Longevity Papers

Epigenetic clocks can predict biological age but cannot prescribe the interventions needed to reverse it. Here, we introduce REjuVenatIon Via Epigenetic Flow (REVIVE-Flow), a flow-matching model trained on a broad compendium of epigenetic blood studies to transport methylomes forward and backward in time. First, we learn the continuous vector field of aging as an Ordinary Differential Equation (ODE) within a stable, low-dimensional linear space. Then, the learned ODE\'s dynamics are integrated backward in time to define a natural, biologically-plausible rejuvenation trajectory. This path serves as a guide for a convex optimization problem that identifies the minimal, targeted CpG-level perturbation required to rejuvenate a sample. On a completely unseen test set comprising over 800 individuals from the European Prospective Investigation into Cancer and Nutrition (EPIC-Italy) cohort, Revive achieves 0.4 years rejuvenated per commanded year (R2=0.99), with a smooth sparsity-effect trade-off. Extensive validation confirms the effect preserves inferred cell-type composition and targets biologically plausible loci enriched in genomic regions and pathways central to aging biology.

Pulmonary Fibrosis (PF) is a life-threatening illness that is characterized by progressive scarring in the lung interstitium. There is an urgent need for new PF therapies because current treatments only slow down the progression of fibrosis, and the median life expectancy post-diagnosis is only 4-6 years. Since PF patients frequently exhibit telomere attrition, overexpressing telomerase, the enzyme responsible for synthesizing telomeres, represents a compelling therapeutic option. In this study, we in vitro transcribed human telomerase reverse transcriptase (hTERT) mRNA using modified nucleosides (modRNA). ModRNA hTERT treatment led to transient activation of telomerase activity in a dose-dependent manner in MRC-5 cells and, importantly, in primary human alveolar type II pneumocytes. Consequently, the proliferative capacity was increased, concomitant with reduced DNA damage and elongated telomere length. Notably, the induction of cellular immune response was only detectable at the highest modRNA concentration and returned to normal levels within 48 h. Next, we demonstrated that circularized, exonuclease-resistant modRNA hTERT extended the transient expression profile, which may be clinically advantageous. Finally, we provided therapeutic proof of concept in organotypic 3D ex vivo human precision-cut lung slices derived from end-stage PF patients. Intriguingly, a single modRNA hTERT treatment inhibited senescence, as indicated by significantly lower levels of senescence-associated β-galactosidase. Pro-inflammatory markers (IL6 and IL8) and, concurrently, the key fibrosis mediators TGFβ and COL1A1 were markedly reduced after modRNA and circular RNA hTERT treatment. In conclusion, the data presented herein provide initial evidence for the potential of RNA-based hTERT therapy for treating human lung fibrosis.

The transcription factor CCAAT/enhancer binding protein alpha (C/EBPα) regulates cell differentiation, proliferation, and function in various tissues, including the liver, adipose tissue, skin, lung, and hematopoietic system. Studies in rats, mice, humans, and chickens have shown that CEBPA mRNA undergoes alternative translation initiation, producing three C/EBPα isoforms. Two of these isoforms act as full-length transcription factors with N-terminal transactivation domains and a C-terminal dimerization and DNA-binding domain. The third isoform is an N-terminally truncated variant, translated from a downstream AUG codon. It competes with full-length isoforms for DNA binding, thereby antagonizing their activity. Expression of the truncated C/EBPα isoform depends on the initial translation of a short upstream open reading frame (uORF) in the CEBPA mRNA and subsequent re-initiation at a downstream AUG codon, a process stimulated by mTORC1 signaling. We investigated whether the ortholog of the CEBPA gene in the evolutionarily distant, short-lived African turquoise killifish (Nothobranchius furzeri) is regulated by similar mechanisms. Our findings reveal that the uORF-mediated regulation of C/EBPα isoform expression is conserved in killifish. Disruption of the uORF selectively eliminates the truncated isoform, leading to unrestrained activity of the full-length C/EBPα isoforms. This genetic modification significantly extended both the median and maximum lifespan and improved the healthspan of male N. furzeri. Furthermore, comparative transcriptome analysis revealed an upregulation of genes and pathways that are associated with healthspan and lifespan regulation in other species. These results highlight a conserved mechanism of CEBPA gene regulation across species and its potential role in modulating the lifespan and aging phenotypes.

The DREAM complex has emerged as a central repressor of DNA repair, raising questions as to whether such repression exerts long-term effects on human health. Here we establish that DREAM activity significantly impacts lifetime somatic mutation burden, and that such effects are linked to altered lifespan and age-related disease pathology. First, joint profiling of DREAM activity and somatic mutations across a single-cell atlas of 21 mouse tissues shows that cellular niches with lower DREAM activity have decreased mutation rates. Second, DREAM activity predicts the varied lifespans observed across 92 mammals, with low activity marking longer-lived species. Third, reduced DREAM activity in Alzheimer\'s patients predicts late disease onset and decreased risk for severe neuropathology. Finally, we show DREAM knockout protects against mutation accumulation in vivo, reducing single-base substitutions by 4.2% and insertion/deletions by 19.6% in brains of mice. These findings position DREAM as a key regulator of aging.

Loss of epigenetic information has been proposed as a potential driver of mammalian aging. However, its contribution to the well-documented variation in lifespan estimates among mammals remains to be elucidated. In this study, we examined DNA methylation entropy patterns at evolutionarily conserved CpG sites across multiple mammalian species to quantify age-associated epigenetic information loss. We found that longer-lived species tend to accumulate fewer CpGs exhibiting increased methylation noise over time, irrespective of whether these changes arise from hyper- or hypomethylation mechanisms. Importantly, the rate of epigenetic entropy gain declines in a linear fashion with species' maximum lifespan, pointing to the existence of a universal constraint on mammalian longevity, estimated to lie in the vicinity of 220 years. We further demonstrated that this relationship and its associated limit were independent of species and sample selection, as well as phylogenetic relatedness, and remained robust across different scenarios of lifespan estimation uncertainty. Collectively, this work highlights the maintenance of epigenetic information as a key factor in explaining lifespan differences among species and proposes a universal maximum limit to natural mammalian longevity.

The New England Centenarian Study (NECS) provides a unique resource for the study of extreme human longevity (EL). To gain insight into biological pathways related to EL, chronological age and survival, we used an untargeted serum metabolomic approach (> 1,400 metabolites) in 213 NECS participants, followed by integration of our findings with metabolomic data from four additional studies. Compared to their offspring and matched controls, EL individuals exhibited a distinct metabolic profile characterized by higher levels of primary and secondary bile acids - most notably chenodeoxycholic acid (CDCA) and lithocholic acid (LCA) - higher levels of biliverdin and bilirubin, and stable levels of selected steroids. Notably, elevated levels of both bile acids and steroids were associated with lower mortality. Several metabolites associated with age and survival were inversely associated with metabolite ratios related to NAD+ production and/or levels (tryptophan/kynurenine, cortisone/cortisol), gut bacterial metabolism (ergothioneine/ trimethylamine N-oxide, aspartate/quinolinate), and oxidative stress (methionine/methionine sulfoxide), implicating these pathways in aging and/or longevity. We further developed a metabolomic clock predictive of biological age, with age deviations significantly associated with mortality risk. Key metabolites predictive of biological aging, such as taurine and citrate, were not captured by traditional age analyses, pointing to their potential role as biomarkers for healthy aging. These results highlight metabolic pathways that may be targeted to promote metabolic resilience and healthy aging.

In the aging microenvironment, the decreased ability of bone regeneration seriously affects the efficiency of bone defect repair. To address this, a smart, reactive oxygen species (ROS)-responsive hydrogel-coated titanium implant loaded with copper-dihydromyricetin nanoparticles (CuDHM NPs) is developed. This implant synergistically modulates the bone repair microenvironment through dual mechanisms: anti-senescence and pro-angiogenesis. The hydrogel coating enables sustained, responsive release of CuDHM under oxidative stress conditions linked to cellular senescence. This mechanism effectively scavenges excessive intracellular and extracellular ROS accumulation, restores mitochondrial metabolic function, and directly decelerates the senescence of mesenchymal stem cells (MSCs). Moreover, the material induces the upregulation of key signaling molecules such as vascular endothelial growth factor (VEGF), promotes the formation of type H vessels, and synergistically ameliorates MSCs' senescence by modulating the extracellular matrix microenvironment. Notably, the formation of type H vessels itself enhances bone tissue regeneration. In vivo animal experiments demonstrate that the material accelerates bone repair by restoring local microenvironmental homeostasis and promoting vascularization. In summary, this study presents a novel implant that reprograms the microenvironment to combat age-related bone healing issues by addressing both cellular senescence and poor vascularization, offering a promising strategy for enhanced recovery in elderly patients with strong clinical potential.