The impact of a polyphenol-rich supplement on epigenetic and cellular markers of immune age: a pilot clinical study

Abstract

Age-related alterations in immune function are believed to increase risk for a host of age-related diseases leading to premature death and disability. Programming of the immune system by diet, lifestyle, and environmental factors occurs across the lifespan and influences both makeup and function of the immune system, including immunometabolism. This programming is believed to act in large part through epigenetic modification. Among dietary components that affect this process, polyphenols may play an outsized role. Polyphenols are a widely distributed group of plant nutrients consumed by humans. Certain foods possess distinctive and relatively higher levels of these compounds. One such food is Tartary buckwheat (fagopyrum tataricum), an ancient seed historically prized for its health benefits. It is suggested that the specific composition of polyphenols found in foods like Tartary buckwheat may lead to a unique impact on immunometabolic physiological pathways that could be interrogated through epigenetic analyses. The objective of this study was to investigate the epigenetic effects on peripheral immune cells in healthy individuals of a standardized polyphenol concentrate based on naturally occurring nutrients in Tartary buckwheat. This pilot clinical trial tested the effects of consuming 90 days of this concentrate in 50 healthy male (40%) and female (60%) participants aged 18-85 years using epigenetic age clocks and deconvolution methods. Analysis revealed significant intervention-related changes in multiple epigenetic age clocks and immune markers as well as population-wide alterations in gene ontology (GO) pathways related to longevity and immunity. This study provides previously unidentified insights into the immune, longevity and epigenetic effects of consumption of polyphenol-rich plants and generates additional support for health interventions built around historically consumed plants like Tartary buckwheat while offering compelling opportunities for additional research.

Clinical trial registration: ClinicalTrials.gov, Identifier: NCT05234203.

Keywords: Tartary buckwheat; aging; diet and nutrition; epigenetic clocks; epigenome-wide association study; food-is-medicine; immunity; polyphenols.

Figure 1

Participant disposition chart.

Figure 2

Manhattan plots for the epigenome-wide association study (EWAS). The above plot depicts genes associated with CpG sites identified in the analysis. Each dot on the plot represents a CpG site, with its vertical position corresponding to the negative logarithm (base 10) of the unadjusted p-value for DNA methylation association, with a significance threshold set at p = 0.001. The x-axis shows genomic positions organized by chromosomes, with color-coded dots indicating specific chromosomes; different shades of blue are used to demarcate separate chromosomes. The prominently peaked dots represent CpG sites that surpass the genome-wide significance threshold, indicating significant associations.

Figure 3

Volcano plot for epigenome-wide association study and enrichment analysis. An epigenome-wide association study and enrichment analysis was conducted to compare pre- and post-intervention data (90 days after starting the study supplement). The Volcano plot illustrates differentially methylated loci (DMLs) identified in the pre- vs. post-intervention comparison. Each dot represents a CpG site, with its vertical position indicating the negative logarithm (base 10) of the unadjusted p-value for DNA methylation association. The x-axis shows the relative log fold change (logFC) of the m-values between the two timepoints. Negative values indicate CpGs with decreased methylation among study participants (green), while positive values indicate increased methylation (red).

Figure 4

Top 15 significant gene ontology terms associated with hypermethylated DMLs using GREAT. Gene ontology databases are reported for (A) GO-BP, (B) GO-MF, and (C) GO-CC. Biological associations shown are gene ontology (GO) terms for Biological Processes (BP), Molecular Function (MF), and cellular components (CC).

Figure 5

Top 15 significant gene ontology terms associated with hypomethylated DMLs using GREAT. Gene ontology databases are reported for (A) GO-BP, (B) GO-MF, and (C) GO-CC. Biological associations shown are gene ontology (GO) terms for Biological Processes (BP), Molecular Function (MF), and cellular components (CC).

Figure 6

Venn diagram of DMLs identified between the comparisons in (73) plotted against DMLs from the Tartary buckwheat supplement intervention.

Figure 7

Visual representation of individual epigenetic age accelerations (PCPhenoAge used as example). Each graph is faceted by each individual’s de-identified ID provided during the trial. Individual slopes calculated as changes in signify changes in epigenetic age metrics during this window (positive slopes represent increased epigenetic age acceleration, negative slopes represent decreased epigenetic age acceleration). These results may speak to individual differences in epigenetic sensitivity to environmental inputs, in this case dietary modification.