is a leading cause of hepatocellular carcinoma (HCC), particularly in obesity, yet mechanisms linking hepatocyte dysfunction to tumorigenesis remain unclear. Mixed lineage kinase domain-like protein (MLKL), the effector of necroptosis, is elevated in MASLD, but its hepatocyte-intrinsic role in obesity-driven MASLD-HCC is unknown. Using a long-term Western diet (WD)-induced MASLD-HCC model in hepatocyte-specific MLKL knockout (MlklHepKO) mice, we defined MLKL's hepatocyte-intrinsic function. WD increased hepatocyte MLKL protein expression without detectable necroptosis activation, indicating a necroptosis-independent role. MLKL deficiency did not alter WD-induced inflammation, fibrosis, or liver injury but increased hepatic lipid accumulation while reducing lipotoxic lipid species and preserving mitochondrial function. WD-fed MlklHepKO mice developed fewer and smaller tumors with reduced incidence, multiplicity, proliferation, and stemness. Transcriptomic analysis revealed upregulation of mitochondrial oxidative phosphorylation pathways in MlklHepKO livers. WD suppressed the mitochondrial fusion protein and tumor suppressor MFN2, whereas MLKL deficiency restored MFN2 expression post-translationally. In HCC cells, MLKL deletion reduced proliferation, improved mitochondrial respiration, and decreased glycolysis; these effects were reversed by MFN2 deletion. MLKL localized to nuclear and mitochondrial compartments, consistent with organelle-intrinsic functions. The human MLKL inhibitor necrosulfonamide (NSA) suppressed HepG2 xenograft growth, and elevated MLKL expression in human HCC correlated with poorer overall survival. Hepatocyte MLKL promotes MASLD-associated HCC through a non-necroptotic mechanism involving MFN2 suppression, impaired mitochondrial function, and increased tumor proliferation and stemness. These findings identify MLKL as a potential therapeutic target in MASLD-associated HCC.
Age-related endothelial dysfunction in the cerebral microcirculation contributes significantly to the pathogenesis of vascular cognitive impairment and dementia (VCID). Time-restricted eating (TRE) has emerged as a promising lifestyle intervention with beneficial effects on metabolic and vascular health; however, the mechanisms by which TRE influences the brain microvasculature remain incompletely understood. In particular, the role of circulating factors induced by TRE in modulating endothelial function has not been systematically investigated. Here, we tested the hypothesis that circulating factors derived from humans practicing time-restricted eating (TRE) induce protective and adaptive responses in human cerebromicrovascular endothelial cells. Using a serum transfer bioassay, endothelial cells were treated with serum obtained from aged individuals with or without TRE, followed by transcriptomic profiling. We demonstrate that TRE-associated serum elicits a robust and coordinated transcriptional reprogramming in human cerebromicrovascular endothelial cells, characterized by activation of stress-responsive and metabolic pathways and suppression of anabolic programs. Gene set enrichment analysis revealed significant activation of the integrated stress response (ISR)/ATF4 axis and suppression of mTORC1 signaling, consistent with a shift toward a catabolic, stress-adaptive state. These changes were accompanied by marked induction of the stress-responsive cytokine GDF15. At the mitochondrial level, TRE serum promoted increased expression of mitochondrial DNA-encoded oxidative phosphorylation components without activation of canonical mitochondrial biogenesis pathways, suggesting functional remodeling. Upstream regulator analysis identified coordinated activation of stress- and metabolism-associated transcription factors, including ATF4, FOXO, and KLF family members, alongside inhibition of anabolic regulators such as SREBF1/2. Notably, canonical endothelial functional programs, including autophagy and blood-brain barrier maintenance, were not coordinately activated. While individual angiogenesis-related genes were modestly upregulated, these changes did not translate into a coordinated pathway-level response. Collectively, these findings demonstrate that circulating factors induced by TRE promote a distinct endothelial phenotype characterized by metabolic reprogramming and stress adaptation rather than classical inflammatory or reparative responses. This work provides new mechanistic insight into how lifestyle interventions may influence cerebrovascular aging and identifies circulating factors as key mediators linking systemic metabolic state to endothelial function.