Longevity Papers

RNA splicing determines which protein isoforms a gene produces, and aberrant splice-site activation can cause severe disease. In Hutchinson-Gilford progeria syndrome, a synonymous LMNA mutation strengthens a cryptic 5' splice donor site to produce progerin, a toxic lamin A isoform driving premature cardiovascular death. Within this cryptic donor, the activating mutation lies near the canonical GT dinucleotide required for splice-site recognition. A single guide can therefore preferentially pair with the mutant sequence while positioning Cas9 cleavage at this dinucleotide. Here we show that CRISPR/Cas9-mediated disruption of this splice site suppresses progerin without requiring precise correction of the activating mutation and prolongs lifespan in progeroid mice. In patient-derived fibroblasts and induced pluripotent stem cell-derived mesenchymal stem cells, this intervention reduced progerin by 92.7-96.8% while retaining lamin A and lamin C expression. A single neonatal injection of AAV9 carrying the corresponding mouse guide increased median survival from 115 to 279 days in Cas9-expressing LmnaG609G/G609G mice. Treatment attenuated aortic, skeletal and multiorgan pathology. Treated homozygous pairs produced 52 offspring across seven litters. These findings establish pathogenic splice signals as genomic targets for sustained suppression of toxic isoforms while retaining canonical gene outputs.

Interest in fasting-based diets to improve metabolic health is growing. Caloric restriction (CR) with one meal per day includes an extended fasting component that contributes to its metabolic and longevity benefits, yet the role of fasting within CR remains unclear. Here, we compare CR with a fasting-refeeding-fasting (FRF) regimen while controlling food intake and fasting duration. Changes in plasma insulin and free fatty acids, hepatic mTOR signaling and ketogenesis, metabolic rhythms, and food digestion kinetics suggest that gastric emptying serves as a primary metabolic trigger in acute fasting. In contrast, CR fasting responses are regulated, suggesting anticipatory mechanisms. CR enhances circadian rhythmicity and metabolic gene coordination, whereas FRF disrupts it. CR improves glucose and fatty acid metabolism, while fasting leads to glucose intolerance and liver fat accumulation. These findings reveal that CR engages clock-aligned, anticipatory metabolic control, while fasting-refeeding cycles rely on direct nutrient cues.

Mild reductions in mitochondrial electron transport chain (ETC) capacity paradoxically extend organismal lifespan, a conserved phenomenon termed mitohormesis. While the mitochondrial unfolded protein response (UPRmt) and AMP-activated protein kinase (AMPK) are both established regulators of this process, whether distinct mitochondrial lesions converge on a single, unified survival mechanism remains unclear. By systematically dissecting the genetic architectures governing longevity in Caenorhabditis elegans, we demonstrate that targeted RNAi knockdown of Complex I (nuo-6) and Complex IV (cco-1) subunits activates fundamentally divergent downstream pathways. Both structural defects robustly induce the UPRmt, yet lifespan extension from Complex I impairment highly dependent on the UPRmt master regulator ATFS-1 while bypassing the AMPK ortholog AAK-2. Conversely, Complex IV-mediated longevity operates independent of ATFS-1 but requires AAK-2-driven metabolic reprogramming. Pharmacological intervention with metformin - a Complex I inhibitor and AMPK activator - further exposed complex-specific vulnerabilities: metformin markedly suppressed the longevity phenotype of nuo-6;aak-2 mutants, whereas in cco-1;aak-2 animals it produced a trend toward lifespan extension that appeared independent of AAK-2. Together, these findings challenge the view of mitohormesis as a uniform response, revealing instead that cells engage specialized, molecularly tailored retrograde signaling networks to govern lifespan and respond to pharmacological interventions.

Cellular senescence is a state of irreversible cell cycle arrest triggered by telomere erosion, persistent DNA damage or chronic stress. The accumulation of senescent cells disrupts tissue function and contributes to aging and disease. Here, we employ mass spectrometry-based proteomics to systematically interrogate dynamic proteome changes at multiple levels during the progression of replicative cellular senescence. We demonstrate that proteome changes during senescence occur in a coordinated manner, characterized by widespread protein depletion on chromatin. Moreover, components of the cytoplasmic translation machinery are depleted, while mitochondrial proteins display increased insolubility. Autophagic and proteasome activity is compromised in senescent cells along with remodeling of ubiquitin linkages and depletion of ubiquitin E3 ligases. Comparison of the senescent proteome with different pathophysiological cellular states reveals a distinctive senescent signature shaped by changes in the proteostasis network. Collectively, we provide a resource for the exploration of temporally resolved changes in the senescent proteome.

Existing aging theories describe what changes with age but do not prescribe how to intervene. We propose a control-theoretic framework that is not merely descriptive but prescriptive: it specifies which intervention, at which dose and sequence, under which safety constraints, will restore a measured biological state to a functional region. Aging is defined as progressive loss of safe controllability; biological age is the minimum safe control cost of functional restoration. Drugs are modeled as vector fields on biological state space whose non-commutativity, quantified by Lie brackets, predicts that intervention order determines outcome. The core differentiation from prior theories is operational: the framework outputs ranked targets, optimal sequences, safety-constrained protocols, and falsifiable predictions directly usable in drug discovery, rather than mechanistic ontologies or correlative biomarkers. We present a five-dimensional ODE model with analytic Lie-bracket derivation, a modality-aware control layer, three translational case studies, an implementation architecture with power analysis, and empirical scoring of aging interventions across five biological epochs. Twenty falsifiable predictions are enumerated. The central claim is that control-value reduction predicts translational success better than Hallmark annotation or biomarker reversal alone. If validated, this provides the missing interventional layer connecting aging biology to rational gerotherapeutic discovery.

Aging is a major risk factor for cardiovascular disease, but the role of the cardiac lymphatic vasculature in this process remains poorly understood. Here we show that aging reduces cardiac lymphatic vessel density in humans and mice and induces structural remodeling characterized by tighter, zipper-like endothelial junctions. These changes are associated with immune cell infiltration, fibrinogen and amyloid accumulation, and myocardial edema. Selective reduction of cardiac lymphatics in young mice, through Flt4 (VEGFR3) depletion or soluble Flt4 overexpression, recapitulates key features of cardiac aging, including inflammation and impaired lymphatic integrity. Mechanistically, aging induces selective upregulation of nuclear interleukin-33 (IL-33) in lymphatic endothelial cells. Unlike extracellular IL-33, nuclear IL-33 promotes lymphatic endothelial cell death and junctional remodeling. We identify VEGFC as an age-sensitive regulator that declines with aging and suppresses IL-33. Cardiac Vegfc overexpression or Il33 silencing restores lymphatic density and tissue homeostasis in aged hearts, identifying a potential therapeutic target for age-related cardiac dysfunction.

Aging is accompanied by molecular hallmarks conserved from yeast to humans. One such hallmark, senescence, is an irreversible nondividing state that promotes aging. We previously reported that mutants of the yeast Anaphase Promoting Complex (APC) shorten the lifespan of both dividing and nondividing cells. Here, we propose that activation of the APC will promote the maintenance of quiescent yeast cells, thereby delaying senescence and aging. We observed that APC activity in aging quiescent cells becomes increasingly impaired, as the APC substrates Clb1 and Mps1 accumulated within aging cells reintroduced back into the cell cycle. To identify peptide activators of the APC, we used a yeast 2-hybrid screen to recover peptides that interacted with the APC subunit Apc10. Recovered peptides were tested for their effects on the replicative lifespan (RLS) of dividing cells, and the chronological lifespan (CLS) of nondividing cells. Several peptides increased the RLS of wild type cells, but only one peptide (C43-4) increased CLS, which requires the APC Cdh1 co-activator, but not the Cdc20 co-activator. In cells expressing C43-4, Clb1 and Mps1 levels remained low when aging quiescent cells were reintroduced back into the cell cycle. The addition of a synthetic C43-4 to aging quiescent cells increased CLS even when added at late stages of aging. Mutations to APC subunits and co-activators blocked the ability of C43-4 to extend CLS. Htz1, which shares sequence homology with C43-4, was found to suppress APC mutant phenotypes and stabilize Apc10. We individually mutated the 6 amino acids in C43-4 that were shared with Htz1; one of the mutants disrupted the C43-4-Apc10 2-hybrid interaction and abolished the C43-4-dependent increase in CLS. C43-4 action is evolutionarily conserved, as C43-4 increased C. elegans lifespan in a daf-16- and aak-2-dependent manner. Our results describe the discovery of an evolutionarily conserved translational peptide that is the first of its kind to specifically activate the anti-aging APCCdh1 complex.

The proteasome is essential for proteostasis. Transcriptional induction of proteasomal components occurs when the proteasome is inhibited, but an overview of the transcriptional responses caused by proteasome perturbation is missing. Here, we profiled transcriptional changes caused by chemical and genetic proteasome inhibition and defined time-dose responses in cells and organoids. Induction of proteasome components varied by cell type and inhibition mode, whereas other responses were consistent, including upregulation of chaperones and secreted factors, and repression of cell cycle regulators. A proteasome stress response signature was defined based on the genes consistently modulated across systems, and applying this signature to aging datasets revealed activation of this stress response in some tissues, including skeletal muscle. Moreover, secreted factors within the signature showed similar age-related changes in human plasma, suggesting systemic activation of this stress response with aging. Together, these findings define a transcriptional signature for monitoring proteasome stress during aging and age-related diseases.

Glucagon-like peptide-1 receptor agonists (GLP-1 RAs) reduce major adverse cardiovascular events, all-cause mortality, and systemic inflammation in randomized controlled trials, with effect sizes exceeding those predicted from glycemic and weight-related improvements alone. The convergence of these findings with a maturing body of evidence linking metabolic dysfunction to accelerated epigenetic aging has prompted renewed interest in GLP-1 RAs as candidate gerotherapeutic agents. The present review synthesizes contemporary preclinical, mechanistic, and clinical evidence relevant to this question. The SELECT trial demonstrated a 19% reduction in all-cause mortality (hazard ratio [HR] 0.81) in patients with obesity without diabetes, the FLOW trial demonstrated a 24% reduction in the primary kidney composite endpoint (HR 0.76), and the first randomized evidence of GLP-1 RA modulation of validated DNA methylation clocks was reported in 2025, with significant deceleration of DunedinPACE, PCGrimAge, and PhenoAge over 32 weeks of semaglutide therapy. Mechanistic studies have identified convergent pathways involving the hypothalamic GLP-1 receptor, AMPK/SIRT1 signaling, and microbiome-derived short-chain fatty acid production. The aggregate evidence supports the framing of GLP-1 RAs as a candidate class of geroprotective therapeutics, although definitive trials with prespecified epigenetic aging endpoints, durability follow-up, body-composition assessment, prespecified sex-stratified analyses, and adequate representation of diverse populations remain to be conducted; the pending EVOKE and EVOKE+ readouts in early Alzheimer's disease will be particularly consequential.

Radiation-induced heart disease is a well-recognized complication of thoracic radiotherapy, with atrial fibrillation (AF) being a particularly notable sequela. The mechanisms by which ionizing radiation (IR) heightens susceptibility to AF remain inadequately understood. This study seeks to elucidate the role of cellular senescence and the GATA4-NF-κB signaling pathway in radiation-induced atrial remodeling and the pathogenesis of AF. An in vivo mouse model was developed through localized cardiac irradiation using 20 Gy X-rays. The irradiated mice demonstrated dose-dependent atrial structural and electrophysiological remodeling. Transcriptomic analysis revealed significant enrichment in DNA damage response (DDR), NF-κB signaling, and aging-related pathways. IR exposure induced substantial DNA damage, activated ATM/ATR pathways, and enhanced autophagic flux. These responses culminated in the accumulation of GATA4 and phosphorylation of NF-κB, which drove the expression of the senescence-associated secretory phenotype (SASP), coinciding with increased senescence-associated β-galactosidase (SA-β-gal) activity. In vitro, these crucial findings were recapitulated using irradiated HL-1 atrial cardiomyocytes, whereas the knockdown of GATA4 effectively suppressed both cellular senescence and SASP expression. Notably, treatment with the senolytic combination of dasatinib and quercetin (D/Q) alleviated DNA damage, inhibited excessive autophagy, and suppressed the GATA4-NF-κB pathway in irradiated mice, thereby reducing SASP levels. These improvements resulted in the reversal of atrial structural and electrophysiological remodeling and a marked reduced susceptibility to AF. Overall, our findings demonstrate that IR promotes atrial remodeling by inducing DNA damage-mediated cellular senescence via the GATA4-NF-κB-SASP axis. Targeting this pathway with senolytics such as D/Q thus identifies a promising therapeutic strategy for preventing radiation-induced atrial fibrillation.

Liver aging significantly impairs hepatic function and regenerative capacity, increasing the risk of morbidity and mortality from chronic liver diseases. Identifying molecular regulators of these processes may reveal promising therapeutic targets. Although Hepassocin (HPS), a hepatokine with known hepatoprotective functions, has minimal effects on liver homeostasis in adult mice, its role in long-term liver maintenance remains unclear. In this study, we observed a decrease in circulating and intrahepatic HPS levels in both aged mice and elderly humans. Moreover, the upregulation of HPS following two-thirds partial hepatectomy (PHx) was significantly blunted in 12-month-old (aged) mice. Aged HPS-knockout (KO) mice exhibited variable hepatic steatosis, exacerbated cellular senescence, and impaired autophagy. Liver regeneration after PHx was severely compromised in aged HPS-KO mice, as indicated by increased mortality, reduced hepatocyte proliferation, delayed liver mass recovery, and worsened autophagy disruption. Mechanistically, HPS directly activated 5'-AMP-activated protein kinase catalytic subunit alpha-1 (AMPK) in hepatocytes via the Annexin A2 (ANXA2)-extracellular signal-regulated kinase 2-90 kDa ribosomal protein S6 kinase 1-liver kinase B1 (ANXA2-ERK-p90RSK-LKB1) signaling cascade. Compared with their wild-type littermates, aged HPS-KO mice presented reduced LKB1 and AMPK activation and elevated mechanistic target of rapamycin kinase (mTOR) activity in both quiescent and regenerating livers. Treatment with the AMPK agonist AICAR ameliorated the liver aging phenotype and restored liver regenerative capacity in aged HPS-KO mice. Importantly, the administration of exogenous HPS enhanced regenerative outcomes in aged wild-type mice. These results establish HPS as a novel protective factor against liver senescence through AMPK-dependent mechanisms. Therapeutic strategies aimed at enhancing HPS signaling may offer a viable approach to counteract age-related liver dysfunction and regeneration failure.

Senescent cells are characterized by expression of markers like p16 and secretion of profibrotic and proinflammatory factors. The role of cellular senescence in age-related cardiac remodeling and dysfunction is incompletely understood. This study aimed to: (i) evaluate the effect of p16- positive cell clearance on cardiac function and structure in aging mice, and (ii) assess the role of different cardiac cell-types in the response. Hypertrophy markers, ion channels and calcium handling protein gene expression. Statistical analysis for all panels: one-way ANOVA followed by Tukey's test, significance level P<0.05 (N=6 for each group). Each point represents results from one mouse; bars and horizontal lines are means and SD. NK-ATTAC mice, permitting targeted clearance of p16-positive cells upon exposure to the dimerizing agent AP20187 (AP), were treated with AP or vehicle from 12 to 18 months of age. Cardiac function and structure were assessed with echocardiography, hemodynamics with a Millar catheter. p16-positive cells in various cardiac cell populations were analyzed with Fluorescence-Activated Cell Sorting (FACS) and immunofluorescence imaging. Echocardiography revealed significant attenuation of aging-associated increases in left ventricular mass to diameter at end-diastole (LVDd) and anterior wall thickness at end diastole (LVAWTd) in Aged-AP mice versus Aged-Vehicle. Diastolic dysfunction in vehicle mice normalized with AP treatment. FACS results indicated clearance of p16-positive fibroblasts with AP. Immunofluorescence imaging indicated reduced p16-positive fibroblasts and cardiomyocytes with AP, implicating them in the effects of p16-positive cell clearance on age-related cardiac remodeling. Exposure of cardiomyocytes to senescent fibroblast products led to upregulation of hypertrophy markers, pointing to paracrine effects on cardiomyocytes. This study highlights the potential contribution of senescent fibroblasts and cardiomyocytes to age-related cardiac remodeling. Modulating senescence might provide a new approach to age-related cardiac diseases like heart failure.