Evaluation of epigenetic and metabolomic biomarkers indicating biological age

Biological age captures a person’s age-related risk of unfavorable outcomes using biophysiological information. Multivariate biological age measures include frailty scores and molecular biomarkers. These measures are often studied in isolation, but here we present a large-scale study comparing them. In two prospective cohorts ( n =3,196), we compared epigenetic (DNAm Horvath, DNAm Hannum, DNAm PhenoAge, DNAm GrimAge) and metabolomic-based (MetaboAge, MetaboHealth) biomarkers in reflection of biological age, as represented by five frailty measures and overall mortality. We observed that mortality-trained biological age markers, DNAm GrimAge and MetaboHealth, outperformed age-trained biomarkers in frailty reflection and mortality prediction. The associations of DNAm GrimAge and MetaboHealth with frailty and mortality were independent of each other and of the frailty score mimicking clinical geriatric assessment. Epigenetic, metabolomic, and clinical biological age markers seem to capture different aspects of aging. These findings suggest that mortality-trained molecular markers may provide novel phenotype reflecting biological age and strengthen current clinical geriatric health and well-being assessment. ### Competing Interest Statement The authors have declared no competing interest. ### Funding Statement The Rotterdam Study is supported by the Erasmus Medical Center and Erasmus University Rotterdam; the Netherlands Organization for the Health Research and Development (ZonMW); the Research Institute for Disease in the Elderly (RIDE); the Ministry of Education, Culture, and Science; the Ministry of Health, Welfare, and Sports; the European Commission; and the municipality of Rotterdam. The Leiden Longevity Study is supported by the European Union's Seventh Framework Programme (FP7/2007-2011) under grant agreement number 259679. This study was financially supported by the Innovation-Oriented Research Program on Genomics (SenterNovem IGE05007), the Centre for Medical Systems Biology, and the Netherlands Consortium for Healthy Ageing (grant 050-060-810), all in the framework of the Netherlands Genomics Initiative, Netherlands Organization for Scientific Research (NWO), by BBMRI-NL, a Research Infrastructure financed by the Dutch government (NWO 184.021.007 and 184.033.111). The current study was supported by VOILA (ZonMW 457001001) and Medical Delta (scientific program METABODELTA: Metabolomics for clinical advances in the Medical Delta). EBvdA is funded by a personal grant from the Dutch Research Council (NWO; VENI: 09150161810095). METD is funded by the Ministry of Health, Welfare, and Sport & The National Institute for Public Health of the Netherlands (S/010003). ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: Ethics committee/IRB of Leiden University Medical Center gave ethical approval for this work Ethics committee/IRB of Erasmus Medical Center gave ethical approval for this work I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines and uploaded the relevant EQUATOR Network research reporting checklist(s) and other pertinent material as supplementary files, if applicable. Yes Rotterdam Study data can be obtained upon request. Requests should be directed towards the management team of the Rotterdam Study (datamanagement.ergo{at}erasmusmc.nl), which has a protocol for approving data requests. For the data of the Leiden Longevity Study please contact Eline Slagboom or Marian Beekman (p.slagboom{at}lumc.nl) or (m.beekman{at}lumc.nl). Because of restrictions based on privacy regulations and informed consent of the participants, data cannot be made freely available in a public repository.