A Cost-Effective Saliva-Based Human Epigenetic Clock Using 10 CpG Sites Identified with the Illumina EPIC 850k Array

<b>Background/Objectives:</b> DNA methylation profiles have emerged as robust biomarkers of ageing, leading to the development of “epigenetic clocks” that estimate biological age. Most established clocks (e.g., Horvath’s 353-CpG pan-tissue clock and Hannum’s 71-CpG blood clock) require d...

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Main Authors: Christopher Collins, James Brown, Henry C. Chung
Format: Article
Language:English
Published: MDPI AG 2025-06-01
Series:DNA
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Online Access:https://www.mdpi.com/2673-8856/5/2/28
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author Christopher Collins
James Brown
Henry C. Chung
author_facet Christopher Collins
James Brown
Henry C. Chung
author_sort Christopher Collins
collection DOAJ
description <b>Background/Objectives:</b> DNA methylation profiles have emerged as robust biomarkers of ageing, leading to the development of “epigenetic clocks” that estimate biological age. Most established clocks (e.g., Horvath’s 353-CpG pan-tissue clock and Hannum’s 71-CpG blood clock) require dozens to hundreds of CpG sites. This study presents a novel saliva-specific epigenetic clock built on 10 sites identified from Illumina MethylationEPIC (850 k) array data. <b>Methods</b>: Saliva DNA methylation was analysed from 3408 individuals (age range 15–89 years, 68% male, 32% female, no diagnosed disease) from the Muhdo Health Ltd. dataset (2022–2024), and 10 CpG sites were selected where methylation levels showed the strongest positive correlations with chronological age (Pearson r = 0.48–0.66, <i>p</i> < 1 × 10−20). These CpGs map to genes involved in developmental and metabolic pathways (including <i>ELOVL2</i>, <i>CHGA</i>, <i>OTUD7A</i>, <i>PRLHR</i>, <i>ZYG11A</i>, and <i>GPR158</i>). A linear combination of the 10 methylation sites was used to calculate a “DNA methylation age”. <b>Results</b>: The 10-CpG clock’s predictions were highly correlated with chronological age (r = 0.80, R2 = 0.64), with a mean absolute error of ~5.5 years. Its performance, while slightly less precise than Horvath’s or Hannum’s multi-CpG clocks, is notable given the minimal marker set. It was observed that all 10 clock CpGs undergo age-related hypermethylation. The biological significance of these loci is discussed, along with the potential health and forensic applications of a saliva-based epigenetic age predictor. <b>Conclusions</b>: This study demonstrates that a saliva-specific epigenetic clock using only 10 CpG sites can capture a substantial portion of age-related DNA methylation changes, providing a cost-effective tool for age estimation.
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spelling doaj-art-a4be647817c544eab6f31564a4b6475e2025-08-20T03:27:28ZengMDPI AGDNA2673-88562025-06-01522810.3390/dna5020028A Cost-Effective Saliva-Based Human Epigenetic Clock Using 10 CpG Sites Identified with the Illumina EPIC 850k ArrayChristopher Collins0James Brown1Henry C. Chung2Muhdo Health Ltd., Columba House, Adastral Park, Martlesham Heath, Martlesham, Ipswich IP5 3RE, UKMuhdo Health Ltd., Columba House, Adastral Park, Martlesham Heath, Martlesham, Ipswich IP5 3RE, UKSchool of Sport, Rehabilitation and Exercise Sciences (SRES), University of Essex, Colchester CO4 3WA, UK<b>Background/Objectives:</b> DNA methylation profiles have emerged as robust biomarkers of ageing, leading to the development of “epigenetic clocks” that estimate biological age. Most established clocks (e.g., Horvath’s 353-CpG pan-tissue clock and Hannum’s 71-CpG blood clock) require dozens to hundreds of CpG sites. This study presents a novel saliva-specific epigenetic clock built on 10 sites identified from Illumina MethylationEPIC (850 k) array data. <b>Methods</b>: Saliva DNA methylation was analysed from 3408 individuals (age range 15–89 years, 68% male, 32% female, no diagnosed disease) from the Muhdo Health Ltd. dataset (2022–2024), and 10 CpG sites were selected where methylation levels showed the strongest positive correlations with chronological age (Pearson r = 0.48–0.66, <i>p</i> < 1 × 10−20). These CpGs map to genes involved in developmental and metabolic pathways (including <i>ELOVL2</i>, <i>CHGA</i>, <i>OTUD7A</i>, <i>PRLHR</i>, <i>ZYG11A</i>, and <i>GPR158</i>). A linear combination of the 10 methylation sites was used to calculate a “DNA methylation age”. <b>Results</b>: The 10-CpG clock’s predictions were highly correlated with chronological age (r = 0.80, R2 = 0.64), with a mean absolute error of ~5.5 years. Its performance, while slightly less precise than Horvath’s or Hannum’s multi-CpG clocks, is notable given the minimal marker set. It was observed that all 10 clock CpGs undergo age-related hypermethylation. The biological significance of these loci is discussed, along with the potential health and forensic applications of a saliva-based epigenetic age predictor. <b>Conclusions</b>: This study demonstrates that a saliva-specific epigenetic clock using only 10 CpG sites can capture a substantial portion of age-related DNA methylation changes, providing a cost-effective tool for age estimation.https://www.mdpi.com/2673-8856/5/2/28epigeneticsDNA methylationforensicsageing
spellingShingle Christopher Collins
James Brown
Henry C. Chung
A Cost-Effective Saliva-Based Human Epigenetic Clock Using 10 CpG Sites Identified with the Illumina EPIC 850k Array
DNA
epigenetics
DNA methylation
forensics
ageing
title A Cost-Effective Saliva-Based Human Epigenetic Clock Using 10 CpG Sites Identified with the Illumina EPIC 850k Array
title_full A Cost-Effective Saliva-Based Human Epigenetic Clock Using 10 CpG Sites Identified with the Illumina EPIC 850k Array
title_fullStr A Cost-Effective Saliva-Based Human Epigenetic Clock Using 10 CpG Sites Identified with the Illumina EPIC 850k Array
title_full_unstemmed A Cost-Effective Saliva-Based Human Epigenetic Clock Using 10 CpG Sites Identified with the Illumina EPIC 850k Array
title_short A Cost-Effective Saliva-Based Human Epigenetic Clock Using 10 CpG Sites Identified with the Illumina EPIC 850k Array
title_sort cost effective saliva based human epigenetic clock using 10 cpg sites identified with the illumina epic 850k array
topic epigenetics
DNA methylation
forensics
ageing
url https://www.mdpi.com/2673-8856/5/2/28
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