A groundbreaking clinical trial shows we can reduce biological age (as measured by the Horvath 2013 DNAmAge clock) by more than three years in only eight weeks with diet and lifestyle through balancing DNA methylation.
A first-of-its-kind, peer-reviewed study provides scientific evidence that lifestyle and diet changes can deliver immediate and rapid reduction of our biological age. Since aging is the primary driver of chronic disease, this reduction has the power to help us live better, longer.
The study, released on April 12, utilized a randomized controlled clinical trial conducted among 43 healthy adult males between the ages of 50-72.
The 8-week treatment program included diet, sleep, exercise and relaxation guidance, and supplemental probiotics and phytonutrients, resulting in a statistically significant reduction of biological age–over three years younger, compared to controls.
The study was independently conducted by the Helfgott Research Institute, with laboratory assistance from Yale University Center for Genome Analysis, and the results independently analyzed at McGill University and the National University of Natural Medicine.
The study’s lead author, Kara Fitzgerald ND IFMCP, stated that “the combined intervention program was designed to target a specific biological mechanism called DNA methylation, and in particular the DNA methylation patterns that have been identified as highly predictive of biological age. We suspect that this focus was the reason for its remarkable impact.
“These early results appear to be consistent with, and greatly extend, the very few existing studies that have so far examined the potential for biological age reversal. And it is unique in its use of a safe, non-pharmaceutical dietary and lifestyle program, control group, and the extent of the age reduction. We are currently enrolling participants for a larger study which we expect will corroborate these findings.”
Leading epigeneticist Moshe Szyf PhD of McGill University and co-author on the study adds, “The uniqueness of Dr Fitzgerald approach is that her trial devised a natural but mechanistic driven strategy to target the methylation system of our body. This study provides the first insight into the possibility of using natural alterations to target epigenetic processes and improve our well being and perhaps even longevity and lifespan.”
DNA methylation patterns have become a leading means by which scientists evaluate and track biological aging, a term used to describe the accumulation of damage and loss of function to our cells, tissues and organs. This damage is what drives diseases of aging.
“What is extremely exciting,” commented Dr. Fitzgerald, “is that food and lifestyle practices, including specific nutrients and food compounds known to selectively alter DNA methylation, are able to have such an impact on those DNA methylation patterns we know predict aging and age-related disease.
I believe that this, together with new possibilities for us all to measure and track our DNA methylation age, will provide significant new opportunities for both scientists and consumers.”
The Effect of Diet on DNA Methylation in “Healthy” People
Beyond the efforts summarized above, other research groups focused their attention on the relationship between diet and DNA methylation before cancer or other diseases developed (Table 2). In 2010, Stidley and colleagues conducted a cross-sectional analysis on 1101 smokers from the Lovelace Smokers Cohort to evaluate the association of dietary factors with DNA methylation of eight genes (i.e., p16, MGMT, DAPK, RASSF1A, PAX5 α, PAX5 β, GATA4 and GATA5) in sputum samples.
Interestingly, they noted that high intake of green vegetables and folate was associated with lower DNA methylation, expressed as less than two genes methylated . A prospective analysis on the same cohort further demonstrated both positive (saturated fat) and negative (vitamin A, folate, and vitamin D) associations of dietary factors with DNA methylation of twelve cancer genes .
Summary of studies examining the relationship between dietary factors and DNA methylation in healthy people.
|First Author and Year of Publication||Country||Study Design||Study Population||Dietary Factors||DNA Methylation Markers||Sample Type||DNA Methylation Method||Main Findings|
|Stidley et al. 2010 ||USA||Cross-sectional||1101 smokers from the Lovelace Smokers Cohort||Total and animal fat, vitamin C, vitamin E, folate, carotene, alpha carotene, beta-carotene, lycopene, lutein and zeaxanthin, and retinol||p16, MGMT, DAPK, RASSF1A, PAX5 α, PAX5 β, GATA4 and GATA5||Sputum||Methylation Specific PCR||High intake of folate was associated with lower DNA methylation|
|Zhang et al. 2011 ||USA||Cross-sectional||161 cancer-free individuals||Folate, vitamins B12 and B6, riboflavin and methionine||LINE-1||Blood||Pyrosequencing||No association between intake of nutrients in one-carbon metabolism and LINE-1 methylation|
|Ono et al. 2012 ||Japan||Cross-sectional||384 healthy women||Folate and vitamins|
B2, B6, and B12
|Global DNA methylation||Blood||Methylight assay||Folate intake was negatively associated with global DNA methylation|
|Zhang et al. 2012 ||USA||Cross-sectional||180 cancer-free individuals||Folate and dietary patterns||IL-6 and LINE-1||Blood||Pyrosequencing||Folate intake was positively associated with LINE-1 methylation|
|Perng et al. 2014 ||USA||Cross-sectional||1002 participants of the Multi-Ethnic Study of Atherosclerosis study||Folate, vitamins B12 and B6, zinc, and methionine||LINE-1 and Alu||Blood||Pyrosequencing||Intake of methyl-donor micronutrients was not associated with DNA methylation|
|Agodi et al. 2015 ||Italy||Cross-sectional||177 healthy women||Mediterranean Diet and folate||LINE-1||Blood||Pyrosequencing||Women with low consumption of fruit and those with folate deficiency were more likely to exhibit LINE-1 hypomethylation|
|Shimazu et al. 2015 ||Japan||Cross-sectional||281 subjects without cancer and with no history of treatment against Helicobacter pylori infection||Green/yellow vegetables, fruit and salt||miR-124a-3, EMX1 and NKX6-1||Gastric mucosa||Methylation Specific PCR||Intake of green/yellow vegetables was negatively associated with methylation of miR-124a-3|
|Marques-Rocha et al. 2016 ||Brazil||Cross-sectional||156 subjects without metabolic disease, chronic inflammation, hydric balance disorders, changes in body composition and problems in nutrient absorption or metabolism||Energy and nutrients||LINE-1, TNF-α and IL-6||Blood||Methylation-sensitive high-resolution melting analysis||Individuals with higher LINE-1 methylation had higher daily intakes of calories, iron and riboflavin, and lower intakes of copper, niacin and thiamin|
|Nicodemus-Johnson et al. 2017 ||USA||Prospective||2148 Caucasian individuals from the Framingham Heart Study Offspring cohort||Fruits and juices||Genomic methylation profile||Blood||Infinium Illumina Human Methylation 450 k BeadChip arrays||There were 5221 and 5434 CpG sites associated with the intake of fruit and juice, respectively|
|Barchitta et al. 2018 ||Italy||Cross-sectional||299 healthy women||Mediterranean diet||LINE-1||Blood||Pyrosequencing||Adherence to the Mediterranean diet was positively associated with LINE-1 methylation level|
|Leng et al. 2018 ||Mexico||Prospective||327 Hispanics and 1502 non-Hispanic White smokers from the Lovelace Smokers Cohort||Nutrients||12 tumor suppressor genes||Sputum||Methylation Specific PCR||Intake of vitamin A, folate, and vitamin D was negatively associated with DNA methylation levels. Intake of saturated fat was positively associated with DNA methylation levels|
|Barchitta et al. 2019 ||Italy||Cross-sectional||349 healthy women||Foods and dietary patterns||LINE-1||Blood||Pyrosequencing||Consumption of whole-meal bread, cereals, fish, fruit, raw and cooked vegetables, legumes, soup, potatoes, fries, rice and pizza were positively correlated with LINE-1 methylation. LINE-1 methylation level increased with increasing adherence to a prudent dietary pattern|
|Mandaviya et al. 2019 ||Netherlands, Italy, Finland, USA, UK||Prospective||5841 participants with no history of cancer from 10 cohorts||Folate and vitamin B-12||Genomic methylation profile||Blood||Infinium Illumina Human Methylation 450 k BeadChip arrays||74 folate-associated DMRs, of which 73 were negatively associated with folate intake. The most significant folate-associated DMR was a 400-base pair (bp) spanning region annotated to the LGALS3BP gene|
|Zhang et al. 2011 ||USA||Cross-sectional||149 cancer-free individuals||Dietary patterns||LINE-1||Blood||Pyrosequencing||Adherence to a prudent dietary pattern was associated with a lower prevalence of LINE-1 hypomethylation. No association between the Western dietary pattern and LINE-1|
Several studies in this field of research evaluated the effect of dietary factors – and especially folate intake – on DNA methylation in blood samples. The epigenome-wide association study by Mandaviya and colleagues found 74 differentially methylated regions (DMRs) associated with folate intake, and, among them, the most significant was within the LGALS3BP gene .
Instead, evidence on global DNA methylation was controversial and characterized by positive, negative or no associations with folate intake. For instance, Ono and colleagues found a negative relationship between folate intake and global DNA methylation assessed by the luminometric methylation assay . Other studies, instead, used LINE-1 and/or Alu sequences as surrogate markers for estimating global DNA methylation level. While two studies failed in demonstrating any effect of nutrients in one-carbon metabolism on LINE-1 or Alu methylation [54,55], others found a positive association.
Interestingly, Zhang and colleagues demonstrated that folate intake from fortified foods was positively associated with LINE-1 methylation after adjusting for age, gender, race, BMI, diet, and physical activity . In line, Agodi and colleagues showed that women with folate deficiency, as well as those with low fruit consumption, were more likely to report LINE-1 hypomethylation .
The same research groups further demonstrated that adherence to specific dietary patterns might affect LINE-1 methylation in blood samples. In 2011, Zhang and colleagues suggested for the first time that adherence to a healthy dietary pattern was associated with a lower prevalence of LINE-1 hypomethylation . Consistently, Barchitta and colleagues first found that adherence to the Mediterranean diet was positively associated with LINE-1 methylation level , and then that this level increased with increasing adherence to a healthy dietary pattern .
As suggested by the authors, this relationship could be explained the consumption of healthy foods such as whole-meal cereals, fish, legumes, fruit and vegetables . This was partially in line with findings from the epigenome-wide association study by Nicodemus-Johnson and colleagues. Interestingly, they found more than 5000 CpG sites associated with the consumption of fruit and juice, respectively. Specifically, fruit-specific epigenetic signature could regulate genes associated with antigen presentation and chromosome or telomere maintenance, while the juice-specific epigenetic signatures were related to inflammatory pathways .
Only a few studies evaluated how energy consumption and intake of other nutrients affected DNA methylation globally. In particular, Marques-Rocha and colleagues showed that people with high intake of calories, iron and riboflavin, and those with low intake of copper, niacin and thiamin reported higher LINE-1 methylation than their counterparts. However, the authors failed in demonstrating any association of dietary factors with TNF-α and IL-6 methylation .
Finally, we selected the study by Shimazu and colleagues, evaluating the effect of fruits, green and yellow vegetables, and salt on DNA methylation of gastric mucosa of 281 subjects with no history of cancer and treatment against Helicobacter pylori infection. Specifically, the authors demonstrated that the intake of green and yellow vegetables was associated with lower methylation of miR-124a-3, but not with EMX1 and NKX6-1 genes .
The Relationship between Dietary Factors and DNA Methylation in Mothers and Their Children
Another field of application of studying the interaction between dietary factors and DNA methylation regards the effect of maternal diet on pregnancy outcomes and newborns’ health. To our knowledge, the first evidence of this relationships comes from a study on people who were prenatally exposed to famine during the Dutch Hunger Winter in the middle of the 20th century.
Indeed, sixty years later, study participants exhibited lower DNA methylation of the IGF2 gene when compared with unexposed individuals . Further investigations on the same cohort revealed additional DNA methylation changes in genes implicated in metabolic disorders, such as INSIGF2, GNASAS1, MEG3, IL-10 and LEP . More recently, findings from a genome-scale analysis confirmed that prenatal famine exposure was significantly associated with DNA methylation signatures in pathways related to growth and metabolism .
Accordingly, several observational studies evaluated the effect of dietary factors (i.e., nutrients, foods, and dietary patterns) on DNA methylation using data and sample from mother–child pairs (Table 4). In 2013, Boeke and colleagues failed in demonstrating an association of maternal intake of methyl donor nutrients with maternal and cord blood LINE-1 methylation .
Dietary cadmium, instead, was positively associated with maternal LINE-1 methylation at the first trimester of pregnancy, and negatively with cord blood methylation at birth . Consistently, the study by Taylor and colleagues did not find a significant effect of one-carbon metabolism nutrients on global DNA methylation in cord blood or buccal cells of children .
In contrast, Haggarty and colleagues demonstrated that folate intake and use of folic acid supplements were associated with low LINE-1 methylation in the offspring. The authors also observed that folate intake positively associated with IGF2 and negatively with PEG3 methylation .
With respect to IGF2, Rijlaarsdam and colleagues found that maternal diet high in fat and carbohydrates before pregnancy was positively associated with IGF2 methylation in offspring . Pauwels and colleagues evaluated the effect of maternal dietary factors before and during pregnancy on DNA methylation of RXRA, LEP, DNMT1, and IGF2. Interestingly, intake of betaine and methionine before pregnancy was positively associated with DNMT and LEP methylation; methyl group donor intake in the second trimester was negatively associated with LEP and DNMT methylation; intake of choline and folate in the third trimester was positively associated with DNMT methylation, and negatively with RXRA methylation .
To the best of our knowledge, only the study by McCullough and colleagues investigated the effect of complex dietary patterns rather than specific foods or nutrients. However, the authors demonstrated that a pro-inflammatory diets increased cytokine levels, but no effect on DNA methylation of nine genes was evident (i.e., IGF2, H19, MEG3, MEG3-IG, PEG3, MEST, SGCE/PEG10, NNAT, PLAGL1) .
Summary of studies examining the relationship between dietary factors and DNA methylation in pregnant women and their children.
|First Author and Year of Publication||Country||Study Design||Study Population||Dietary Factors||DNA Methylation Markers||Sample Type||DNA Methylation Method||Main Findings|
|Boeke et al. 2012 ||USA||Prospective||830 mother–child pairs||Vitamin B12, betaine, choline, folate, cadmium, zinc and iron||LINE-1||Maternal and infant cord blood||Pyrosequencing||No association of maternal intake of methyl donor nutrients with maternal and cord blood methylation. Periconceptional betaine intake was inversely associated with cord blood methylation; dietary cadmium was positively associated with first trimester methylation and inversely with cord blood methylation|
|Haggarty et al. 2013 ||United Kingdom||Prospective||913 mother–child pairs||Folate intake||PEG3, IGF2, small nuclear ribonucleoprotein polypeptide N, and LINE-1||Infant cord blood||Pyrosequencing||Folate intake was positively associated with IGF2 methylation and negatively with PEG3 and LINE-1 methylation in the offspring|
|McCullough et al. 2017 ||USA||Prospective||338 mother–child pairs from the NEST cohort||Dietary inflammatory potential||IGF2, H19, MEG3, MEG3-IG, PEG3, MEST, SGCE/PEG10, NNAT, PLAGL1||Infant cord blood||Pyrosequencing||Pro-inflammatory diets increased cytokine levels, but no association between dietary inflammatory potential and DNA methylation was evident|
|Pauwels et al. 2017 ||Belgium||Prospective||115 mother–child pairs from the Maternal Nutrition|
and Offspring’s Epigenome study
|Betaine, choline, folate, and methionine||Global DNA methylation and RXRA, LEP, DNMT1, and IGF2||Infant cord blood||Liquid chromatography–tandem mass spectrometry and Pyrosequencing||Before pregnancy, intakes of betaine and methionine were positively associated with DNMT and LEP methylation. In the second trimester, methyl group donor intake was negatively associated with LEP and DNMT methylation. In the last trimester, intake of choline and folate was positively associated with DNMT methylation and negatively with RXRA methylation|
|Rijlaarsdam et al. 2017 ||UK||Prospective||346 mother–child pairs from the Avon Longitudinal Study of Parents and Children||Dietary patterns||IGF2||Infant cord blood and blood at 7 years||Infinium Illumina Human Methylation 450 k BeadChip arrays||Maternal diet high in fat and carbohydrates before pregnancy was positively associated with IGF2 methylation at birth|
|Taylor et al. 2017 ||Australia||Prospective||73 children from the WATCH study||Methionine, folate, vitamins B2, B6 and B12 and choline||Global DNA methylation||Buccal cells||Enzyme-linked immunosorbent assay|
reference link :https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7466216/
Original Research: Closed access.
“Potential reversal of epigenetic age using a diet and lifestyle intervention: a pilot randomized clinical trial” by Kara Fitzgerald et al. Aging-US