Metal mixtures and DNA methylation measures of biological aging in American Indian populations.

INTRODUCTION: Native American communities suffer disproportionately from elevated metal exposures and increased risk for cardiovascular diseases and diabetes. DNA methylation is a sensitive biomarker of aging-related processes and novel epigenetic-based "clocks" can be used to estimate accelerated biological aging that may underlie increased risk. Metals alter DNA methylation, yet little is known about their individual and combined impact on epigenetic age acceleration. Our objective was to investigate the associations of metals on several DNA methylation-based aging measures in the Strong Heart Study (SHS) cohort. METHODS: Blood DNA methylation data from 2,301 SHS participants was used to calculate age acceleration of epigenetic clocks (PhenoAge, GrimAge, DunedinPACE, Hannum, Horvath). Urinary metals [arsenic (As), cadmium (Cd), tungsten (W), zinc (Zn), selenium (Se), molybdenum (Mo)] were creatinine-adjusted and categorized into quartiles. We examined associations of individual metals through linear regression models and used Bayesian Kernel Machine Regression (BKMR) for the impact of the total metal mixture on epigenetic age acceleration. RESULTS: The mixture of nonessential metals (W, As, Cd) was associated with greater GrimAge acceleration and DunedinPACE, while the essential metal mixture (Se, Zn, Mo) was associated with lower epigenetic age acceleration. Cd was associated with increased epigenetic age acceleration across all clocks and BKMR analysis suggested nonlinear associations between Se and DunedinPACE, GrimAge, and PhenoAge acceleration. No interactions between individual metals were observed. The associations between Cd, Zn, and epigenetic age acceleration were greater in never smokers in comparison to current/former smokers. CONCLUSION: Nonessential metals were positively associated with greater epigenetic age acceleration, with strongest associations observed between Cd and DunedinPACE and GrimAge acceleration. In contrast, essential metals were associated with lower epigenetic aging. Examining the influence of metal mixtures on epigenetic age acceleration can provide insight into metals and aging-related diseases.

Molecular mechanisms of selenium-Induced spinal deformities in fish.

Selenium toxicity to oviparous vertebrates is often attributed to selenomethionine (SeMet), which can biomagnify through maternal transfer. Although oxidative stress is implicated in SeMet toxicity, knowledge gaps remain in how SeMet causes characteristic spinal deformities. In the present study, we use the Japanese medaka (Oryzias latipes) model to investigate the role of oxidative stress, cell death, and the unfolded protein response (UPR) on skeletal gene expression and SeMet toxicity, linking localization of cellular effects to observed abnormalities. Medaka embryos were treated with 2.5µM or 5µM SeMet for 24h at stage 25 (48h post fertilization). Post treatment, embryos were separated into normal, deformed (mild, moderate or severe), or dead categories. Dichlorofluorescein staining demonstrated oxidative stress in tails of embryos with observable spinal malformations. Furthermore, acridine orange staining for apoptosis identified significantly more dead cells in tails of treated embryos. Gene expression studies for the UPR suggest a potential role for CHOP (c/ebp homologous protein) induced apoptosis deformed embryos after 5µM SeMet, accompanied by a significant decrease in PDIA4 (protein disulfide isomerase A4) and no change in Dnajb9 (ER DNA J Domain-Containing Protein 4). This expression was distinct from the UPR induced by well-studied ER stress inducer, tunicamycin, which robustly activated CHOP, PDIA4 and Dnajb9. Finally, SeMet treatment significantly decreased transcripts of cartilage development, Sox9 (SRY box 9), while increasing Runx2 in deformed embryos, without altering Twist or Collagen 2a1. Results suggest that oxidative stress, the UPR and cell death play key roles in SeMet induced deformities and altered skeletal development factors.