Fig. 1: LinAge2 predicts 20-year all-cause mortality and tracks with healthspan markers.

a–c Kaplan-Meier survival curves showing 20-year survival in the 65-74 CA bin (n = 631). For each clock, subjects were stratified by selecting the lowest (best, solid line) and highest (worst, dotted line) 25% quartiles for BA. Clocks within the same quartile were compared using log-rank tests with Benjamini-Hochberg correction. Areas shaded indicate 95% error bands for lines of the same color. b Compared to ChronAge, use of LinAge2 BA results in a significant survival difference for the lowest 25% BA quartile (P = 6.16E-04), but not for the highest 25% quartile (P = 0.07). PhenoAge Clinical did not significantly outperform ChronAge in predicting survival in this age bin. c LinAge2 significantly outperformed DunedinPoAm (P = 1.09E-02) and PhenoAge DNAm (P = 1.37E-03) in the lowest 25% BA quartile, but not GrimAge2 (P = 0.22). In the highest 25% quartile, while LinAge2 significantly outperformed PhenoAge DNAm (P = 0.03), the differences between LinAge2 and DunedinPoAm (P = 0.11) and GrimAge2 (P = 0.58) did not reach statistical significance. e ROC analysis revealed that LinAge2 (area under the curve (AUC) = 0.8684) was significantly more informative than PhenoAge Clinical (AUC = 0.8479, P = 6.35E-05) and ChronAge (AUC = 0.8288, P = 3.16E-10) in predicting future mortality (n = 2036). LinAge2 performed similarly to LinAge (AUC = 0.8647). f LinAge2 (AUC = 0.8440) also outperformed PhenoAge DNAm (AUC = 0.7859, P = 4.44E-07) and GrimAge2 (AUC = 0.8233, P = 0.016) in predicting 20-year mortality (n = 1,065). Although GrimAge2 outperformed ChronAge (AUC = 0.7933, P = 2.74E-03) in predicting 20-year mortality, PhenoAge DNAm did not (P = 0.47). a, d HorvathAge, HannumAge, and ChronAge did not significantly differ in predicting mortality risk (AUCs=0.7776, 0.7978, and 0.7933, respectively, n = 1065). ROC curves were compared using DeLong’s test. a, b, d–f CrystalAge, a theoretical perfect clock shown for reference, accurately identifies individuals at risk of dying (AUC = 1), whereas RandomAge adds random Gaussian noise of ±10 years to CA. g, h Violin plots for each clock categorized into low (biologically younger/best 25% quartile) and high (biologically older/worst 25% quartile) groups plotted against cognitive score (digit symbol substitution test) and gait speed. i–k Violin plots for each clock categorized by ability to perform (“yes” group) versus inability to perform (“no” group) employment work, all instrumental or all basic activities of daily living (iADLs and bADLs). Delta clock age refers to the age difference between an individual’s BA and CA. Groups were compared using two-sided t-tests. Median value, lower (25th) and upper (75th) percentiles are indicated. Lines extend to ±1.5 times interquartile range, with points outside this range drawn individually. The violin shape indicates the probability density function. yo years old, HA HorvathAge, LA2 LinAge2, GA2 GrimAge2, DPA DunedinPoAm.