1H-NMR metabolomics-guided DNA methylation mortality predictors

1H-NMR metabolomics and DNA methylation in blood are widely known biomarkers predicting age-related physiological decline and mortality yet exert mutually independent mortality and frailty signals. Leveraging multi-omics data in four Dutch population studies (N=5238) we investigated whether the mortality signal captured by 1H-NMR metabolomics could guide the construction of novel DNA methylation-based mortality predictors. Hence, we trained DNA methylation-based surrogates for 64 metabolomic analytes and found that analytes marking inflammation, fluid balance, or HDL/VLDL metabolism could be accurately reconstructed using DNA-methylation assays. Interestingly, a previously reported multi-analyte score indicating mortality risk (MetaboHealth) could also be accurately reconstructed. Sixteen of our derived surrogates, including the MetaboHealth surrogate, showed significant associations with mortality, independent of other relevant covariates. Finally, adding our novel surrogates to previously established DNA-methylation markers, such as GrimAge, showed significant improvement for predicting all-cause mortality, indicating that our metabolic analyte-derived surrogates potentially represent novel mortality signal. ### Competing Interest Statement The authors have declared no competing interest. ### Funding Statement This work was performed within the BBMRI Metabolomics Consortium funded by: BBMRI-NL (financed by NWO 184.021.007 and 184.033.111), X-omics (NWO 184.034.019), VOILA (ZonMW 457001001) and Medical Delta (METABODELTA: Metabolomics for clinical advances in the Medical Delta). EvdA is funded by a personal grant of the Dutch Research Council (NWO;VENI:09150161810095). Acknowledgements for all contributing studies can be found in the Supplementary Material-BIOS Consortium. Additional NTR samples were funded by the European Research Council (ERC-230374) project Genetics of Mental Illness (Boomsma). ### 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: Leiden Longevity Study (LLS) The Leiden Longevity Study protocol was approved by the Medical Ethical Committee of the Leiden University Medical Center before the start of the study (P01.113). In accordance with the Declaration of Helsinki, the Leiden Longevity Study obtained informed consent from all participants prior to their entering the study. LIFELINES-DEEP The LifeLines DEEP study was approved by the ethics committee of the University Medical Center Groningen, document number METC UMCG LLDEEP: M12.113965. All participants signed an informed consent form before study enrollment. All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Rotterdam Study The Rotterdam Study protocol was approved by the Medical Ethics Committee of the Erasmus MC Rotterdam, the Nethrlands. (MEC 02.1015) and by the Dutch Ministry of Health, Welfare, and Sport (Population Screening Act WBO, license number 1071272-159521-PG). In accordance with the Declaration of Helsinki, the Rotterdam Study obtained written informed consent from all participants prior to their entering the study. VUNTR The Netherlands twin Register study protocol was approved by the Central Ethics Committee on Research Involving Human Subjects of the VU University Medical Centre, Amsterdam, an Institutional Review Board certified by the U.S. Office of Human Research Protections (IRB number IRB00002991 under Federal-wide Assurance- FWA00017598. In accordance with the Declaration of Helsinki, the Netherlands Twin Register obtained informed consent from all participants prior to their entering the study. 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, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes All data produced in the present study are available upon reasonable request to BBMRI-NL and BIOS.