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Epigenetic regulation of aging: implications for interventions

of aging and diseases
Kang Wang1,2,3, Huicong Liu4, Qinchao Hu1,5,6,7, Lingna Wang4, Jiaqing Liu4, Zikai Zheng3,5, Weiqi Zhang3,5,8, Jie Ren3,5,8 ✉,
Fangfang Zhu4 ✉ and Guang-Hui Liu 1,3,8,9,10 ✉

Aging is accompanied by the decline of organismal functions and a series of prominent hallmarks, including genetic and epigenetic
alterations. These aging-associated epigenetic changes include DNA methylation, histone modification, chromatin remodeling,
non-coding RNA (ncRNA) regulation, and RNA modification, all of which participate in the regulation of the aging process, and
hence contribute to aging-related diseases. Therefore, understanding the epigenetic mechanisms in aging will provide new
avenues to develop strategies to delay aging. Indeed, aging interventions based on manipulating epigenetic mechanisms have led
to the alleviation of aging or the extension of the lifespan in animal models. Small molecule-based therapies and reprogramming
strategies that enable epigenetic rejuvenation have been developed for ameliorating or reversing aging-related conditions. In
addition, adopting health-promoting activities, such as caloric restriction, exercise, and calibrating circadian rhythm, has been
demonstrated to delay aging. Furthermore, various clinical trials for aging intervention are ongoing, providing more evidence of the
safety and efficacy of these therapies. Here, we review recent work on the epigenetic regulation of aging and outline the advances
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in intervention strategies for aging and age-associated diseases. A better understanding of the critical roles of epigenetics in the
aging process will lead to more clinical advances in the prevention of human aging and therapy of aging-related diseases.

Signal Transduction and Targeted Therapy (2022)7:374 ; https://doi.org/10.1038/s41392-022-01211-8

INTRODUCTION stress, deterioration of DNA repair and accumulation of DNA

Aging is a slow but gradual process that is characterized by a damage, changes in protein homeostasis leading to the accumu-
continuous decline in the normal physiological functions of living lation and aggregation of misfolded proteins, and changes in
organisms over their lifespan. With age, the body’s resilience epigenetic regulation. The word “epigenetics” is derived from the
decreases, making it more sensitive to aging-related diseases, Greek word “epi” and means “over” or “above” the genome.
such as neurodegenerative diseases and cancer, and increasing Epigenetics represents a reversible mechanism in regulating the
the risk of death.1–3 Although most organisms have a similar death function of the genome without altering the underlying DNA
curve and a higher mortality rate during aging, dramatic variance sequence of the genome; thus, the epigenome links genotype to
in aging rates can be observed within the same species. Taking phenotype, which plays an important role in modulating the aging
honey bees as an example, although the queen bee and the process in response to environmental stimulation.
worker bees are genetically identical, the queen lives on average Epigenetic modifications are often reversible with the aid of
ten times longer.4 Intriguingly, there are so-called “non-aging” epigenetic regulators, which lay the theoretical basis for aging
organisms that have exceptionally long lifespans, exhibit no or modulation and make them promising targets for aging-
late-onset aging-related declines in physiological abilities and are intervention strategies. However, it was not until recently that a
resistant to aging-related diseases, such as hydra and naked mole series of important studies have been carried out on epigenetic
rats.5–7 These phenomena indicate that aging is a complicated regulation and interventions for aging. In 1967, whole-genome
process that may be regulated by a variety of different factors. DNA methylation was found to be related to the age of spawning
Numerous studies have revealed how aging occurs and how it salmon.11 Subsequent studies revealed that DNA methylation was
is regulated by complex cellular and molecular mechanisms at generally downregulated in a variety of mouse tissues and human
different stages of life. Many factors affecting the aging process fibroblasts during aging.12,13 In 1987, the nucleosome occupancy
and longevity have been reported,8–10 including telomere short- in human skin fibroblasts was shown to decrease during aging,
ening, nutrient sensing, mitochondrial dysfunction and oxidative suggesting that chromatin configuration may change in the aging

1
State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, 100101 Beijing, China; 2State Key Laboratory of Stem Cell and Reproductive
Biology, Institute of Zoology, Chinese Academy of Sciences, 100101 Beijing, China; 3University of Chinese Academy of Sciences, 100049 Beijing, China; 4School of Biomedical
Engineering, Shanghai Jiao Tong University, 200030 Shanghai, China; 5CAS Key Laboratory of Genomic and Precision Medicine, Beijing Institute of Genomics, Chinese Academy of
Sciences and China National Center for Bioinformation, 100101 Beijing, China; 6Hospital of Stomatology, Sun Yat-sen University, 510060 Guangzhou, China; 7Guangdong
Provincial Key Laboratory of Stomatology, Guanghua School of Stomatology, Sun Yat-sen University, 510060 Guangzhou, China; 8Institute for Stem Cell and Regeneration,
Chinese Academy of Sciences, 100101 Beijing, China; 9Advanced Innovation Center for Human Brain Protection, National Clinical Research Center for Geriatric Disorders, Xuanwu
Hospital, Capital Medical University, 100053 Beijing, China and 10Beijing Institute for Stem Cell and Regenerative Medicine, 100101 Beijing, China
Correspondence: Jie Ren ([email protected]) or Fangfang Zhu ([email protected]) or Guang-Hui Liu ([email protected])
These authors contributed equally: Kang Wang, Huicong Liu, Qinchao Hu, Lingna Wang, Jiaqing Liu

Received: 30 June 2022 Revised: 14 September 2022 Accepted: 28 September 2022

© The Author(s) 2022

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Fig. 1 The history of studies on aging-associated epigenetic regulation and interventions

process.14 In 2010, histone methylation was first linked with life remodeling, RNA modification, and non-coding RNA regulation,
extension, and it was demonstrated that H3K4me3 demethylation is regulated during aging. We will also introduce current
in the germline boosted the lifespan of C. elegans.15 With therapeutic strategies to delay aging, including small molecules,
increasing epigenetic evidence related to aging, in 2013, the reprogramming, active health, and many other epigenetic-
concept of the “epigenetic clock” was proposed to link DNA associated approaches.
methylation with biological age.16 In addition, RNA modifications
and ncRNA regulation have recently been demonstrated to be
involved in the regulation of aging.17 Furthermore, emerging EPIGENETIC REGULATION OF AGING
single-cell chromatin modification profiling may provide molecu- Accumulating evidence from invertebrate and vertebrate organ-
lar information with an unprecedented resolution of the relation- isms, tissues, and in vitro systems links aging with epigenetic
ship between epigenetics and aging in the future.18–20 mechanisms. In mammals, there are global and local DNA
Understanding how aging is regulated by epigenetic factors methylation changes in the genome during aging. Additionally,
greatly facilitates the development of aging-delaying therapies. there is a general loss of histones as well as global chromatin
Ever since the first report of caloric restriction (CR) to slow down remodeling in all aging models. RNA modification and ncRNA
aging in 1935, researchers have been exploring potential regulation also play essential roles in cellular senescence via post-
approaches to delay aging.21 One of these aging-intervention transcriptional regulations. Studies on how these epigenetic
studies shows that the aging process can be delayed, and the mechanisms regulate individual aging can provide targets to
healthy lifespan or healthspan can be extended by CR and delay aging and rejuvenate aging organisms (Fig. 2).
lowering the basal metabolic rate.22 Another exemplary study is
the discovery of resveratrol, an agonist of the longevity factor SIR2 DNA methylation
of the sirtuin family, and its function in extending the lifespan of DNA methylation occurs at the cytosines in CpG dinucleotides to
yeast.23,24 In addition, heterochronic parabiosis (HP) and circadian form 5-methylcytosine (5-mC), and 60%-90% of CpG sites in the
rhythm models have been found to be effective in identifying mammalian genome are methylated (Fig. 3). The genome is
factors that delay aging.25,26 In 2011, senescent cells from generally hypomethylated during aging. Consistent with this,
centenarians or Hutchinson–Gilford progeria syndrome (HGPS) genes in energy metabolism and oxidative-stress resistance show
patients can be fully reprogrammed to a pluripotent state with a higher expression in skeletal muscle of aged individuals.32,33
rejuvenated epigenome, suggesting the potential of reprogram- DNA methyltransferases (DNMTs), namely DNMT1, DNMT3A,
ming in the reversal of aging.27,28 In 2015, a combination of and DNMT3B, add methyl groups to nucleotides, resulting in gene
dasatinib and quercetin was identified to kill senescent cells silencing.34,35 The expression of DNMT1 decreases with age,
selectively; hence they were named senolytic drugs.29 More resulting in a reduced DNA methylation level. DNMT1 mutants
recently, the concept of aging vaccines was proposed, and that cause the degeneration of selective central and peripheral
glycoprotein nonmetastatic melanoma protein (GPNMB) vaccina- neurons have been shown to translocate to the cytoplasm and
tion has been shown to decrease tissue senescence and alleviate form aggresomes while failing to bind to heterochromatin.36 In
aging-related phenotypes.30 However, how epigenetic mechan- contrast, the expression of DNMT3A and DNMT3B increases with
isms are involved in these aging-intervention approaches has just age, and contributes to de novo methylation of CpG islands in
begun to be revealed (Fig. 1).31 mammalian cells, increases with age.37,38 DNA methylation can be
In this review, we will discuss how epigenetic remodeling, removed by ten-eleven translocation (TET) enzymes.39 In clinical
including DNA methylation, histone modification, chromatin research of aged patients, mutations of TET2 or DNMT3A increase

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Fig. 2 An overview of the aging epigenome. During aging and the emergence of cellular senescence, a series of epigenetic changes occur in
cells, including alterations in DNA methylation, chromatin remodeling, histone modification, RNA modification, and ncRNA regulation

the expression of pro-inflammatory cytokines and chronic aVMPs and aDMPs, which can be measured to assess
inflammation, which is associated with conventional cardiovas- epigenetic age.
cular disease (CVD).40
Epigenetic clock
DNA methylation shift The level of CpG site methylation with age is a reliable biomarker
DNA methylation generally decreases with age in certain human to predict chronological age. Researchers have developed age
and mouse tissues or cell cultures.13,41,42,43 Compared with estimators called epigenetic clocks based on these mammalian
newborns, whole-genome DNA methylation in CD4+ T cells of DNA methylation levels. Epigenetic clocks use machine learning
individuals over 100 years old has been shown to be decreased.41 methods and are based on a set of CpG sites, whose DNA
The decrease of 5-mC from young to old mice is also observed in methylation states are consistent in multiple cells, tissues, or
various organs, such as the brain, liver, and small intestinal organs to predict the chronological age.51
mucosa, and the loss of 5-mC impairs the physiological function of The earliest model can estimate the age of a person and
cells in old mice.13 However, there is no notable shift in the predict the risk of aging-related diseases, but shows low
genome-wide methylation level during aging in other human cell precision, only explaining 73% of age variance with a prediction
types, such as cells of the epidermis, liver, and heart, or some rat error of 5.2 years.51 Since then, multiple epigenetic clocks have
tissues, such as blood and kidney. These differences in DNA been reported with higher accuracy, precision, and broader
methylation may be due to tissue specificity or different detection application prospects in aging research.52,53 Among them, the
techniques. On the other hand, many genes tend to be first-generation clocks are Horvath’s epigenetic clock and
hypermethylated at CpG islands with aging (Fig. 3).44–46 A large Hannum’s epigenetic clock.54,55 Horvath’s epigenetic clock is a
meta-analysis of aging-related CpG islands demonstrated that multi-tissue predictor based on 353 CpG sites to estimate the age
hypermethylation of CpG islands is conserved across 59 tissues, of most tissues and cell types and is widely used in aging and
including blood, liver, muscle, skin, brain, and cortex, derived from cancer research.16 Hannum’s epigenetic clock can measure and
128 mammalian species.47 compare human aging rates and provides a quantitative readout
Moreover, there are CpG sites that have increased variability in for aging-related diseases using 71 CpG markers from the DNA of
methylation with age, which are called age-associated variably blood.52 Based on Horvath’s pan-tissue clock, the DNAge™
methylated positions (aVMPs).44,48 Researchers first identified algorithm is developed to compare the chronological age of
aVMPs in twin studies, in which older monozygotic twins exhibit young and aged muscles.56 Later, the second-generation clocks,
a higher level of methylation variation in the overall content of including PhenoAge and GrimAge, introduced morbidity and
5-mC than younger twins, meaning that the methylation variation mortality into the model, improving accuracy over the first
increases with age.48 The increased variation in aVMP methylation generation.53,55 PhenoAge takes into account the role of multiple
is associated with the downregulation of the expression of clinical biomarkers and can predict 10-year and 20-year
pentose metabolism genes, including PYGL, TALDO1, and PGD.49 mortality.53 GrimAge is based on 12 plasma proteins and smoking
Apart from aVMPs, there are also specific CpG sites, named age- pack-years, and is a more predictive epigenetic clock for
associated differentially methylated positions (aDMPs).50 The identifying clinical phenotypes.55 Notably, a recent study built
methylation rate of aDMPs decreases with age in 6 mammalian up a single-cell age clock (scAge), which exhibits the epigenetic
species, including human beings, mice, dogs, naked mole rats, age using single-cell methylation data.57 ScAge is not only able to
rhesus macaques, and humpback whales.43 Thus, the DNA epigenetically differentiate “young” and “old” cells in hetero-
methylation shift is associated with different CpG sites, including geneous tissues, but also predicts the chronological age of the

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Fig. 3 The mechanism of DNA methylation and the epigenetic clock theory of aging. Aging is often marked by global DNA hypomethylation,
but hypermethylation also occurs at selective CpG islands. DNA methylation at the promoter of a gene often leads to silencing of that gene.
DNA methylation at the 5ʹ cytosine of CpG results in 5-methylcytosine (5-mC). The methylation of DNA is mediated by DNMTs whereas the
methyl group on DNA is removed by TET enzymes. TET enzymes oxidize 5-mC to generate 5-mC derivatives, including
5-hydroxymethylcytosine (5-hmC), 5-formylcytosine (5-fC), and 5-carboxylcytosine (5-caC), in mammalian cells. Age estimators, such as
Horvath’s clock, Hannum’s clock, PhenoAge, GrimAge and single-cell age clock (scAge) are based on DNA methylation changes in the genome

tissues in mice.57 In summary, different epigenetic clocks have Histone methylation

been developed for the prediction of the chronological age, Previous studies have shown that H3K4me3, a marker associated
which can be used to assess the efficacy of intervention methods with active transcription, plays an important role in determining
for aging and to advance precision medicine. aging and lifespan by regulating the expression of aging-related
genes.60,61 With aging in yeast, H3K4me3 accumulates in non-
Histone modification promoter regions and ribosomal DNA (rDNA), leading to the loss
Post-translational modifications of histones can activate or silence of rDNA heterochromatin along with an increase in genome-wide
gene expression and regulate the aging process. The types of pervasive transcription.62,63 Studies in C. elegans somatic cells
histone modifications include methylation, acetylation, phosphor- have also shown increased enrichment of H3K4me3 in promoter
ylation, ubiquitination, ADP ribosylation, and others.58 Among regions of senescence-related genes, and that this dynamics often
these modifications, methylation, and acetylation at lysine occurs in regions with relatively low H3K4me3 markers,64 while
residues are the most widely studied and are known to affect down-regulation of the ASH-2 trithorax complex leads to
the aging process. In vivo and in vitro studies report global H3K4me3 deficiency and lifespan extension.15 Consistently, ROS
changes in H3K9me3, H4K20me3, H3K27me3, and H3K9ac levels stimulation in C. elegans juveniles leads to an overall decrease in
during aging.59 Several enzymes are involved in the regulation of the H3K4me3 level and enhances their longevity.65 In a mouse
histone methylation and acetylation. Histone methyltransferases model of Alzheimer’s disease (AD), the level of H3K4me3 and its
(HMTs) and histone demethylases (HDMs) play opposite roles in catalyzing enzymes increases in the prefrontal cortex, a crucial
regulating histone methylation, and histone acetyltransferases brain region impaired in AD, and treating these mice with an
(HATs) and histone deacetylases (HDACs) antagonistically regulate inhibitor of H3K4 HMTs promotes the recovery of prefrontal cortex
histone acetylation (Fig. 4). functions.66 The H3K4me3 level has also been shown to increase

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Fig. 4 Chromatin structural remodeling during aging. A general loss of heterochromatin and detachment of lamina-associated domain (LAD)
structures from the nuclear lamina occur during this process. Higher-order chromatin structure alterations during aging are accompanied by
the redistribution of various histone modifications, including histone methylation (H3K4me3, H3K27me3, H3K36me3, H3K9me3) and
acetylation (H3K9ac, H3K56ac, H4K16ac, H3K18ac). This leads to reactivation of repeating sequences and dysregulated gene expression due to
aberrant chromatin accessibility

with age in mouse hematopoietic stem cells (HSCs).67 In contrast, somatic tissues of C. elegans, the global H3K9me3 level increases
a recent study using physiologically aged human HSCs demon- at heterochromatic regions in the distal arms of chromosomes,
strated that aging is associated with reduced H3K4me3, H3K4me1, but decreases in euchromatic central regions of autosomes.79 In
and H3K27ac.68 Neurons in aged (>60 years) human prefrontal aged Drosophila, H3K9me3 and HP1 signals on chromosomes are
cortex exhibit loss of H3K4me3 at 556 genes and gain of H3K4me3 significantly reduced compared with those in young flies, and
at 101 genes compared to young (<1 year) neurons.69 Thus, overexpression of HP1 extends lifespan.80 Additionally, diminished
H3K4me3 is related to aging in different species, although its levels of H3K9me3 and HP1 were identified in mesenchymal stem
influence on aging is context-dependent and requires further cells (MSCs) bearing pathogenic mutations of HGPS or Werner
investigation. Syndrome (WS), another human disease with accelerated
H3K27me3 is generally associated with gene silencing and aging.81–83 The expression of the H3K9me3 methyltransferase
compacted heterochromatin.70 Earlier studies suggested a global SUV39H1 is decreased during the aging of both human and
loss of H3K27me3 in aged C. elegans and prematurely aged cells mouse HSCs,84 leading to a global reduction in H3K9 trimethyla-
from Hutchinson-Guildford progeroid syndrome (HGPS) tion and perturbed heterochromatin function.84 Treatment of
patients,71 while in killifish and mouse brains, global H3K27me3 Werner syndrome (WS)-specific MSCs with vitamin C, gallic acid
increases with age.72 In C. elegans, the effect of the H3K27me3 (GA), or low-dose chloroquine (CQ) ameliorates a range of
demethylase UTX-1 on lifespan seems paradoxical, as both UTX-1 senescent phenotypes, promotes cell self-renewal, and upregu-
knockdown and overexpression in neurons and the intestine have lates levels of heterochromatin-associated marks, including
been shown to extend lifespan.73–75 The conserved histone lysine H3K9me3.85–87 Thus, a reduction or redistribution of H3K9me3 is
demethylases jmjd-1.2/PHF8 and jmjd-3.1/JMJD3 could work as observed across different species with aging, although this trend
positive regulators of lifespan in response to mitochondrial is also tissue and cell-type dependent.88,89
dysfunction across different species, suggesting that increased
levels of H3K27me3 in genes involved in the mitochondrial Histone acetylation
unfolded protein response (UPRmt) are detrimental to lifespan.76 Unlike histone methylation, the relationship between global
Determining the locus- and cell-type-specific roles of H3K27me3 histone acetylation and longevity is better understood. Histone
in lifespan regulation will be the key to unraveling the impact of acetylation is mediated by lysine acetyltransferases and is
H3K27me3-modifying enzymes on aging. increased in active gene regions. HDACs are considered to
H3K36me3 and H3K9me3 also play important roles in the aging function as corepressors and, together with HATs, play a critical
process. In both S. cerevisiae and C. elegans, deficiency of role in longevity. Sirtuins are class III HDAC that enhance genome
H3K36me3 is associated with a shorter lifespan. Consistently, the stability and regulate the deacetylation of lysine residues in an
loss of H3K36me3 demethylase extends the lifespan of S. NAD+ level-dependent manner.90,91 Among the sirtuin family
cerevisiae.77 Similarly, loss of H3K9me3 in the adult Drosophila members, SIRT1 has been reported to decrease with age in various
midgut leads to intestinal stem cell aging.78 Interestingly, in aged tissues of humans and mice, such as the liver, heart, kidney, brain,

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and lung.92,93 SIRT6 functions as an NAD+-dependent H3K9 is less clear, and more research is needed to refine the
deacetylase that modulates telomeric chromatin, and its over- mechanisms in the future.
expression contributes to the longevity of rat and human nucleus
pulposus cells via inhibiting senescence.94 On the other hand, the Chromatin remodeling
HDAC class II family member HDAC4 was reported to be Chromatin is a flexible and dynamic structure composed of DNA
polyubiquitylated and degraded during all types of senescence. and histones that can exist as heterochromatin or euchromatin.
HDAC4 selectively binds to and monitors H3K27ac levels at The basic unit of chromatin is the nucleosome core particle,
specific enhancers and super-enhancers, such as enhancers of encapsulated in a histone octamer consisting of a central H3-H4
AKR1E2 and VEGFC. Treatment with the inhibitor of the HAT P300 tetramer flanked by two H2A-H2B dimers.115 Chromatin remodel-
could rescue senescence in HDAC4-depleted cells, suggesting a ing is defined as a series of genome-wide changes in the nuclear
potential antagonistic effect between HDAC4 and P300.95 In architecture that can be recognized at the level of specific
IMR90 cells, P300, which promotes the formation of active chromosomes or chromosome domains, such as centromeres.
enhancer elements in the non-coding genome, significantly Significant chromatin structural remodeling has been identified
increases the levels of H3K122ac and H3K27ac at the proximal during cellular senescence, from histone component and mod-
senescence-specific gene promoters and is confirmed to be a ification changes to alterations of the chromatin compartments
primary driver of the senescent phenotypes, and depletion of and topologically associating domains (TADs).83,116,117 Global
p300 alone is sufficient to downregulate senescence genes and canonical histone loss is regarded as a common feature of aging
delay replicative senescence.96 CBP-1, the homolog of mammalian from yeast to humans.108,118,119 Overexpression of histone H3/H4
acetyltransferase CBP/p300 in C. elegans, is an essential regulator in yeast extends the lifespan, suggesting that an increased pool of
of the UPRmt and mediates H3K27ac and H3K18ac upon free histones promotes survival during aging by facilitating
mitochondrial stress. Knockdown of CBP-1 decreases the lifespan nucleosome exchange and post-transcriptional chromatin
of the worm in an HSF-1-dependent manner.97,98 Interestingly, repackaging.106 Genome-wide profiling of the core histone H3
abundant H3K4me1 marks are displayed in replicatively senescent occupancy in primary cultures of aging male mouse tissues and
IMR90 cells at the senescence-associated secretory phenotype neural stem cells (NSCs) reveals local changes in H3 occupancy as
(SASP)-associated super-enhancer loci, which also show substan- tissues and cells age, even though the H3 level remains relatively
tial H3K27ac marks and BRD4 binding.96,99 Recent studies also stable.120 Reversible phosphorylation of serine and threonine
demonstrated that aging-mediated changes in H3K27ac and residues in the C-terminal tail of H1 histones is responsible for
H3K9ac in the human cerebral cortex are associated with regulating the H1 stacking behavior. Individuals with mutations
AD.100,101 deleting these residues in one of the histone H1 isoforms show a
Owing to the antagonistic actions of HATs and HDACs, it is not progeria phenotype, and their fibroblasts exhibit more nucleoid
surprising that HATs have also been implicated in aging. The level relaxation, less condensed chromosomes, and higher nucleolar
of H3K14ac in the brains of aging mice can regulate the instability (Fig. 4).121
expression of aging-related synaptic plasticity genes, and the In eukaryotes, histone-modifying enzymes and ATP-dependent
H3K9me3/H3K14ac bivalent marks are significantly decreased in chromatin-remodeling complexes are the two main factors of the
old mouse hepatocytes.102,103 Inactivation of KAT7 decreases chromatin-remodeling process.122 Modified histones may induce
histone H3K14ac and alleviates human mesenchymal precursor conformational changes in nucleosomes. Restoration of acetyl
cell (hMPC) senescence.104 H3K18ac and H3K56ac are negative coenzyme A (acetyl-CoA) production through nutrient supple-
markers of senescence in Drosophila and yeast.105,106 Although the mentation (citrate, acetate, pyruvate, and glucose) could strongly
mechanism of action is different, knockdown of H3K56ac, Hst3, attenuate chromatin reorganization and diminish the extended
and Hst4-related HDAC-encoding genes during yeast aging lifespan of worms under mitochondrial stress conditions.123,124 In
shortens the lifespan.107 In aged yeast, the H3K56ac level mice, aged MSCs show significantly decreased levels of total
decreases while the H4K16ac level increases, leading to the histone H3-H4 acetylation and an increased abundance of
silencing of telomeric repeats.108 Interestingly, H4K16ac may also H3K27me3 across the gene body, resulting in a lower transcrip-
be involved in brain aging and AD progression. Normal aging tional rate and the loss of chromatin accessibility compared with
leads to H4K16ac enrichment, while H4K16ac in the proximity of young MSCs. Restoring cytoplasmic acetyl-CoA levels in aged
genes linked to aging and AD is dramatically reduced in AD.109 MSCs can remodel chromatin structure and rejuvenate these
Additionally, dysregulation of H4K12ac leads to aging-related cells.125 As H3K9me2 levels decrease, the nuclear peripheral
memory impairment, suggesting that it may serve as a critical heterochromatin loses its anchor to the nuclear lamina and moves
signal of memory formation.110 By administering the HDACi toward the nuclear interior.126 In specific regions during aging,
suberoylanilide hydroxamic acid (SAHA) to aged mice, the H3K9me2 switches to H3K9me3, another repressive mark but not
acetylation deficit of H4K12 could be rescued in neurons.111 Thus, enriched with direct contacts with the nuclear lamina; this may
substantial changes in histone acetylation occur during aging and reflect aging-associated changes in subnuclear location of
aging-related diseases, and understanding its regulatory mechan- peripheral chromatin and associate with shortened lifespan in
isms may provide new insight into the development of aging- aged C. elegans somatic tissues.79 Interestingly, histone deacety-
intervention strategies. lase or methyltransferase inhibitors alter histone modifications in
ways that predominantly increase euchromatin or decrease
Histone phosphorylation and ubiquitination heterochromatin.127 These results suggest that chromatin remo-
In addition to histone methylation and acetylation, histone deling is largely related to the level of histone post-translational
phosphorylation and ubiquitination have also been shown to be modifications. In addition, deletion of autophagy-related 7 (Atg7)
associated with aging, and in some cases through crosstalk with leads to disordered nucleosome assembly in mouse CD11b+Ly6G-
other histone marks. For example, the effect of histone bone marrow cells, resulting in cellular senescence.128 Promoters
ubiquitination on DNA damage accumulation can induce pre- of the conserved transcriptional and phenotypic responses to
mature neuronal aging.112 H3S28A mutants, which depletes defects in chromatin structure genes and are sensitive to histone
H3S28 phosphorylation but also reduces H3K27 methylation to dosage. Reducing nucleosome occupancy at these promoters by
prevent by compromising the activity of its methyltransferase deleting HHT1-HHF1 allows transcriptional activation induced by
complex in Drosophila, prolong lifespan and improve resistance the stress-responsive transcription factors Msn2 and Gis1, and
against starvation and paraquat-induced oxidative stress.113,114 thus, responses induced by moderate chromatin architectural
However, the correlation between these modifications and aging defects promote longevity.129

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ATP-dependent chromatin-remodeling complexes can be level.152 Although few studies have directly revealed the relation-
divided into the SWI/SNF, ISWI, CHD, and INO80 families. SWI/ ship between RNA modifications and organismal aging, accumu-
SNF is required for the activation of nutrient-responsive genes, lating data show the essential roles of these post-transcriptional
and the destruction of this complex impairs the ability of cells to regulatory mechanisms in the cellular senescence process, one of
adapt to their environment. SWI/SNF also regulates transcription the critical causes for aging and aging-related diseases (Fig. 5).
by remodeling chromatin and promoting a more open chromatin
configuration.130 In vitro, the BRM-SWI/SNF complex is required to m6A modification
promote co-expression of the telomere-binding proteins TRF1 and As one of the most extensively studied mRNA modifications in
TRF2, which are essential for maintaining telomere length and mammalian cells, m6A has been demonstrated to be involved in
structure in human fibroblasts and cervical cancer cells, contribut- cellular senescence. m6A is regulated by writer, reader, and eraser
ing to the development of longevity-related functions.131 SWI proteins.152,153 The multi-subunit writer RNA methyltransferases
plays a role in regulating aging during adulthood, and the absence (MTases) are assembled mainly by methyltransferase like 3
of SWI shortens the lifespan of nematodes.132 Among CHD (METTL3), METTL14, and Wilms tumor 1 associating protein
chromatin remodelers, the role of NuRD has been widely reported, (WTAP).154 The first reported m6A modification involved in
and disruption of the NuRD complex may compromise the senescence is methylation at the 3ʹ-UTR of the CDKN1A mRNAs
epigenetic composition of histones and the higher-order structure by a METTL3/14 heterodimer, which facilitates p21 translation.
of chromatin, making the NuRD complex more susceptible to the Consistently, the expression of METTL3/14 and p21 is enhanced in
influence of genotoxic stress.133 BAZ1A encodes an accessory oxidative-stress-induced senescence.17 Recently, ATG7 mRNAs
subunit of the ATP-dependent chromatin-remodeling complex with METTL3-dependent m6A were found to be destabilized by
that regulates cellular senescence in cancer and normal cells.134 the reader YTH N6-methyladenosine RNA-binding protein 2
Inhibition of the chromatin-remodeling factor SMARCA4 is able to (YTHDF2), which promotes senescence instead of autophagy in
prevent aging-dependent dopaminergic degeneration and short- fibroblast-like synoviocytes and leads to the progression of
ening of lifespan caused by α-synuclein and LRRK2 in Drosophila osteoarthritis.155 METTL14 also catalyzes the m6A modification
PD models.135 affecting miRNAs associated with senescence. For example, TNF-α-
Chromatin accessibility states and the expression programs of induced METTL14 overexpression leads to increased production of
aging-related genes are positively correlated during aging. Two miR-34a-5p from m A-modified primary transcript. miR-34a-5p
types of chromatin regions with regular changes in their promotes celluar senescence by targeting Sirtuin-1 (SIRT1) in
accessibility during aging are increased accessibility regions (IARs) nucleus pulposus cells (NPCs) of patients with intervertebral disc
and decreasing accessibility regions (DARs). IARs mainly exist in degeneration (IVDD), one of the most prevalent degenerative
genes related to the occurrence and development of aging, diseases.156 More recently, the regulator WTAP, which functions to
whereas DARs mainly exist in genes related to functional decline translocate METTL3/14 dimers to nuclear speckles,154 has also
caused by aging.136,137 The chromatin in human cells is spatially been demonstrated to be associated with IVDD. Increased WTAP
segregated into two compartments, compartment A and com- in senescent NPCs enhances the level of m6A in the lncRNA
partment B, and chromatin in the same compartment should have NORAD, contributing to the disruption of the NORAD/PUMILO/
more frequent interactions, as revealed by Hi-C analysis.138 The E2F3 axis and accelerating senescence.157
disruption of higher-order chromatin structure and the separation METTL3/14-mediated m6A modification has also been reported
of heterochromatin from the nuclear membrane are observed to inhibit senescence in some cases. METTL3/14 levels are
during cellular senescence and aging.63,83,130,139–141 A hierarchy of reduced in LMNA mutant-induced prematurely aged human
integrated structural state changes has been characterized HGPS cells and senescent fibroblasts, and METTL14 overexpres-
through large-scale epigenomic analyses of isogenic young, sion delays cellular senescence.158 The interaction of Lamin A and
senescent, and progeroid hMPCs, manifested as heterochromatin METTL3/14 protects the latter from proteasome-mediated degra-
loss in repressive compartments, euchromatin weakening in active dation to maintain sufficient m6A levels in normal cells.158
compartments, switching in interfacing topological compart- Moreover, METTL3-mediated m6A modification of MIS12 mRNAs
ments, and increasing epigenetic entropy. Nuclear lamina positively regulates their stabilization by recruiting IGF2BP2, and
dysfunction results in the derepression of constitutive hetero- in young hMSCs, MIS12 facilitates their self-renewal and alleviates
chromatic regions in repressive LAD structures marked by H3K9 cellular senescence. In HGPS and WS hMSCs, cellular models of
methylations. Diminished histone markers such as H3K27me3 and premature aging, the downregulation of MIS12 is detected at
facultative heterochromatin disruption contributed to an overall both the mRNA and protein levels.159 Knockdown of DNMT2 in
increase in epigenetic instability and ectopic expression of lineage mouse embryonic fibroblasts (MEFs) reduces the m6A level and
restricted genes, SASP genes and repetitive elements.83,142–145 accelerates senescence.160 Thus, in addition to RNA MTases,
Loss of heterochromatin leads to a global increase in transcription DNMT2 participates in senescence regulation by affecting the
and intercellular transcriptional heterogeneity, which is reported m6A level. Sulforaphane-mediated cycle arrest and senescence in
to be associated with cellular senescence and the onset of aging- breast cancer cells are also accompanied by downregulated
related diseases.81,137,143,146,147 Higher resolution data show that global m6A levels of mRNAs; however, the underlying mechanism
chromosome compartments are partitioned into TADs, which are is unclear.161
more basic domains with high interaction frequency therein and Similar to these m6A writers, the main erasers of m6A, i.e., fat
relatively isolated from neighbor regions. Below the scale of TADs, mass and obesity-associated protein (FTO) and alkB homolog 5
long-range chromatin looping interactions are insulated by TAD (ALKBH5), are also involved in aging. For example, the expression
boundaries.148 CTCF, a highly conserved architectural protein with of FTO declines with ovarian aging, followed by increased m6A
11 zinc fingers, functions as a barrier to inhibit heterochromatin levels in old human granulosa cells.162 Furthermore, FTO is crucial
spreading,149 and it has been shown to be reduced during aging for the progression of the G1 phase of the cell cycle by removing
in various models and plays an important role in chromatin m6A from the cyclin D1 mRNAs and stabilizing them.163 The
remodeling.150 Consistent with this, high levels of CTCF in ALKBH5 level is increased during IVDD and NPC senescence, and it
proliferating fibroblasts promote p16INK4a silencing.151 removes m6A from the DNMT3B mRNAs, which limits the
expression of the transcription factor E4F1 by methylating CpG
RNA modification islands at its promoter region and accelerates NPC senescence.164
More than 170 types of RNA modifications have been discovered To execute the function of m6A, reader proteins are needed,
thus far that regulate gene expression at the epitranscriptomic which include YTHDC family members, YTHDF family members,

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Fig. 5 RNA modifications involved in senescence. RNA modifications that have been revealed to be associated with senescence or aging
mainly include m6A modification, m5C modification, and adenosine-to-inosine (A-to-I) editing. The m6A modification is regulated by factors
including the writer RNA methyltransferases complex, the erasers FTO and ALKBH5, and the reader YTHDF2, whose effects on senescence are
complex based on different substrates. Under different stress conditions, m5C modification mediated by NSUN2 plays opposite roles in
senescence, retarding replicative senescence and accelerating oxidative-induced senescence. The A-to-I RNA editing catalyzed by the ADAR
family mainly exists in the central nervous system, and its relationship to neurodegenerative diseases has been demonstrated

the eukaryotic translation initiation factor eIF3, the insulin-like m5C modification
growth factor 2 mRNA- binding proteins (IGF2BP1/2/3), and the The m5C modification of RNAs is also tightly associated with
fragile X retardation protein (FMRP).165 Several studies have senescence, in which the diverse roles of the tRNA methyltrans-
indicated that the YTHDF family plays an important role in cellular ferase NSUN2 (NOP2/Sun domain family, member 2) depend on
senescence by destabilizing targeted mRNAs. In mouse embryonic the substrates of the m5C modification. For example, the 3ʹ-UTR of
stem cells (ESCs), m6A-modified intracisternal A-particle (IAP) cyclin‐dependent kinase 1 (CDK1) mRNAs and the 5ʹ-UTR of the
mRNAs recruit YTHDFs to shorten their half-life, repressing CDK inhibitor p27 mRNAs are targets of NSUN2, and m5C
endogenous retroviruses (ERVs).166 Conversely, the accumulation modification facilitates the translation of CDK1 while repressing
of IAP mRNAs after deletion of YTHDFs leads to high ERV that of p27, both of which alleviate replicative senescence.171,172
activity,166 resulting in senescence and diseases.167 The well- Interestingly, NSUN2 shows the opposite function in senescence
known senolytic therapy (discussed in the next chapter) of the under oxidative-stress conditions by targeting other mRNAs.
combination of dasatinib and quercetin can reduce the lipopo- NSUN2-mediated m5C at A988 in the 3ʹ-UTR of cyclin-dependent
lysaccharide (LPS)-induced SASP by upregulating YTHDF2, fol- kinase inhibitor 2A (CDKN2A, i.e., p16) mRNAs stabilizes the
lowed by destabilization of MAP2K4 and MAP4K4 mRNAs in human mRNAs, contributing to senescence.173 H2O2 treatment leads to
umbilical vein endothelial cells (HUVECs).168 upregulation of NSUN2 and Src homology 2 domain-containing
In addition to cellular senescence, the role of m6A methylation (SHC) family proteins in HUVECs with accelerated senescence,
in organs or organismal aging remains elusive. In brain aging and whereas knockdown of NSUN2 reduces ROS accumulation and
neurodegenerative disease models, dysregulation of m A and delays senescence. NSUN2 catalyzes m5C modification of SHC1
related regulatory proteins was indicated but the findings varied. mRNAs at several sites, which promotes its translation, activates
For example, a tendency toward an overall increase in m6A the p38 MAPK pathway, and leads to cell cycle arrest and elevated
methylation was observed in the cortex and hippocampus of the ROS levels.174 Strikingly, NSUN2 and METTL3/14 have synergistic
APP/PS1 transgenic mouse model for AD.169 Yet m6A in the 3ʹ UTR effects on the p21 mRNA methylation induced by oxidative stress.
of many AD-associated transcripts in the 5XFAD mouse model of The m5C modification induced by NSUN2 facilitates METTL3/14 to
AD is downregulated, which is accompanied by an 8% increase catalyze the m6A modification at the p21 mRNA 3ʹ-UTR and vice
and 4% decrease in FTO and METTL3, respectively, compared to versa.17 Furthermore, the RNA-binding protein human antigen R
wild-type mice, leading to higher Tau toxicity using the AD fly (HuR) facilitates the m5C modification at the C106 site of TERC to
model.170 enable telomerase activity and delay cellular senescence.175 In

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summary, these studies identify RNA m5C as an epitranscriptomic their relationship to organismal aging, which may provide an
marker for aging, and more investigation at the genome-wide avenue for developing new treatments for ameliorating
scale will further reveal the dynamics of the m5C landscape and its senescence.
potential impact on cellular senescence and tissue and
organismal aging. Non-coding RNA regulation
Research on the molecular mechanisms of cellular aging has
A-to-I editing mainly focused on protein-coding genes. However, accumulating
Adenosine to inosine (A-to-I) RNA editing conducted by the studies have demonstrated that ncRNAs, which widely regulate
adenosine deaminase acting on RNA (ADAR) family occurs most gene expression in multiple biological processes at the epigenetic,
frequently in the central nervous system, and the A-to-I imbalance transcriptional, and post-transcriptional levels, also play a critical
has been demonstrated to be involved in neurological disorders, role in aging. In recent years, studies of ncRNAs in aging have
metabolic diseases, and other diseases.176,177 Insufficient A-to-I mainly focused on microRNAs (miRNAs),180 long non-coding RNAs
editing influences neurodegenerative processes. The editing level (lncRNAs),181 R-loops,182,183 and circular RNAs (circRNAs)184 (Fig. 6).
at the GluA2 Q/R site in the hippocampal region is lower in AD, MiRNAs are small (~22 nucleotides), non-coding and single-
which induces altered Ca2+ influx and neuron death.178 In human stranded RNAs that bind to the 3ʹ-UTR of target mRNAs to degrade
endothelial cells under pro-inflammatory conditions, cathepsin S these mRNAs or suppress their translation.185,186 Using a micro-
(CTSS) mRNAs, encoding a cysteine protease, can be targeted by array containing 863 miRNAs, researchers discovered that 64
ADAR1, thereby recruiting HuR to improve its stability and miRNAs, such as miR-30d, miR-320d and miR-339-5p, are
translation. Consistently, the frequency of A-to-I editing on CTSS upregulated and 16 miRNAs, such as miR-103, miR-107, miR-24,
mRNAs is much higher in patients with vascular diseases.179 and miR-130a, are downregulated in long-lived individuals
Collectively, these reports indicate that RNA modifications play compared to younger individuals.180 In addition, the miRNA-p53
crucial roles in regulating senescence. However, further studies are pathway can maintain the genomic integrity in long-lived
needed to uncover more detailed underlying mechanisms and individuals during aging.180 The expression of miR-217 increases

Fig. 6 The mechanism of non-coding RNAs regulation during aging. Non-coding RNAs (ncRNAs) include microRNAs (miRNAs), long non-
coding RNAs (lncRNAs), R-loop (DNA-RNA hybrids), and circular RNAs (circRNAs). miRNAs bind to mRNAs, lncRNAs or circRNAs to prevent their
functions

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Fig. 7 The intervention of aging. Existing intervention strategies aim to alleviate aging in various organisms

in late-passage fibroblasts, where miR-217 inhibits DNMT1 cancer and neurodegeneration.183,196,197 For example, deletion
expression by targeting its 3′-UTR to induce human skin fibroblast of SPT6 extends lncRNA and increases R-loops associated with
senescence.187 The expression of age-associated miRNAs, includ- DNA damage, which ultimately leads to senescence in HeLa
ing miR-130, miR-138, and miR-181a/b, increases in keratinocytes cells.198 In summary, ncRNAs (miRNAs, lncRNAs, and circRNAs)
during cellular senescence, and by binding to p63 and Sirtuin-1 have been proven to serve as biomarkers in regulating cellular
mRNAs, these miRNAs affect cell proliferation pathways.188 senescence.
LncRNAs are non-protein-coding RNAs longer than 200 nucleo-
tides. LncRNAs bind to DNA, RNA, and proteins to exert their Strategies to alleviate aging
functions as guides, enhancers, or scaffolds in post-transcriptional Based on the molecular mechanisms underlying cellular senes-
and post-translational regulations.189 Therefore, lncRNAs have cence and aging, a series of therapeutic strategies, many of which
become targets for the treatment of fibrosis in aging. In aged are closely related to epigenetic regulations, have been proposed
bone marrow mesenchymal stromal cells, the lncRNA NEAT1 (Fig. 7). Reprogramming and geroprotective drugs have been
promotes CSF1 secretion and enhances osteoclastic differentia- developed to interfere with aging, while senolytics aim to remove
tion, which may be a therapeutic target for skeletal aging.190 The senescent cells to delay aging. Active health, such as caloric
lncRNA APTR accelerates the cell cycle and cell proliferation of restriction, exercise, and a healthy circadian rhythm, exerts
primary hepatic stellate cells in mice.191 Furthermore, targeting profound influences on multiple organs, systemic circuitries, and
the lncRNA Firre by CRISPR/Cas9 delays Ras-induced cellular whole-body rejuvenation. Moreover, several advanced interven-
senescence.192,193 tion methods have entered the clinical trial. Below we will discuss
Evidence shows that circRNAs play important roles in the all these aging-intervention strategies and their underlying
modulation of aging and aging-related diseases, such as epigenetic mechanisms.
cardiovascular disorders, diabetes, and neurodegenerative dis-
eases.194 As circRNAs are relatively stable, aging-related Small molecule-based therapy
increases in global circRNA levels are potential diagnostic The first class of aging-intervention strategies enumerated here is
biomarkers for aging.195 R-loops are three-stranded structures geroprotective drugs, which include epigenetic-related com-
composed of a DNA-RNA heteroduplex and a displaced single pounds (e.g., NAD+ precursors, sirtuin-activating compounds,
DNA strand. Although R-loops are often considered as “by- and HDAC inhibitors), small molecules with robust anti-diabetic
products” of transcription, recent studies have shown that effects (e.g., metformin), mTOR inhibitors (rapamycin), as well as
R-loops are important cellular regulators and may contribute to antioxidant chemicals (N-acetyl-l-cysteine) (Fig. 8).

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Fig. 8 Small molecule compounds as geroprotectors in diverse animal models. A series of small molecule compounds can extend the lifespan
or alleviate aging-related phenotypes in different organs. The interventions and corresponding target organs are shown in the diagram

NAD+ precursor insulin sensitivity and motor function and thus improves the
NAD+ is a critical redox coenzyme that plays a unique role in aging health and survival of mice on a high-calorie diet.211 Resveratrol is
through DNA repair and epigenetic regulation.199,200 The effect of also found to attenuate the aging of adipose stem cells via
sirtuins on histone deacetylation is highly dependent on NAD+, decreasing the levels of 5-mC in DNA and modulating mitochon-
highlighting the indispensable role of NAD+ in the epigenetic drial dynamics.212 SRT1720, an activator of sirtuin-1, can attenuate
regulation of aging.201 Supplementation with NAD+ precursors, vascular endothelial dysfunction, excessive superoxide produc-
such as nicotinamide mononucleotide (NMN), nicotinamide ribo- tion, aging-related metabolic diseases, and inflammation with
side (NR), and nicotinamide (NAM), prevents the decline in NAD+ aging, as well as improve the follicle pool reserve, thereby
and exhibits beneficial effects against aging and aging-related extending the lifespan and improving the healthspan of
diseases. NAD+ repletion extends the lifespan and delays the mice.213–215 SRT2104, another activator of sirtuin-1, preserves
accelerated aging in C. elegans and Drosophila melanogaster bone and muscle mass and extends the survival of male mice on a
models of Werner syndrome.202 In mammals, NR supplementation standard diet.216
increases mitochondrial function, delays the senescence of NSCs,
and increases mouse lifespan.203 NR supplementation also HDAC inhibitor
increases mitochondrial function and reduces aging-associated As discussed earlier, histone acetylation is one of the most
amyloidosis in muscle.204 In addition, NAD+ repletion with either important patterns of epigenetic regulation during aging. HDAC
NMN or NR ameliorates aging-associated meibomian gland inhibitors show geroprotective effects mainly through reversing
dysfunction in aged mice.205 It also improves cognitive functions aging-associated deacetylation of chromatin, acetylation of
in AD mouse models, mainly by rescuing cerebral microvascular histones near pro-longevity genes, and activating stress resistance
endothelial function and neurovascular coupling responses, and pro-longevity proteins.217 Administration of the pan-HDAC
preventing amyloid-β (Aβ) production in the brain, and reducing inhibitor SAHA rescues the skin phenotype, such as loss of
DNA damage, neuroinflammation, and apoptosis of hippocampal subcutaneous fat, inflammation, and fibrosis, in a mouse model of
neurons.206–208 NAM improves glucose homeostasis and reduces Cockayne syndrome (CS), a hereditary form of premature aging.218
hepatic steatosis and inflammation.209 Thus, boosting the NAD+ ITF2357 (givinostat) suppresses aging-induced diastolic dysfunc-
level appears to be a promising therapeutic strategy to counter tion in normotensive mice.219 Another HDAC inhibitor, butyrate
aging and aging-associated disorders, although its effects in protects against aging-related muscle atrophy in mice.220 HDAC
humans need further clinical studies. inhibitors also exhibit beneficial effects in neurodegenerative
disorders by modulating chromatin-mediated neuroplasticity and
Sirtuin-activating compound improving learning consolidation.221,222 Since sirtuins are also a
Activators of the sirtuin family of HDACs, also termed sirtuin- class of HDACs, the mechanism by which both STACs and HDAC
activating compounds (STACs), are another class of epigenetic inhibitors can delay aging remains to be further investigated.
drugs as potential geroprotectors. Since they were found to
promote the lifespan of yeast,24 STACs have been demonstrated Metformin
to extend the longevity of worms, fruit flies, honey bees, and Metformin is an anti-diabetic drug and one of the most attractive
fish.210 In mammals, resveratrol, an activator of sirtuin 2, increases geroprotective compounds, and it functions through extensive

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epigenetic regulation. Metformin retards aging in C. elegans by Reprogramming strategy
altering the ratio of S-adenosylmethionine (SAMe)/S-adenosylho- Reprogramming somatic cells with Yamanaka factors (Oct3/4,
mocysteine (SAH), which may affect histone methylation.223 In a Sox2, Klf4, and c-Myc; OSKM) reverses cell fate and finally
spatial restraint stress mouse model, metformin exerts antide- generates induced pluripotent stem cells (iPSCs), which possess
pressant effects by increasing the DNA 5-hmC modification level the characteristics of ESCs.288,289 A classic strategy for combating
of the Bdnf gene.224 Metformin treatment increases the microRNA- aging comes from the generation of iPSCs. The durable expression
processing protein DICER1 in mice and humans and thus modifies of OSKM leads to widespread chromatin remodeling,290 and
the profile of microRNAs associated with senescence and aging.225 interestingly, some aged somatic cells can be reprogrammed to
Administration of metformin also alleviates the senescence of exhibit a youthful state. Ectopic expression of OSK without c-Myc
dental pulp stem cells through AMPK/mTOR signaling pathway- restores the young patterns of DNA methylation and transcrip-
mediated downregulation of miR-34a-3p and upregulation of tomes in mouse retinal ganglion cells, which can ameliorate vision
CAB39.226 Strikingly, there is evidence that metformin intervention problems in glaucomatous in aged mice. The DNA demethylation
improves the lifespan and healthspan of mice even when the induced by OSK expression is confirmed to be necessary for the
administration starts at middle age (12 months)227 or old age rejuvenation process of retinal ganglion cells.291
(20–24 months),228–230 and the effect is enhanced when it starts Although long-term reprogramming rejuvenates aged cells to
earlier.231 In female SHR mice, however, metformin administration varying degrees, some of the aged somatic cells will be fully
starting at the age of 3 months increases the mean lifespan by reversed to iPSCs, which makes it impossible to be used to delay
14%, whereas the increase is only 6% when it starts at the age of aging in vivo due to the teratoma-forming ability of iPSCs.292
9 months, and there is no increase when it starts at the age of Notably, transient reprogramming, which allows the expression of
15 months. Consistent with this, the lifespan extension effect of reprogramming factors in a certain period of time, also exerts a
metformin is not seen in male rats232 or aged female mice.233 rejuvenating effect on aged somatic cells without altering the
Nevertheless, metformin relieves many aging-related diseases in original cell identities.293,294 Transfecting aged human fibroblasts,
rodent models, including cognitive impairment and neurodegen- chondrocytes, and endothelial cells with mRNAs expressing
eration,229,234–237 depression,238 chronic kidney disease,239 thymus OSKMLN (OSKM, LIN28 and NANOG) rejuvenates host cells and
degeneration,240 aging-related cataract,228 aging-related hearing significantly reverses the epigenetic clock.295 More recently, a 13-
loss,241 mitochondrial dysfunction in aged hearts,230 adipose day OSKM reprogramming using the Tet-on expression system
tissue senescence and metabolic abnormalities,242,243 and aging- significantly reduced the epigenetic age of human fibroblasts
related developmental and metabolic phenotypes.244 without fully changing them into iPSCs, indicating a boundary
between the rejuvenation and the pluripotency programs.296
Rapamycin Furthermore, short-term expression of OSKM in vivo significantly
Rapamycin, an approved immunosuppressant in solid organ expands the lifespan of progeria mice and restores the levels of
transplantation, also shows potential to intervene with aging. H3K9me3 and H4K20me3.292,297 In addition, a 2.5-week transient
Rapamycin extends the median and maximum lifespan of both reprogramming in early life (2-month-old mice) is sufficient to
male and female mice in a dose-dependent manner through extend the lifespan of transgenic progeria mice by 15% and
multiple mechanisms,245–247 including attenuating aging-related rejuvenates the DNA methylation patterns in skin cells.298 The
DNA methylation changes in the hippocampus to affect brain aging-associated epigenetic and transcriptional changes can also
aging,248 slowing the aging epigenetic signatures in mouse livers, be alleviated by transient reprogramming in naturally aged mice.299
and ameliorating a series of aging-related diseases including Overall, both long-term and transient reprogramming can achieve
cardiovascular dysfunction,249,250 neurodegeneration,251–254 ske- the rejuvenation of aged cells, while transient reprogramming also
letal muscle aging,255,256 ovarian aging,257,258 aging-related provides a novel method to alleviate aging in vivo in an organism.
hearing loss,259,260 and aging-associated periodontitis.261,262 How-
ever, prolonged rapamycin administration is reported to induce Senolytic therapy
muscle insulin resistance in rats, which might increase the Senolytics selectively clear senescent cells in aged individuals and
incidence of diabetes.263 Considering the immunosuppression have been studied as a potential therapy for aging intervention.
and NSC suppression effects of rapamycin,264 its application as a The first proposed senolytic strategy is the combination of
geroprotector should be assessed further. dasatinib (D) and quercetin (Q), two pan-tyrosine kinase inhibi-
tors.29 A single dose of D (5 mg/kg) + Q (50 mg/kg) effectively
N-acetyl-l-cysteine delays the aging phenotypes, such as frailty, cardiovascular
N-acetyl-l-cysteine (NAC) is an antioxidant with a prominent diseases, and IVDD in aged mice, and extends the lifespan of
influence on epigenetic regulation. It delays oocyte aging in mice Ercc1-/△ mice.29 To date, D + Q has been shown to prolong the
by increasing the expression of sirtuins.265 Similarly, NAC healthspan and the physiological or pathological aging process in
attenuates aging-related oxidative damage and neurodegenera- a variety of tissues or organs, including the cardiovascular
tion in rat brains by upregulating sirtuin-1 and downregulating system,300,301 skeleton,302–304 brain,305,306 adipose,307,308
several SASP factors (TNF-α, IL-1β, IL-6).266 In addition, NAC extends lung, 309,310
and muscle. 311
Most recently, epigenetics regulation
the lifespan of mice267 and ameliorates a series of aging-related has been demonstrated to be an important mechanism by which
diseases in rodents, such as AD,268,269 aortic fibrosis,270 immuno- D + Q eliminates aging cells. D + Q treatment leads to a
senescence,271 oxidative stress and senescence in the lung,272 significant change in epigenetic signatures in the hippocampus
bone loss in ovariectomized mice,273 adipose tissue senescence and improves the cognitive ability of aged male Wistar rats.312
and metabolic abnormalities,243 and aging-related hearing loss.274 Moreover, senescent adipose precursor cells exhibit hypomethyla-
tion and upregulated expression of the ZMAT3 gene, which is
Other geroprotective drugs related to type 2 diabetes; 3 days of D + Q treatment is able to
Many other drugs also show geroprotective effects, including anti- increase DNA methylation of ZMAT3 and decrease its expression,
diabetic drugs (sodium-glucose cotransporter-2 inhibitors,275 and reverse the senescence signature.313 In addition to D + Q,
acarbose,276–278) natural compounds (gallic acid,86 querce- other senolytic drugs such as ABT-263, ABT-737, digoxin, FOXO4-
tin,279,280) antioxidant molecules (vitamin C,85,281 methylene DRI (D-retro inverso), and heat shock protein (HSP) 90 inhibitor 17-
blue,282) antihypertensive drugs (angiotensin-converting enzyme DMAG, play their senolytic roles mainly by inducing apoptosis and
inhibitors and angiotensin receptor blockers),283,284 chloro- mitochondrial dysfunction,314–318 but their relationship to epige-
quine,87,285 aspirin,286 uridine,287 and so on. netics needs further investigation.

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Fig. 9 A healthy lifestyle to postpone aging. Active health, including caloric restriction, rhythm control and exercise, improves body function
and affects the lifespan of various animals, suggesting that healthy lifestyles exert profound effects on aging intervention

Active health intervention humans, which will be discussed in the ‘Clinical intervention’
Active health refers to choosing a healthy lifestyle autonomously, section later.
such as caloric restriction, regular routine and moderate exercise,
which are considered to benefit the quality of life and may exert a Circadian rhythm
rejuvenating effect on the aging process (Fig. 9). With the The circadian rhythm coordinates the behavior with the day/night
increasing awareness of active health, various studies have shift and is also considered to play an essential role in the aging
demonstrated that healthy lifestyles ameliorate aging-associated process.337 For example, epidermal and muscle stem cells from
features in different animals and humans. aged mice exhibit changed daily rhythms that cope with the stress
of aging environments, indicating the continuous change of
Caloric restriction circadian rhythms along with the aging process.338 Disturbed
CR, which reduces calorie intake ranging from 10 to 40%, has been circadian rhythm is linked to changes in chromatin structure, and
demonstrated to expand the lifespan of rodents to varying it is found that 6 h sleep deprivation affects the chromatin
degrees,319–321 attenuating vascular endothelial dysfunction, accessibility in the cerebral cortex of mice, which contributes to
improving the aerobic function of skeletal muscles, and ameliorat- long-term effects on gene expression.339 Forced circadian change
ing the loss of muscle fibers and turnover of motor neurons.322–325 may also accelerate the aging process and impair body function at
The effect of CR on lifespan and rejuvenation is at least partially a systemic level. Light schedule changes significantly affect aged
due to the amelioration of aging-related epigenetic changes, such mice, and advanced daytime leads to increased mortality of aged
as DNA methylation and histone modification.320,326 It is likely that mice.26 In addition, mice with an innate circadian period close to
epitranscriptomic regulation also functions as an effector of CR. For 24 h live 20% longer than those with a shorter or longer innate
example, CR in rats significantly inhibited the aging-associated circadian period.340 The disturbance of the circadian rhythm in
down-regulation of the RNA m6A reader protein YBX1, which has rodents indicates that maintaining regular day/night cycles may
been shown to be one of the drivers of stem cell aging.327 reduce aging-related mortality and raise the question of whether
According to the epigenetic clock developed in mice, 40% CR circadian rhythm affects humans. Notably, a short-term circadian
treatment slows the molecular changes and reduces the epigenetic misalignment of 12-h inverted behavioral and environmental
age in mouse livers. Notably, a 20-year CR, which reaches a final cycles for three days increases blood pressure and inflammatory
level at 30%, shows reduced aging-related pathologies and a markers in humans.341 However, how regular circadian rhythm
significant lifespan expansion in adult rhesus monkeys, indicating benefits the healthspan, especially from the lens of epigenetic
that moderate CR also exerts a rejuvenating effect on pri- mechanisms, still needs further investigation.
mates.328,329 In humans, moderate CR slows biological aging,
improves the function of the liver, and reduces oxidative stress and Exercise
the incidence of aging-related diseases.330–336 In summary, CR has Exercise may remodel DNA methylation on the promoter of key
been widely proven to be an effective method to delay aging, and genes in skeletal muscle342,343 and histone modifications could
clinical trials demonstrate the accessibility to applying CR in also be changed by exercise through inhibition of the function of

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HDACs, thereby influencing the gene expression patterns.344,345 Exercise has been demonstrated to be an effective geropro-
Moreover, exercise can modulate the expression of several tector to improve the lifespan and healthspan in humans.
miRNAs that mediate the beneficial process.346,347 After voluntary Vigorous exercise, such as running, at middle and older ages, is
resistance training for 8 weeks, aged mice exhibit nearly 8 weeks associated with reduced disability in later life and reduced
of younger epigenetic age in their muscle and a modest lifespan mortality,364,365 and leisure time physical activity of moderate to
extension.56 In addition, voluntary wheel running benefits aged vigorous intensity is associated with longer life expectancy.366 In
mice in neurogenesis and learning ability348 and reduces the clinical intervention studies, exercise is found to reverse a series of
abnormal changes of the aged synapse.324 Importantly, the aging-related diseases, including heart failure,367–371 cognitive
rejuvenating effect of exercise is also observed in humans. There decline,372,373 atherosclerosis,374 and insulin resistance.375,376 The
is a significant difference in the transcriptional profile between geroprotective effect of exercise in humans is closely linked to
physically active and sedentary aged adults, and endurance epigenetic regulation. Exercise modifies the DNA methylation
exercise improves the function of muscles in aged people.349 In patterns in aged human skeletal muscle and reduces stochastic
addition, resistance training reduces the level of the mitochondrial epigenetic mutations in crucial cancer-related pathways.32,363
methylome in aged human skeletal muscle and partially restores Endurance exercise upregulates the expression of SIRT3 in the
the aging-related change in the nuclear gene methylome in skeletal muscle and upregulates SIRT1, SIRT3, and SIRT6 in the
muscle.350,351 Clinical trials aiming to investigate the beneficial serum.375,377,378 Exercise also modulates the microRNA expression
effects of exercise will be discussed in the “Clinical intervention” profile (such as miR-423-3p, miR-451a, miR-766-3p, miR-130a, and
section later. miRNA-223) in subjects with type 2 diabetes, which may be
Current studies have demonstrated that a healthy lifestyle involved in the improvement of weight loss, blood glucose
indeed exerts a beneficial effect on aging and therefore raises control, and insulin sensitivity.379–381 In addition, the combination
awareness of vibrant health. However, caloric restriction, circadian of diet and lifestyle interventions, including exercise, sleep,
control and exercise all need to be moderate during implementa- relaxation guidance, supplemental probiotics and phytonutrients,
tion, and the boundary between healthy and unhealthy status reverses the epigenetic age in healthy adult males.362
requires further investigation. As our understanding of aging Pharmacological intervention is another major strategy to
deepens, a healthy lifestyle is considered to be the easiest way for target natural aging. Epigenetic-related compounds, such as
humans to interfere with aging, and active health definitively NMN, NR, and STACs, show potential as geroprotectors in clinical
deserves more attention. trials. Supplementation with NMN increases muscle insulin
sensitivity, insulin signaling, and muscle remodeling in prediabetic
Clinical intervention women.382 NMN prevents aging-related muscle dysfunctions383
Although various strategies targeting aging show satisfactory and shows benefits in improving aerobic capacity, cardiovascular
results in animal models, their effects in humans have yet to be fitness, sleep quality, fatigue, and physical performance.89,384 As
demonstrated. Currently, diet intervention and exercise, which are for NR, clinical trials indicate that NR suppresses inflammatory
extensively associated with epigenetic regulation, are the mostly activation of PBMCs in heart failure patients385 and decreases the
studied and accepted strategies to target aging in humans. The levels of inflammatory cytokines in the serum and cerebrospinal
clinical trial results demonstrate that CR (11.9%–25%) attenuates fluid of Parkinson’s disease (PD) patients.386 However, most of
aging-related biomarkers, such as decreasing weight, enhancing these studies focus on the safety and tolerability of NR in
insulin sensitivity and glucose tolerance, and improving major patients.386,387 The effectiveness of NR in preventing or attenuat-
cardiometabolic risk factors.334,352,353 Time-restricted eating (TRE) ing the progression of aging-related disorders should be verified
in humans also provides benefits to some extent. Under an 8–10 h in further studies, considering that several clinical trials show that
daily eating window of TRE, reductions in weight, blood pressure, NR does not improve insulin resistance.388–391 STACs exhibit
atherogenic lipids, and cardiovascular risks are observed.354,355 A inspiring effects in preclinical studies, but the results in clinical
more stringent TRE (6 h window) also shows improvement in trials are not as satisfactory. For example, the natural STAC
insulin sensitivity.356 However, the strategy of TRE is challenging to resveratrol and early synthetic STACs such as SRT1720 have very
undertake, especially for cases with longer fasting times. More- low bioavailability, potency, and limited target specificity,392 and
over, skipping breakfast has been found to be associated with an other STACs, such as SRT2379 and SRT3025, produce no significant
increased risk of mortality from cardiovascular disease.357 There- clinical responses. Similarly, although SRT2104 shows some
fore, an 11–12 h daily eating period is suggested to be ideal to benefits on lipid parameters, including cholesterol and triglycer-
avoid the compliance issues and side effects of TRE.358 Consider- ides,393,394 it does not improve glucose or insulin control394,395
ing the difficulty for most subjects to adhere to chronic and and has no significant anti-inflammatory effect in ulcerative colitis
extreme diets of CR or TRE, a fasting-mimicking diet (FMD) with patients.396 The poor and variable pharmacokinetics upon oral
low calories, low sugars, and low proteins but high unsaturated administration of SRT2104 need to be resolved in the future.
fats, provides another choice for a diet intervention. In a Other geroprotective interventions, such as metformin, rapa-
randomized phase 2 trial, healthy participants who received mycin, and D + Q, have also been explored in clinical trials, and
3 monthly 5-day FMD cycles exhibited reduced markers/risk the data show that metformin administration reduces the
factors for aging, diabetes, cancer, and cardiovascular disease.359 incidence of diabetes,397 cardiovascular events,398 frailty,399 and
In addition, a comprehensive understanding of the dietary cognitive impairment,400 and improves putative longevity effec-
interventions in humans has led to the proposal of the everyday tors in PBMCs.401 Although rapamycin shows exciting effects in
normocaloric longevity diet that includes a mid to high preclinical studies, similar results have not been observed in
carbohydrate and low but sufficient protein intake that is mostly clinical trials. In addition, despite the improved immune function
plant-based but includes regular consumption of pesco- in the elderly after administration of the mTOR inhibitor
vegetarian-derived proteins.358 Diet intervention has also been RAD001,402,403 several studies show that rapamycin does not
found to be associated with epigenetic regulation in clinical trials. improve cognitive function or physical performance404 and does
For example, CR in healthy and slightly overweight subjects not improve frailty.405 The first clinical study of senolytics
significantly increases plasma concentrations of SIRT1.360 Five days demonstrated that D + Q improves 6-min walk distance, walking
of periodic fasting significantly elevated the expression of SIRT1 speed, chair raise ability, and short physical performance battery
and SIRT3 in blood cells.361 Moreover, diet intervention has been in idiopathic pulmonary fibrosis (IPF) patients.309 D + Q also
shown to slow down the DNA methylation-based biomarkers of reduces senescent cell burden in adipose tissue and skin and
aging in several studies.362,363 reduces circulating SASP in people with diabetic kidney disease.406

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 Epigenetic regulation of aging: implications for interventions of aging. . .
Wang et al.
15
conditions; however, the safety concerns and inconsistent results
Table 1. Ongoing clinical trials related to epigenetic targets and
of these strategies demand further clinical evidence. Many other
regulation of aging
clinical trials related to epigenetic targets and the regulations of
Interventions Conditions/diseases Trial nO. Phase aging are still ongoing (Table 1). Together, these findings will
provide more candidates for gerotherapeutics and pave the way
Diet intervention Epigenetic aging NCT04962464 2 for fighting aging in the future.
Epigenetic aging NCT05297097 2
Epigenetic aging NCT05234203 NA
CONCLUSION AND PERSPECTIVE
Aging NCT05424042 NA
Studies in C. elegans, Drosophila, and mammals have unraveled
Exercise Aging NCT05232968 NA the aging-related epigenetic changes in DNA, RNA, and histone
Aging; Alzheimer disease NCT04299308 NA modifications and alterations in the more advanced chromatin
Biological aging NCT03440099 NA structure states. Correspondingly, these epigenetic changes have
Aging; Inflammatory NCT05042167 NA been identified as biomarkers or intervention targets of aging,
response such as the global decrease in genomic DNA methylation, the
global loss of canonical histones, chromatin landscape remodeling
NMN Glucose metabolism NCT04571008 NA
disorders caused by heterochromatin loss, and nuclear membrane protein
changes in human and mouse tissues during aging. However, the
Hypertension NCT04903210 4
same chromatin modifications (e.g., H3K14ac and H3K27me3) may
Physical activity; Muscle NCT04664361 NA play opposite roles in regulating aging and longevity across
recovery species and even across tissues within the same species,
NR Aging NCT03818802 NA indicating that epigenetic changes need to be interpreted with
Parkinson disease NCT03568968 NA their context. It is noteworthy that emerging technologies such as
Overweight and obesity; NCT04907110 NA single-cell omics sequencing provide a higher resolution for
Aging; Type 2 diabetes dissecting epigenetic characteristics during aging, and provide
Sarcopenia; Nicotinamide NCT04691986 NA new avenues for investigating the heterogeneity of aged cells. In
adenine dinucleotide addition, the spatiotemporal transcriptomic atlas across multiple
concentration; Muscle mammalian tissues can provide more information on aging-
quality and NAD+ content related interactions between cells or tissues, which may facilitate
STACs Vascular resistance; NCT01842399 1/2 the design of better and more precise therapeutics for aging and
Hypertension aging-related diseases.
Healthy NCT00996229 3 Based on these epigenetic changes in cells during aging, a
series of corresponding therapeutic strategies have been devel-
Metformin Epigenetic aging; NCT04375657 2
oped. Geroprotective drugs targeting longevity-related histone
Immunosenescence
acetylation, including supplementation with NAD+ precursors,
Aging; Insulin sensitivity; NCT04264897 3 STACs have been tested in various species. Metformin, rapamycin,
Chronic disease;
Mitochondria; Insulin
and other drugs have also shown positive effects in alleviating
resistance aging-related pathologies and regulating aging-related epigenetic
changes in preclinical studies; however, the safety and efficacy of
Frailty; Sarcopenic NCT04221750 3
obesity; Aging
these drugs require more clinical investigation. Currently, a
rational diet and exercise are considered the most effective and
Frailty NCT02570672 2 easiest way to delay aging, but drugs targeting key aging-related
Mild cognitive impairment NCT04098666 2/3 molecular and cellular changes are still promising and attractive
Insulin resistance; Obesity NCT03733132 2 clinical treatment strategies for intervening in aging and treating
Rapamycin Epigenetic clock of skin NCT04608448 1 aging-related diseases.
Aging NCT04488601 2
Despite all the recent progress, it remains unclear how
epigenetic changes interact with other factors, including the
Aging NCT04742777 2 genetic background and even the microbiome, to regulate the
Mild cognitive impairment; NCT04629495 2 aging process. It is also unclear how environmental factors,
Alzheimer disease lifestyles, and physiological and psychological states contribute to
Mild cognitive impairment; NCT04200911 1 epigenetic changes in the aging process. Furthermore, as aging is
Alzheimer disease a continuous process that occurs over many years in humans, it
D+Q Epigenetic aging NCT04946383 2 would be necessary to track the epigenetic changes in this entire
Diabetic kidney disease NCT02848131 1 process for a better understanding of what and how epigenetic
regulations contribute to each stage of aging.
Alzheimer disease NCT04063124 1/2
Mild cognitive impairment; NCT04785300 1/2
Alzheimer disease
ACKNOWLEDGEMENTS
Alzheimer disease; Mild NCT04685590 2 The authors apologize for not citing all important studies in this review due to
cognitive impairment constraints on manuscript length. This work was supported by the National Key
Aging-related osteoporosis NCT04313634 2 Research and Development Program of China (2020YFA0804000), the Strategic
Priority Research Program of the Chinese Academy of Sciences (XDA16010000), CAS
Project for Young Scientists in Basic Research (YSBR-076, YSBR-012), the National
Natural Science Foundation of China (81921006, 92149301, 92168201, 92049116,
Collectively, diet intervention and exercise are still the most
32121001, 82100140, 31970597), the National Key Research and Development
accepted strategies to intervene in aging and aging-related Program of China (2020YFA0803401 2019YFA0802202), the Program of the Beijing
diseases, mainly because of their effectiveness and safety for Natural Science Foundation (Z190019), the Tencent Foundation (2021–1045), K. C.
humans. Advances in pharmacological interventions such as Wong Education Foundation (GJTD-2019-08), Youth Innovation Promotion Associa-
geroprotectors also show improvement in multiple aging-related tion of CAS (E1CAZW0401), Science & Technology Innovation 2030 of The Ministry of

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 Epigenetic regulation of aging: implications for interventions of aging. . .
Wang et al.
16
Science and Technology of China (2022ZD0214200). All figures were created with 28. Liu, G. H. et al. Recapitulation of premature ageing with iPSCs from Hutchinson-
Biorender.com. Gilford progeria syndrome. Nature 472, 221–225 (2011).
29. Zhu, Y. et al. The Achilles’ heel of senescent cells: from transcriptome to
senolytic drugs. Aging Cell 14, 644–658 (2015).
AUTHOR CONTRIBUTIONS 30. Suda, M. et al. Senolytic vaccination improves normal and pathological age-
G.-H.L., F.Z., J.R., and W.Z. designed and supervised the review, and reviewed the related phenotypes and increases lifespan in progeroid mice. Nat. Aging 1,
manuscript; K.W., H.L., Q.H., L.W., J.L. wrote the manuscript and drew pictures and 1117–1126 (2021).
tables; Z.Z. performed manuscript reviewing and editing; all authors have read and 31. Johnson, A. A. et al. The role of DNA methylation in aging, rejuvenation, and
approved the article. age-related disease. Rejuvenation Res. 15, 483–494 (2012).
32. Sailani, M. R. et al. Lifelong physical activity is associated with promoter hypo-
methylation of genes involved in metabolism, myogenesis, contractile proper-
ties and oxidative stress resistance in aged human skeletal muscle. Sci. Rep. 9,
ADDITIONAL INFORMATION
3272 (2019).
Competing interests: The authors declare no competing interests. 33. Maegawa, S. et al. Widespread and tissue specific age-related DNA methylation
changes in mice. Genome Res. 20, 332–340 (2010).
34. Adhikari, S. & Curtis, P. D. DNA methyltransferases and epigenetic regulation in
REFERENCES bacteria. FEMS Microbiol. Rev. 40, 575–591 (2016).
1. Catana, C. S., Atanasov, A. G. & Berindan-Neagoe, I. Natural products with anti- 35. Li, Y. et al. Stella safeguards the oocyte methylome by preventing de novo
aging potential: affected targets and molecular mechanisms. Biotechnol. Adv. 36, methylation mediated by DNMT1. Nature 564, 136–140 (2018).
1649–1656 (2018). 36. Baets, J. et al. Defects of mutant DNMT1 are linked to a spectrum of neurological
2. Brunet, A. & Berger, S. L. Epigenetics of aging and aging-related disease. J. disorders. Brain 138, 845–861 (2015).
Gerontol. A Biol. Sci. Med. Sci. 69, S17–S20 (2014). 37. Zhang, Z.-M. et al. Structural basis for DNMT3A-mediated de novo DNA
3. Lopez-Otin, C. et al. The hallmarks of aging. Cell 153, 1194–1217 (2013). methylation. Nature 554, 387–391 (2018).
4. Anderson, K. E. et al. The queen’s gut refines with age: longevity phenotypes in a 38. Yagi, M. et al. Identification of distinct loci for de novo DNA methylation by
social insect model. Microbiome 6, 108 (2018). DNMT3A and DNMT3B during mammalian development. Nat. Commun. 11,
5. Horvath, S. et al. DNA methylation clocks tick in naked mole rats but queens age 3199 (2020).
more slowly than nonbreeders. Nat. Aging 2, 46–59 (2022). 39. Verma, N. et al. TET proteins safeguard bivalent promoters from de novo
6. Tomczyk, S., Fischer, K., Austad, S., & Galliot, B. Hydra, a powerful model for methylation in human embryonic stem cells. Nat. Genet. 50, 83–95 (2018).
aging studies. Invertebr. Reprod. Dev. 59, 11–16 (2015). 40. Bick, A. G. et al. Genetic interleukin 6 signaling deficiency attenuates cardio-
7. Montesanto, A. et al. Epidemiological, genetic and epigenetic aspects of the vascular risk in clonal hematopoiesis. Circulation 141, 124–131 (2020).
research on healthy ageing and longevity. Immun. Ageing 9, 6 (2012). 41. Heyn, H. et al. Distinct DNA methylomes of newborns and centenarians. Proc.
8. Kubben, N. & Misteli, T. Shared molecular and cellular mechanisms of premature Natl Acad. Sci. USA 109, 10522–10527 (2012).
ageing and ageing-associated diseases. Nat. Rev. Mol. Cell Biol. 18, 595–609 (2017). 42. Zhang, J. et al. Highly enriched BEND3 prevents the premature activation of
9. Sun, Y., Li, Q. & Kirkland, J. L. Targeting senescent cells for a healthier longevity: bivalent genes during differentiation. Science 375, 1053–1058 (2022).
the roadmap for an era of global aging. Life Med. lnac030 https://doi.org/ 43. Seale, K. et al. Making sense of the ageing methylome. Nat. Rev. Genet. 23,
10.1002/advs.202002611 (2022). 585–605 (2022).
10. Cai, Y. et al. The landscape of aging. Sci. China Life Sci. 1–101 https://doi.org/ 44. Ahuja, N. et al. Aging and DNA methylation in colorectal mucosa and cancer.
10.1007/s11427-022-2161-3 (2022). Cancer Res. 58, 5489–5494 (1998).
11. Berdyshev, G. D., Korotaev, G. K., Boiarskikh, G. V. & Vaniushin, B. F. [Nucleotide 45. Waki, T., Tamura, G., Sato, M. & Motoyama, T. Age-related methylation of tumor
composition of DNA and RNA from somatic tissues of humpback and its suppressor and tumor-related genes: an analysis of autopsy samples. Oncogene
changes during spawning]. Biokhimiia 32, 988–993 (1967). 22, 4128–4133 (2003).
12. Wilson, V. L. & Jones, P. A. DNA methylation decreases in aging but not in 46. Kim, J. Y., Siegmund, K. D., Tavare, S. & Shibata, D. Age-related human small
immortal cells. Science 220, 1055–1057 (1983). intestine methylation: evidence for stem cell niches. BMC Med. 3, 10 (2005).
13. Wilson, V. L., Smith, R. A., Ma, S. & Cutler, R. G. Genomic 5-methyldeoxycytidine 47. Lu, A. T. et al. Universal DNA methylation age across mammalian tissues. Pre-
decreases with age. J. Biol. Chem. 262, 9948–9951 (1987). print at bioRxiv https://doi.org/10.1101/2021.01.18.426733 (2021).
14. Ishimi, Y. et al. Changes in chromatin structure during aging of human skin 48. Talens, R. P. et al. Epigenetic variation during the adult lifespan: cross-sectional
fibroblasts. Exp. Cell Res. 169, 458–467 (1987). and longitudinal data on monozygotic twin pairs. Aging Cell 11, 694–703 (2012).
15. Greer, E. L. et al. Members of the H3K4 trimethylation complex regulate lifespan 49. Slieker, R. C. et al. Age-related accrual of methylomic variability is linked to
in a germline-dependent manner in C. elegans. Nature 466, 383–387 (2010). fundamental ageing mechanisms. Genome Biol. 17, 191 (2016).
16. Horvath, S. DNA methylation age of human tissues and cell types. Genome Biol. 50. Lowe, R. et al. Ageing-associated DNA methylation dynamics are a molecular
14, R115 (2013). readout of lifespan variation among mammalian species. Genome Biol. 19, 22 (2018).
17. Li, Q. et al. NSUN2-mediated m5C methylation and METTL3/METTL14-mediated 51. Bocklandt, S. et al. Epigenetic predictor of age. PLoS ONE 6, e14821 (2011).
m6A methylation cooperatively enhance p21 translation. J. Cell Biochem. 118, 52. Hannum, G. et al. Genome-wide methylation profiles reveal quantitative views
2587–2598 (2017). of human aging rates. Mol. Cell 49, 359–367 (2013).
18. Cheung, P. et al. Single-cell chromatin modification profiling reveals increased 53. Levine, M. E. et al. An epigenetic biomarker of aging for lifespan and healthspan.
epigenetic variations with aging. Cell 173, 1385–1397 e1314 (2018). Aging 10, 573–591 (2018).
19. Zou, X. et al. From monkey single-cell atlases into a broader biomedical per- 54. Horvath, S. & Raj, K. DNA methylation-based biomarkers and the epigenetic
spective. Life Med. lnac028 https://doi.org/10.1093/lifemedi/lnac028 (2022). clock theory of ageing. Nat. Rev. Genet. 19, 371–384 (2018).
20. Aging Atlas, C. Aging Atlas: a multi-omics database for aging biology. Nucleic 55. McCrory, C. et al. GrimAge outperforms other epigenetic clocks in the prediction
Acids Res. 49, D825–D830 (2021). of age-related clinical phenotypes and all-cause mortality. J. Gerontol. A Biol. Sci.
21. McCay, C. M., Crowell, M. F. & Maynard, L. A. The effect of retarded growth upon Med. Sci. 76, 741–749 (2021).
the length of life span and upon the ultimate body size: one figure. J. Nutr. 10, 56. Murach, K. A. et al. Late-life exercise mitigates skeletal muscle epigenetic aging.
63–79 (1935). Aging Cell 21, e13527 (2022).
22. Acosta-Rodriguez, V. et al. Circadian alignment of early onset caloric restriction 57. Trapp, A., Kerepesi, C. & Gladyshev, V. N. Profiling epigenetic age in single cells.
promotes longevity in male C57BL/6J mice. Science 376, 1192–1202 (2022). Nat. Aging 1, 1189–1201 (2021).
23. Kennedy, B. K., Austriaco, N. R. Jr, Zhang, J. & Guarente, L. Mutation in the 58. Kouzarides, T. Chromatin modifications and their function. Cell 128, 693–705
silencing gene SIR4 can delay aging in S. cerevisiae. Cell 80, 485–496 (1995). (2007).
24. Howitz, K. T. et al. Small molecule activators of sirtuins extend Saccharomyces 59. Sidler, C., Kovalchuk, O. & Kovalchuk, I. Epigenetic regulation of cellular senes-
cerevisiae lifespan. Nature 425, 191–196 (2003). cence and aging. Front. Genet. 8, 138 (2017).
25. Conboy, I. M. et al. Rejuvenation of aged progenitor cells by exposure to a 60. Santos-Rosa, H. et al. Active genes are tri-methylated at K4 of histone H3. Nature
young systemic environment. Nature 433, 760–764 (2005). 419, 407–411 (2002).
26. Davidson, A. J. et al. Chronic jet-lag increases mortality in aged mice. Curr. Biol. 61. Bernstein, B. E. et al. Methylation of histone H3 Lys 4 in coding regions of active
16, R914–R916 (2006). genes. Proc. Natl Acad. Sci. USA 99, 8695–8700 (2002).
27. Lapasset, L. et al. Rejuvenating senescent and centenarian human cells by 62. Cruz, C. et al. Tri-methylation of histone H3 lysine 4 facilitates gene expression in
reprogramming through the pluripotent state. Genes Dev. 25, 2248–2253 (2011). ageing cells. Elife. 7, https://doi.org/10.7554/elife.34081 (2018).

Signal Transduction and Targeted Therapy (2022)7:374

 Epigenetic regulation of aging: implications for interventions of aging. . .
Wang et al.
17
63. Ren, X. et al. Maintenance of nucleolar homeostasis by CBX4 alleviates senes- 94. Huang, B. et al. Inhibition of histone acetyltransferase GCN5 extends lifespan in
cence and osteoarthritis. Cell Rep. 26, 3643–3656 e3647 (2019). both yeast and human cell lines. Aging Cell 19, e13129 (2020).
64. Pu, M. et al. Unique patterns of trimethylation of histone H3 lysine 4 are prone 95. Di Giorgio, E. et al. HDAC4 degradation during senescence unleashes an epi-
to changes during aging in Caenorhabditis elegans somatic cells. PLoS Genet. genetic program driven by AP-1/p300 at selected enhancers and super-
14, e1007466 (2018). enhancers. Genome Biol. 22, 129 (2021).
65. Bazopoulou, D. et al. Developmental ROS individualizes organismal stress 96. Sen, P. et al. Histone acetyltransferase p300 induces de novo super-enhancers to
resistance and lifespan. Nature 576, 301–305 (2019). drive cellular senescence. Mol. Cell 73, 684–698 e688 (2019).
66. Cao, Q. et al. Targeting histone K4 trimethylation for treatment of cognitive and 97. Barrett, L. N. & Westerheide, S. D. The CBP-1/p300 Lysine Acetyltransferase
synaptic deficits in mouse models of Alzheimer’s disease. Sci. Adv. 6, eabc8096 Regulates the Heat Shock Response in C. elegans. Front. Aging. 3, 861761 (2022).
(2020). 98. Li, T. Y. et al. The transcriptional coactivator CBP/p300 is an evolutionarily
67. Sun, D. et al. Epigenomic profiling of young and aged HSCs reveals concerted conserved node that promotes longevity in response to mitochondrial stress.
changes during aging that reinforce self-renewal. Cell Stem Cell 14, 673–688 (2014). Nat. Aging 1, 165–178 (2021).
68. Adelman, E. R. et al. Aging human hematopoietic stem cells manifest profound 99. Tasdemir, N. et al. BRD4 connects enhancer remodeling to senescence immune
epigenetic reprogramming of enhancers that may predispose to leukemia. surveillance. Cancer Discov. 6, 612–629 (2016).
Cancer Discov. 9, 1080–1101 (2019). 100. Marzi, S. J. et al. A histone acetylome-wide association study of Alzheimer’s
69. Cheung, I. et al. Developmental regulation and individual differences of neu- disease identifies disease-associated H3K27ac differences in the entorhinal
ronal H3K4me3 epigenomes in the prefrontal cortex. Proc. Natl Acad. Sci. USA cortex. Nat. Neurosci. 21, 1618–1627 (2018).
107, 8824–8829 (2010). 101. Klein, H. U. et al. Epigenome-wide study uncovers large-scale changes in histone
70. Bonasio, R., Tu, S. & Reinberg, D. Molecular signals of epigenetic states. Science acetylation driven by tau pathology in aging and Alzheimer’s human brains.
330, 612–616 (2010). Nat. Neurosci. 22, 37–46 (2019).
71. Shumaker, D. K. et al. Mutant nuclear lamin A leads to progressive alterations of 102. Price, A. J. et al. Hdac3, Setdb1, and Kap1 mark H3K9me3/H3K14ac bivalent
epigenetic control in premature aging. Proc. Natl Acad. Sci. USA 103, 8703–8708 regions in young and aged liver. Aging Cell. 19, e13092 (2020).
(2006). 103. Singh, P. & Thakur, M. K. Histone deacetylase 2 inhibition attenuates down-
72. Baumgart, M. et al. RNA-seq of the aging brain in the short-lived fish N. furzeri - regulation of hippocampal plasticity gene expression during aging. Mol. Neu-
conserved pathways and novel genes associated with neurogenesis. Aging Cell robiol. 55, 2432–2442 (2018).
13, 965–974 (2014). 104. Wang, W. et al. A genome-wide CRISPR-based screen identifies KAT7 as a driver
73. Jin, C. et al. Histone demethylase UTX-1 regulates C. elegans life span by tar- of cellular senescence. Sci. Transl. Med. 13, eabd2655 (2021).
geting the insulin/IGF-1 signaling pathway. Cell Metab. 14, 161–172 (2011). 105. Tasselli, L. et al. SIRT6 deacetylates H3K18ac at pericentric chromatin to prevent
74. Guillermo, A. R. R. et al. H3K27 modifiers regulate lifespan in C. elegans in a mitotic errors and cellular senescence. Nat. Struct. Mol. Biol. 23, 434–440 (2016).
context-dependent manner. BMC Biol. 19, 59 (2021). 106. Feser, J. et al. Elevated histone expression promotes life span extension. Mol. Cell
75. Maures, T. J., Greer, E. L., Hauswirth, A. G. & Brunet, A. The H3K27 demethylase 39, 724–735 (2010).
UTX-1 regulates C. elegans lifespan in a germline-independent, insulin- 107. Hachinohe, M., Hanaoka, F. & Masumoto, H. Hst3 and Hst4 histone deacetylases
dependent manner. Aging Cell 10, 980–990 (2011). regulate replicative lifespan by preventing genome instability in Saccharomyces
76. Merkwirth, C. et al. Two conserved histone demethylases regulate mitochondrial cerevisiae. Genes Cells 16, 467–477 (2011).
stress-induced longevity. Cell 165, 1209–1223 (2016). 108. Dang, W. et al. Histone H4 lysine 16 acetylation regulates cellular lifespan.
77. Sen, P. et al. H3K36 methylation promotes longevity by enhancing transcrip- Nature 459, 802–807 (2009).
tional fidelity. Genes Dev. 29, 1362–1376 (2015). 109. Nativio, R. et al. Dysregulation of the epigenetic landscape of normal aging in
78. Jeon, H. J. et al. Effect of heterochromatin stability on intestinal stem cell aging Alzheimer’s disease. Nat. Neurosci. 21, 497–505 (2018).
in Drosophila. Mech. Ageing Dev. 173, 50–60 (2018). 110. Peleg, S. et al. Altered histone acetylation is associated with age-dependent
79. Li, C. L. et al. Region-specific H3K9me3 gain in aged somatic tissues in Cae- memory impairment in mice. Science 328, 753–756 (2010).
norhabditis elegans. PLoS Genet. 17, e1009432 (2021). 111. Benito, E. et al. HDAC inhibitor-dependent transcriptome and memory rein-
80. Wood, J. G. et al. Chromatin remodeling in the aging genome of Drosophila. statement in cognitive decline models. J. Clin. Invest. 125, 3572–3584 (2015).
Aging Cell 9, 971–978 (2010). 112. Yang, L. et al. Ubiquitylome study identifies increased histone 2A ubiquitylation
81. Zhang, W. et al. Aging stem cells. A Werner syndrome stem cell model unveils as an evolutionarily conserved aging biomarker. Nat. Commun. 10, 2191 (2019).
heterochromatin alterations as a driver of human aging. Science 348, 1160–1163 113. Joos, J. P. et al. Ectopic expression of S28A-mutated Histone H3 modulates
(2015). longevity, stress resistance and cardiac function in Drosophila. Sci. Rep. 8, 2940
82. Wu, Z. et al. Differential stem cell aging kinetics in Hutchinson-Gilford progeria (2018).
syndrome and Werner syndrome. Protein Cell 9, 333–350 (2018). 114. Yung, P. Y. K. et al. Histone H3 serine 28 is essential for efficient polycomb-
83. Liu, Z. et al. Large-scale chromatin reorganization reactivates placenta-specific mediated gene repression in Drosophila. Cell Rep. 11, 1437–1445 (2015).
genes that drive cellular aging. Dev. Cell 57, 1347–1368 e1312 (2022). 115. Luger, K. et al. Crystal structure of the nucleosome core particle at 2.8 A reso-
84. Djeghloul, D. et al. Age-associated decrease of the histone methyltransferase lution. Nature 389, 251–260 (1997).
SUV39H1 in HSC perturbs heterochromatin and B lymphoid differentiation. Stem 116. Chandra, T. et al. Global reorganization of the nuclear landscape in senescent
Cell Rep. 6, 970–984 (2016). cells. Cell Rep. 10, 471–483 (2015).
85. Li, Y. et al. Vitamin C alleviates aging defects in a stem cell model for Werner 117. Zhao, D. & Chen, S. Failures at every level: breakdown of the epigenetic
syndrome. Protein Cell 7, 478–488 (2016). machinery of aging. Life Medicine, lnac016, https://doi.org/10.1093/lifemedi/
86. Shan, H. et al. Large-scale chemical screen identifies Gallic acid as a ger- lnac016 (2022).
oprotector for human stem cells. Protein Cell 13, 532–539 (2021). 118. Hu, Z. et al. Nucleosome loss leads to global transcriptional up-regulation and
87. Li, W. et al. Low-dose chloroquine treatment extends the lifespan of aged rats. genomic instability during yeast aging. Genes Dev. 28, 396–408 (2014).
Protein Cell 13, 454–461 (2022). 119. O’Sullivan, R. J., Kubicek, S., Schreiber, S. L. & Karlseder, J. Reduced histone
88. Zhang, Y. et al. Single-nucleus transcriptomics reveals a gatekeeper role for biosynthesis and chromatin changes arising from a damage signal at telomeres.
FOXP1 in primate cardiac aging. Protein Cell pwac038, https://doi.org/10.1093/ Nat. Struct. Mol. Biol. 17, 1218–1225 (2010).
procel/pwac038 (2022). 120. Chen, Y. et al. Remodeling of the H3 nucleosomal landscape during mouse
89. Liao, B. et al. Nicotinamide mononucleotide supplementation enhances aerobic aging. Transl. Med. Aging 4, 22–31 (2020).
capacity in amateur runners: a randomized, double-blind study. J. Int. Soc. Sports 121. Flex, E. et al. Aberrant function of the C-terminal tail of HIST1H1E accelerates
Nutr. 18, 54 (2021). cellular senescence and causes premature aging. Am. J. Hum. Genet. 105,
90. Mostoslavsky, R. et al. Genomic instability and aging-like phenotype in the 493–508 (2019).
absence of mammalian SIRT6. Cell 124, 315–329 (2006). 122. Liu, B., Yip, R. & Zhou, Z. Chromatin remodeling, DNA damage repair and aging.
91. Tanner, K. G., Landry, J., Sternglanz, R. & Denu, J. M. Silent information regulator 2 Curr. Genomics 13, 533–547 (2012).
family of NAD- dependent histone/protein deacetylases generates a unique pro- 123. Zhu, D. et al. NuRD mediates mitochondrial stress-induced longevity via chro-
duct, 1-O-acetyl-ADP-ribose. Proc. Natl Acad. Sci. USA 97, 14178–14182 (2000). matin remodeling in response to acetyl-CoA level. Sci. Adv. 6, eabb2529 (2020).
92. Gong, H. et al. Age-dependent tissue expression patterns of Sirt1 in senescence- 124. Zhu, D., Li, X. & Tian, Y. Mitochondrial-to-nuclear communication in aging: an
accelerated mice. Mol. Med. Rep. 10, 3296–3302 (2014). epigenetic perspective. Trends Biochem. Sci. 47, 645–659 (2022).
93. Cho, S. H. et al. SIRT1 deficiency in microglia contributes to cognitive decline in 125. Pouikli, A. et al. Chromatin remodeling due to degradation of citrate carrier
aging and neurodegeneration via epigenetic regulation of IL-1beta. J. Neurosci. impairs osteogenesis of aged mesenchymal stem cells. Nat. Aging 1, 810–825
35, 807–818 (2015). (2021).

Signal Transduction and Targeted Therapy (2022)7:374

 Epigenetic regulation of aging: implications for interventions of aging. . .
Wang et al.
18
126. Kind, J. et al. Single-cell dynamics of genome-nuclear lamina interactions. Cell 160. Lewinska, A., Adamczyk-Grochala, J., Kwasniewicz, E. & Wnuk, M. Down-
153, 178–192 (2013). regulation of methyltransferase Dnmt2 results in condition-dependent telomere
127. Stephens, A. D. et al. Chromatin histone modifications and rigidity affect nuclear shortening and senescence or apoptosis in mouse fibroblasts. J. Cell Physiol.
morphology independent of lamins. Mol. Biol. Cell 29, 220–233 (2018). 232, 3714–3726 (2017).
128. Fang, Y. et al. Loss of Atg7 causes chaotic nucleosome assembly of mouse bone 161. Lewinska, A., Adamczyk-Grochala, J., Deregowska, A. & Wnuk, M. Sulforaphane-
marrow CD11b(+)Ly6G(-) myeloid cells. Aging (Albany NY) 12, 25673–25683 induced cell cycle arrest and senescence are accompanied by DNA hypo-
(2020). methylation and changes in microRNA profile in breast cancer cells. Theranostics
129. Yu, R. et al. Cellular response to moderate chromatin architectural defects 7, 3461–3477 (2017).
promotes longevity. Sci. Adv. 5, eaav1165 (2019). 162. Sun, X. et al. Decreased expression of m(6)A demethylase FTO in ovarian aging.
130. Sun, L., Yu, R. & Dang, W. Chromatin architectural changes during cellular Arch. Gynecol. Obstet. 303, 1363–1369 (2021).
senescence and aging. Genes (Basel). 9, 211(2018). 163. Hirayama, M. et al. FTO demethylates cyclin D1 mRNA and controls cell-cycle
131. Wu, S. et al. BRM-SWI/SNF chromatin remodeling complex enables functional progression. Cell Rep. 31, 107464 (2020).
telomeres by promoting co-expression of TRF2 and TRF1. PLoS Genet. 16, 164. Li, G. et al. m6A hypomethylation of DNMT3B regulated by ALKBH5 promotes
e1008799 (2020). intervertebral disc degeneration via E4F1 deficiency. Clin. Transl. Med. 12, e765
132. Riedel, C. G. et al. DAF-16 employs the chromatin remodeller SWI/SNF to pro- (2022).
mote stress resistance and longevity. Nat. Cell Biol. 15, 491–501 (2013). 165. Zaccara, S., Ries, R. J. & Jaffrey, S. R. Reading, writing and erasing mRNA
133. Hoffmann, A. & Spengler, D. Chromatin remodeling complex NuRD in neuro- methylation. Nat. Rev. Mol. Cell Biol. 20, 608–624 (2019).
development and neurodevelopmental disorders. Front. Genet. 10, 682 (2019). 166. Chelmicki, T. et al. m(6)A RNA methylation regulates the fate of endogenous
134. Li, X. et al. Chromatin remodeling factor BAZ1A regulates cellular senescence in retroviruses. Nature 591, 312–316 (2021).
both cancer and normal cells. Life Sci. 229, 225–232 (2019). 167. Tam, O. H., Ostrow, L. W. & Gale Hammell, M. Diseases of the nERVous system:
135. Sun, L. et al. Attenuation of epigenetic regulator SMARCA4 and ERK-ETS sig- retrotransposon activity in neurodegenerative disease. Mob. DNA 10, 32 (2019).
naling suppresses aging-related dopaminergic degeneration. Aging Cell 19, 168. Fan, T. et al. Senolytics cocktail dasatinib and quercetin alleviate human
e13210 (2020). umbilical vein endothelial cell senescence via the TRAF6-MAPK-NF-kappaB Axis
136. Zhang, C. et al. ATF3 drives senescence by reconstructing accessible chromatin in a YTHDF2-dependent manner. Gerontology 68, 920–934 (2022).
profiles. Aging Cell 20, e13315 (2021). 169. Han, M. et al. Abnormality of m6A mRNA methylation is involved in Alzheimer’s
137. Virk, R. K. A. et al. Disordered chromatin packing regulates phenotypic plasticity. disease. Front. Neurosci. 14, 98 (2020).
Sci. Adv. 6, eaax6232 (2020). 170. Shafik, A. M. et al. N6-methyladenosine dynamics in neurodevelopment and
138. Lieberman-Aiden, E. et al. Comprehensive mapping of long-range interactions aging, and its potential role in Alzheimer’s disease. Genome Biol. 22, 17 (2021).
reveals folding principles of the human genome. Science 326, 289–293 (2009). 171. Xing, J. et al. NSun2 promotes cell growth via elevating cyclin-dependent kinase
139. Diao, Z. et al. SIRT3 consolidates heterochromatin and counteracts senescence. 1 translation. Mol. Cell Biol. 35, 4043–4052 (2015).
Nucleic Acids Res. 49, 4203–4219 (2021). 172. Tang, H. et al. NSun2 delays replicative senescence by repressing p27 (KIP1)
140. Bi, S. et al. SIRT7 antagonizes human stem cell aging as a heterochromatin translation and elevating CDK1 translation. Aging (Albany NY) 7, 1143–1158
stabilizer. Protein Cell 11, 483–504 (2020). (2015).
141. Hu, H. et al. ZKSCAN3 counteracts cellular senescence by stabilizing hetero- 173. Zhang, X. et al. The tRNA methyltransferase NSun2 stabilizes p16INK(4) mRNA
chromatin. Nucleic Acids Res. 48, 6001–6018 (2020). by methylating the 3’-untranslated region of p16. Nat. Commun. 3, 712 (2012).
142. Liang, C. et al. Stabilization of heterochromatin by CLOCK promotes stem cell 174. Cai, X. et al. RNA methyltransferase NSUN2 promotes stress-induced HUVEC
rejuvenation and cartilage regeneration. Cell Res. 31, 187–205 (2021). senescence. Oncotarget 7, 19099–19110 (2016).
143. Liu, X. et al. Resurrection of human endogenous retroviruses during aging 175. Tang, H. et al. HuR regulates telomerase activity through TERC methylation. Nat.
reinforces senescence. Preprint at bioRxiv https://doi.org/10.1101/ Commun. 9, 2213 (2018).
2021.02.22.432260(2021). 176. Slotkin, W. & Nishikura, K. Adenosine-to-inosine RNA editing and human disease.
144. Zhao, H. et al. Destabilizing heterochromatin by APOE mediates senescence. Genome Med. 5, 105 (2013).
Nat. Aging 2, 303–316 (2022). 177. Zhang, Y. et al. Comparative functional RNA editomes of neural differentiation from
145. Liang, C. et al. BMAL1 moonlighting as a gatekeeper for LINE1 repression and human PSCs. Life Med. lnac027, https://doi.org/10.1093/lifemedi/lnac027 (2022).
cellular senescence in primates. Nucleic Acids Res. 50, 3323–3347 (2022). 178. Gaisler-Salomon, I. et al. Hippocampus-specific deficiency in RNA editing of
146. Zhang, X. et al. The loss of heterochromatin is associated with multiscale three- GluA2 in Alzheimer’s disease. Neurobiol. Aging 35, 1785–1791 (2014).
dimensional genome reorganization and aberrant transcription during cellular 179. Stellos, K. et al. Adenosine-to-inosine RNA editing controls cathepsin S expres-
senescence. Genome Res. 31, 1121–1135 (2021). sion in atherosclerosis by enabling HuR-mediated post-transcriptional regula-
147. Zhang, W., Qu, J., Liu, G. H. & Belmonte, J. C. I. The ageing epigenome and its tion. Nat. Med. 22, 1140–1150 (2016).
rejuvenation. Nat. Rev. Mol. Cell Biol. 21, 137–150 (2020). 180. ElSharawy, A. et al. Genome-wide miRNA signatures of human longevity. Aging
148. Jin, F. et al. A high-resolution map of the three-dimensional chromatin inter- Cell 11, 607–616 (2012).
actome in human cells. Nature 503, 290–294 (2013). 181. Leng, S. X. & Pawelec, G. Single-cell immune atlas for human aging and frailty.
149. Ong, C. T. & Corces, V. G. CTCF: an architectural protein bridging genome Life Med. lnac013, https://doi.org/10.1093/lifemedi/lnac013 (2022).
topology and function. Nat. Rev. Genet. 15, 234–246 (2014). 182. Lim, Y. W. et al. Genome-wide DNA hypomethylation and RNA:DNA hybrid
150. Hou, Y. et al. CTCF mediates replicative senescence through POLD1. Front. Cell accumulation in Aicardi-Goutières syndrome. Elife. 4, e08007 (2015).
Dev. Biol. 9, 618586 (2021). 183. Crossley, M. P., Bocek, M. & Cimprich, K. A. R-loops as cellular regulators and
151. Hirosue, A. et al. Quantitative assessment of higher-order chromatin structure of genomic threats. Mol. Cell 73, 398–411 (2019).
the INK4/ARF locus in human senescent cells. Aging Cell 11, 553–556 (2012). 184. Kristensen, L. S. et al. The biogenesis, biology and characterization of circular
152. Frye, M., Harada, B. T., Behm, M. & He, C. RNA modifications modulate gene RNAs. Nat. Rev. Genet. 20, 675–691 (2019).
expression during development. Science 361, 1346–1349 (2018). 185. Wang, S. et al. Corrigendum to: Computational annotation of miRNA tran-
153. Sun, J. et al. The potential role of m6A RNA methylation in the aging process scription start sites. Brief. Bioinform. 22, 609 (2021).
and aging-associated diseases. Front. Genet. 13, 869950 (2022). 186. Wang, S. et al. Computational annotation of miRNA transcription start sites. Brief.
154. Oerum, S., Meynier, V., Catala, M. & Tisne, C. A comprehensive review of m6A/ Bioinform. 22, 380–392 (2021).
m6Am RNA methyltransferase structures. Nucleic Acids Res. 49, 7239–7255 (2021). 187. Wang, T. et al. Epigenetic aging signatures in mice livers are slowed by dwarfism,
155. Chen, X. et al. METTL3-mediated m(6)A modification of ATG7 regulates calorie restriction and rapamycin treatment. Genome Biol. 18, 57 (2017).
autophagy-GATA4 axis to promote cellular senescence and osteoarthritis pro- 188. Rivetti di Val Cervo, P. et al. p63-microRNA feedback in keratinocyte senescence.
gression. Ann. Rheum. Dis. 81, 87–99 (2022). Proc. Natl Acad. Sci. USA 109, 1133–1138 (2012).
156. Zhu, H. et al. N6-methyladenosine induced miR-34a-5p promotes TNF-alpha- 189. Xiang, X. et al. Cellular senescence in hepatocellular carcinoma induced by a
induced nucleus pulposus cell senescence by targeting SIRT1. Front. Cell Dev. long non-coding RNA-encoded peptide PINT87aa by blocking FOXM1-
Biol. 9, 642437 (2021). mediated. Theranostics 11, 4929–4944 (2021).
157. Li, G. et al. WTAP-mediated m(6)A modification of lncRNA NORAD promotes 190. Zhang, H. et al. LncRNA NEAT1 controls the lineage fates of BMSCs during
intervertebral disc degeneration. Nat. Commun. 13, 1469 (2022). skeletal aging by impairing mitochondrial function and pluripotency main-
158. Zhang, J. et al. Lamin A safeguards the m(6) A methylase METTL14 nuclear tenance. Cell Death Differ. 29, 351–365 (2022).
speckle reservoir to prevent cellular senescence. Aging Cell 19, e13215 (2020). 191. Yu, F. et al. Long non-coding RNA APTR promotes the activation of hepatic
159. Wu, Z. et al. METTL3 counteracts premature aging via m6A-dependent stabili- stellate cells and the progression of liver fibrosis. Biochem. Biophys. Res. Com-
zation of MIS12 mRNA. Nucleic Acids Res. 48, 11083–11096 (2020). mun. 463, 679–685 (2015).

Signal Transduction and Targeted Therapy (2022)7:374

 Epigenetic regulation of aging: implications for interventions of aging. . .
Wang et al.
19
192. Barutcu, A. R. et al. A TAD boundary is preserved upon deletion of the CTCF-rich 222. Fontan-Lozano, A. et al. Histone deacetylase inhibitors improve learning con-
Firre locus. Nat. Commun. 9, 1444 (2018). solidation in young and in KA-induced-neurodegeneration and SAMP-8-mutant
193. Konermann, S. et al. Genome-scale transcriptional activation by an engineered mice. Mol. Cell Neurosci. 39, 193–201 (2008).
CRISPR-Cas9 complex. Nature 517, 583–588 (2015). 223. Cabreiro, F. et al. Metformin retards aging in C. elegans by altering microbial
194. Yang, D., Yang, K. & Yang, M. in Aging and Aging-Related Diseases: Mechanisms folate and methionine metabolism. Cell 153, 228–239 (2013).
and Interventions (ed Wang, Z.) 17–35 (Springer Singapore, 2018). 224. Wang, Y. et al. Metformin exerts antidepressant effects by regulated DNA
195. Inzulza-Tapia, A. & Alarcon, M. Role of non-coding rna of human platelet in hydroxymethylation. Epigenomics 11, 655–667 (2019).
cardiovascular disease. Curr. Med. Chem. 29, 3420–3444 (2022). 225. Noren Hooten, N. et al. Metformin-mediated increase in DICER1 regulates
196. Wang, K. et al. Genomic profiling of native R loops with a DNA-RNA hybrid microRNA expression and cellular senescence. Aging Cell 15, 572–581 (2016).
recognition sensor. Sci. Adv. 7, eabe3516 (2021). 226. Zhang, S. et al. Metformin-induced microRNA-34a-3p downregulation alleviates
197. Yan, P. et al. Genome-wide R-loop landscapes during cell differentiation and senescence in human dental pulp stem cells by targeting CAB39 through the
reprogramming. Cell Rep. 32, 107870 (2020). AMPK/mTOR signaling pathway. Stem Cells Int. 2021, 6616240 (2021).
198. Nojima, T. et al. Deregulated expression of mammalian lncRNA through loss of 227. Martin-Montalvo, A. et al. Metformin improves healthspan and lifespan in mice.
SPT6 induces r-loop formation, replication stress, and cellular senescence. Mol Nat. Commun. 4, 2192 (2013).
Cell. 72, 970–984.e7 (2018). 228. Chen, M. et al. Metformin protects lens epithelial cells against senescence in a
199. Yaku, K., Okabe, K. & Nakagawa, T. NAD metabolism: implications in aging and naturally aged mouse model. Cell Death Discov. 8, 8 (2022).
longevity. Ageing Res. Rev. 47, 1–17 (2018). 229. Hong, S. et al. A high fat, sugar, and salt Western diet induces motor-muscular
200. Gu, X., Yao, H., Kwon, I. & Wang, G. Small-molecule activation of NAMPT as a and sensory dysfunctions and neurodegeneration in mice during aging: Ame-
potential neuroprotective strategy. Life Med. lnac012 https://doi.org/10.1093/ liorative action of metformin. CNS Neurosci. Ther. 27, 1458–1471 (2021).
lifemedi/lnac012 (2022). 230. Chen, Q., Thompson, J., Hu, Y. & Lesnefsky, E. J. Chronic metformin treatment
201. Shen, W. et al. Peroxisome proliferator-activated receptor γ coactivator 1α decreases cardiac injury during ischemia-reperfusion by attenuating endo-
maintains NAD+ bioavailability protecting against steatohepatitis. Life Med. plasmic reticulum stress with improved mitochondrial function. Aging (Albany
lnac031 https://doi.org/10.1093/lifemedi/lnac031 (2022). NY) 13, 7828–7845 (2021).
202. Fang, E. F. et al. NAD(+) augmentation restores mitophagy and limits acceler- 231. Anisimov, V. N. et al. If started early in life, metformin treatment increases life
ated aging in Werner syndrome. Nat. Commun. 10, 5284 (2019). span and postpones tumors in female SHR mice. Aging (Albany NY) 3, 148–157
203. Zhang, H. et al. NAD(+) repletion improves mitochondrial and stem cell function (2011).
and enhances life span in mice. Science 352, 1436–1443 (2016). 232. Smith, D. L. Jr. et al. Metformin supplementation and life span in Fischer-344
204. Romani, M. et al. NAD(+) boosting reduces age-associated amyloidosis and rats. J. Gerontol. A Biol. Sci. Med. Sci. 65, 468–474 (2010).
restores mitochondrial homeostasis in muscle. Cell Rep. 34, 108660 (2021). 233. Zhu, X. et al. Effect of metformin on cardiac metabolism and longevity in aged
205. Sasaki, L. et al. Intracrine activity involving NAD-dependent circadian steroido- female mice. Front. Cell Dev. Biol. 8, 626011 (2020).
genic activity governs age-associated meibomian gland dysfunction. Nat. Aging 234. Lu, M. et al. Metformin prevents dopaminergic neuron death in MPTP/P-induced
2, 105–114 (2022). mouse model of Parkinson’s disease via autophagy and mitochondrial ROS
206. Tarantini, S. et al. Nicotinamide mononucleotide (NMN) supplementation res- clearance. Int. J. Neuropsychopharmacol. 19, pyw047 (2016).
cues cerebromicrovascular endothelial function and neurovascular coupling 235. Yan, Q. et al. Activation of AMPK/mTORC1-mediated autophagy by metformin
responses and improves cognitive function in aged mice. Redox Biol. 24, 101192 reverses Clk1 deficiency-sensitized dopaminergic neuronal death. Mol. Pharm.
(2019). 92, 640–652 (2017).
207. Gong, B. et al. Nicotinamide riboside restores cognition through an upregulation 236. Wen, H. et al. Metformin and cyanidin 3-O-galactoside from Aronia melanocarpa
of proliferator-activated receptor-gamma coactivator 1alpha regulated beta- synergistically alleviate cognitive impairment in SAMP8 mice. Food Funct. 12,
secretase 1 degradation and mitochondrial gene expression in Alzheimer’s 10994–11008 (2021).
mouse models. Neurobiol. Aging 34, 1581–1588 (2013). 237. Kodali, M. et al. Metformin treatment in late middle age improves cognitive
208. Hou, Y. et al. NAD(+) supplementation normalizes key Alzheimer’s features and function with alleviation of microglial activation and enhancement of autop-
DNA damage responses in a new AD mouse model with introduced DNA repair hagy in the hippocampus. Aging Cell 20, e13277 (2021).
deficiency. Proc. Natl Acad. Sci. USA 115, E1876–E1885 (2018). 238. Lin, Y., Dai, X., Zhang, J. & Chen, X. Metformin alleviates the depression-like
209. Mitchell, S. J. et al. Nicotinamide improves aspects of healthspan, but not life- behaviors of elderly apoE4 mice via improving glucose metabolism and mito-
span, in mice. Cell Metab. 27, 667–676 e664 (2018). chondrial biogenesis. Behav. Brain Res. 423, 113772 (2022).
210. Bonkowski, M. S. & Sinclair, D. A. Slowing ageing by design: the rise of NAD(+) 239. Kim, H. et al. Metformin inhibits chronic kidney disease-induced DNA damage
and sirtuin-activating compounds. Nat. Rev. Mol. Cell Biol. 17, 679–690 (2016). and senescence of mesenchymal stem cells. Aging Cell 20, e13317 (2021).
211. Baur, J. A. et al. Resveratrol improves health and survival of mice on a high- 240. Yang, S. P. et al. Metformin ameliorates thymus degeneration of mice by reg-
calorie diet. Nature 444, 337–342 (2006). ulating mitochondrial function. Int. Immunopharmacol. 108, 108744 (2022).
212. Kornicka, K., Szlapka-Kosarzewska, J., Smieszek, A. & Marycz, K. 5-Azacytydine 241. Cai, H. et al. Metformin attenuates the D-galactose induced aging process via
and resveratrol reverse senescence and ageing of adipose stem cells via the UPR through the AMPK/ERK1/2 signaling pathways. Int. J. Mol. Med. 45,
modulation of mitochondrial dynamics and autophagy. J. Cell Mol. Med. 23, 715–730 (2020).
237–259 (2019). 242. Li, Q. et al. Obesity and hyperinsulinemia drive adipocytes to activate a cell cycle
213. Zhou, X. L. et al. SIRT1 activator (SRT1720) improves the follicle reserve and program and senesce. Nat. Med. 27, 1941–1953 (2021).
prolongs the ovarian lifespan of diet-induced obesity in female mice via acti- 243. Chen, Y. W., Harris, R. A., Hatahet, Z. & Chou, K. M. Ablation of XP-V gene causes
vating SIRT1 and suppressing mTOR signaling. J. Ovarian Res. 7, 97 (2014). adipose tissue senescence and metabolic abnormalities. Proc. Natl Acad. Sci. USA
214. Mitchell, S. J. et al. The SIRT1 activator SRT1720 extends lifespan and improves 112, E4556–E4564 (2015).
health of mice fed a standard diet. Cell Rep. 6, 836–843 (2014). 244. Zhu, Y. et al. Metformin treatment of juvenile mice alters aging-related devel-
215. Gano, L. B. et al. The SIRT1 activator SRT1720 reverses vascular endothelial opmental and metabolic phenotypes. Mech. Ageing Dev. 201, 111597 (2022).
dysfunction, excessive superoxide production, and inflammation with aging in 245. Strong, R. et al. Rapamycin-mediated mouse lifespan extension: Late-life dosage
mice. Am. J. Physiol. Heart Circ. Physiol. 307, H1754–H1763 (2014). regimes with sex-specific effects. Aging Cell 19, e13269 (2020).
216. Mercken, E. M. et al. SRT2104 extends survival of male mice on a standard diet 246. Siegmund, S. E. et al. Low-dose rapamycin extends lifespan in a mouse model of
and preserves bone and muscle mass. Aging Cell 13, 787–796 (2014). mtDNA depletion syndrome. Hum. Mol. Genet. 26, 4588–4605 (2017).
217. McIntyre, R. L. et al. From molecular promise to preclinical results: HDAC inhi- 247. Bitto, A. et al. Transient rapamycin treatment can increase lifespan and
bitors in the race for healthy aging drugs. EMBO Mol. Med. 11, e9854 (2019). healthspan in middle-aged mice. Elife. 5, e16351 (2016).
218. Majora, M. et al. HDAC inhibition improves autophagic and lysosomal function 248. Yin, Z. et al. Dietary restriction and rapamycin affect brain aging in mice by
to prevent loss of subcutaneous fat in a mouse model of Cockayne syndrome. attenuating age-related DNA methylation changes. Genes (Basel). 13, 699 (2022).
Sci. Transl. Med. 10, eaam7510 (2018). 249. Quarles, E. et al. Rapamycin persistently improves cardiac function in aged, male and
219. Jeong, M. Y. et al. Histone deacetylase activity governs diastolic dysfunction female mice, even following cessation of treatment. Aging Cell 19, e13086 (2020).
through a nongenomic mechanism. Sci. Transl. Med. 10, eaao0144 (2018). 250. Lesniewski, L. A. et al. Dietary rapamycin supplementation reverses age-related
220. Walsh, M. E. et al. The histone deacetylase inhibitor butyrate improves meta- vascular dysfunction and oxidative stress, while modulating nutrient-sensing,
bolism and reduces muscle atrophy during aging. Aging Cell 14, 957–970 (2015). cell cycle, and senescence pathways. Aging Cell 16, 17–26 (2017).
221. Fass, D. M. et al. Crebinostat: a novel cognitive enhancer that inhibits histone 251. Van Skike, C. E. et al. mTOR attenuation with rapamycin reverses neurovascular
deacetylase activity and modulates chromatin-mediated neuroplasticity. Neu- uncoupling and memory deficits in mice modeling Alzheimer’s disease. J.
ropharmacology 64, 81–96 (2013). Neurosci. 41, 4305–4320 (2021).

Signal Transduction and Targeted Therapy (2022)7:374

 Epigenetic regulation of aging: implications for interventions of aging. . .
Wang et al.
20
252. Towner, R. A. et al. Rapamycin restores brain vasculature, metabolism, and 283. Santos, E. L. et al. Long term treatment with ACE inhibitor enalapril decreases
blood-brain barrier in an inflammaging model. Geroscience 43, 563–578 body weight gain and increases life span in rats. Biochem. Pharm. 78, 951–958
(2021). (2009).
253. Lei, H., Wang, J., Ladiges, W. & Jiang, Z. Short-term oral rapamycin prevents age- 284. Graus-Nunes, F. et al. Differential effects of angiotensin receptor blockers on
related learning impairment in mice. Aging Pathobiol. Ther. 2, 166–167 (2020). pancreatic islet remodelling and glucose homeostasis in diet-induced obese
254. Singh, A. K. et al. Rapamycin confers neuroprotection against aging-induced mice. Mol. Cell Endocrinol. 439, 54–64 (2017).
oxidative stress, mitochondrial dysfunction, and neurodegeneration in old rats 285. Doeppner, T. R. et al. Long-term treatment with chloroquine increases life-
through activation of autophagy. Rejuvenation Res. 22, 60–70 (2019). span in middle-aged male mice possibly via autophagy modulation, protea-
255. Ham, D. J. et al. Distinct and additive effects of calorie restriction and rapamycin some inhibition and glycogen metabolism. Aging (Albany NY). 14, 4195–4210
in aging skeletal muscle. Nat. Commun. 13, 2025 (2022). (2022).
256. Kawakami, Y. et al. Rapamycin rescues age-related changes in muscle-derived 286. Strong, R. et al. Nordihydroguaiaretic acid and aspirin increase lifespan of
stem/progenitor cells from progeroid mice. Mol. Ther. Methods Clin. Dev. 14, genetically heterogeneous male mice. Aging Cell 7, 641–650 (2008).
64–76 (2019). 287. Liu, Z. et al. Cross-species metabolomic analysis identifies uridine as a potent
257. Garcia, D. N. et al. Effect of caloric restriction and rapamycin on ovarian aging in regeneration promoting factor. Cell Discov. 8, 6 (2022).
mice. Geroscience 41, 395–408 (2019). 288. Takahashi, K. & Yamanaka, S. Induction of pluripotent stem cells from mouse
258. Dou, X. et al. Short-term rapamycin treatment increases ovarian lifespan in embryonic and adult fibroblast cultures by defined factors. Cell 126, 663–676
young and middle-aged female mice. Aging Cell 16, 825–836 (2017). (2006).
259. Altschuler, R. A. et al. Rapamycin added to diet in late mid-life delays age-related 289. Takahashi, K. et al. Induction of pluripotent stem cells from adult human
hearing loss in UMHET4 mice. Front. Cell Neurosci. 15, 658972 (2021). fibroblasts by defined factors. Cell 131, 861–872 (2007).
260. Altschuler, R. A. et al. Rapamycin but not acarbose decreases age-related loss of 290. Koche, R. P. et al. Reprogramming factor expression initiates widespread tar-
outer hair cells in the mouse Cochlea. Hear Res. 370, 11–15 (2018). geted chromatin remodeling. Cell Stem Cell 8, 96–105 (2011).
261. An, J. Y. et al. Rapamycin rejuvenates oral health in aging mice. Elife. 9, e54318 291. Lu, Y. et al. Reprogramming to recover youthful epigenetic information and
(2020). restore vision. Nature 588, 124–129 (2020).
262. An, J. Y. et al. Rapamycin treatment attenuates age-associated periodontitis in 292. Abad, M. et al. Reprogramming in vivo produces teratomas and iPS cells with
mice. Geroscience 39, 457–463 (2017). totipotency features. Nature 502, 340–345 (2013).
263. Deblon, N. et al. Chronic mTOR inhibition by rapamycin induces muscle insulin 293. Manukyan, M. & Singh, P. B. Epigenome rejuvenation: HP1β mobility as a
resistance despite weight loss in rats. Br. J. Pharm. 165, 2325–2340 (2012). measure of pluripotent and senescent chromatin ground states. Sci. Rep. 4, 4789
264. Kim, Y., Lee, J. S. & Joo, Y. H. Rapamycin increases the incidence of neu- (2014).
ropsychiatric illness in kidney transplant patients through the suppression of 294. Olova, N., Simpson, D. J., Marioni, R. E. & Chandra, T. Partial reprogramming
neural stem cells. Transl. Psychiatry 10, 156 (2020). induces a steady decline in epigenetic age before loss of somatic identity. Aging
265. Liu, J. et al. Delay in oocyte aging in mice by the antioxidant N-acetyl-L-cysteine Cell 18, e12877 (2019).
(NAC). Hum. Reprod. 27, 1411–1420 (2012). 295. Sarkar, T. J. et al. Transient non-integrative expression of nuclear reprogram-
266. Garg, G., Singh, S., Singh, A. K. & Rizvi, S. I. N-acetyl-l-cysteine attenuates oxi- ming factors promotes multifaceted amelioration of aging in human cells. Nat.
dative damage and neurodegeneration in rat brain during aging. Can. J. Physiol. Commun. 11, 1545 (2020).
Pharm. 96, 1189–1196 (2018). 296. Gill, D. et al. Multi-omic rejuvenation of human cells by maturation phase
267. Flurkey, K., Astle, C. M. & Harrison, D. E. Life extension by diet restriction and N- transient reprogramming. Elife. 11, e71624 (2022).
acetyl-L-cysteine in genetically heterogeneous mice. J. Gerontol. A Biol. Sci. Med. 297. Ocampo, A. et al. In vivo amelioration of age-associated hallmarks by partial
Sci. 65, 1275–1284 (2010). reprogramming. Cell 167, 1719–1733.e1712 (2016).
268. Costa, M. et al. N-acetylcysteine protects memory decline induced by strepto- 298. Alle, Q. et al. A single short reprogramming early in life improves fitness and
zotocin in mice. Chem. Biol. Interact. 253, 10–17 (2016). increases lifespan in old age. Preprint at bioRxiv https://doi.org/10.1111/
269. More, J. et al. N-acetylcysteine prevents the spatial memory deficits and the acel.13714 (2021).
redox-dependent RyR2 decrease displayed by an Alzheimer’s disease rat model. 299. Chondronasiou, D. et al. Multi-omic rejuvenation of naturally aged tissues by a
Front. Aging Neurosci. 10, 399 (2018). single cycle of transient reprogramming. Aging Cell 21, e13578 (2022).
270. Zhu, Q. Y. et al. N-acetyl cysteine ameliorates aortic fibrosis by promoting M2 300. Roos, C. M. et al. Chronic senolytic treatment alleviates established vasomotor
macrophage polarization in aging mice. Redox Rep. 26, 170–175 (2021). dysfunction in aged or atherosclerotic mice. Aging Cell 15, 973–977 (2016).
271. Meryk, A. et al. Antioxidants N-acetylcysteine and vitamin C improve T cell 301. Lewis-McDougall, F. C. et al. Aged-senescent cells contribute to impaired heart
commitment to memory and long-term maintenance of immunological mem- regeneration. Aging Cell 18, e12931 (2019).
ory in old mice. Antioxidants (Basel). 9, 1152 (2020). 302. Zhou, Y. et al. Senolytics improve bone forming potential of bone marrow
272. Kawaguchi, K., Hashimoto, M. & Sugimoto, M. An antioxidant suppressed lung mesenchymal stem cells from aged mice. NPJ Regen. Med. 6, 34 (2021).
cellular senescence and enhanced pulmonary function in aged mice. Biochem. 303. Farr, J. N. et al. Targeting cellular senescence prevents age-related bone loss in
Biophys. Res. Commun. 541, 43–49 (2021). mice. Nat. Med. 23, 1072–1079 (2017).
273. Zhou, X. et al. Suppression effect of N-acetylcysteine on bone loss in ovar- 304. Novais, E. J. et al. Long-term treatment with senolytic drugs Dasatinib and
iectomized mice. Am. J. Transl. Res. 12, 731–742 (2020). Quercetin ameliorates age-dependent intervertebral disc degeneration in mice.
274. Marie, A. et al. N-acetylcysteine treatment reduces age-related hearing loss and Nat. Commun. 12, 5213 (2021).
memory impairment in the senescence-accelerated prone 8 (SAMP8) mouse 305. Zhang, P. et al. Senolytic therapy alleviates Abeta-associated oligodendrocyte
model. Aging Dis. 9, 664–673 (2018). progenitor cell senescence and cognitive deficits in an Alzheimer’s disease
275. Miller, R. A. et al. Canagliflozin extends life span in genetically heterogeneous model. Nat. Neurosci. 22, 719–728 (2019).
male but not female mice. JCI Insight. 5, e140019 (2020). 306. Ogrodnik, M. et al. Obesity-induced cellular senescence drives anxiety and
276. Harrison, D. E. et al. Acarbose improves health and lifespan in aging HET3 mice. impairs neurogenesis. Cell Metab. 29, 1061–1077 e1068 (2019).
Aging Cell 18, e12898 (2019). 307. Lee, G. et al. SREBP1c-PARP1 axis tunes anti-senescence activity of adipocytes
277. Strong, R. et al. Longer lifespan in male mice treated with a weakly estrogenic and ameliorates metabolic imbalance in obesity. Cell Metab. 34, 702–718 e705
agonist, an antioxidant, an α‐glucosidase inhibitor or a Nrf2‐inducer. Aging Cell (2022).
15, 872–884 (2016). 308. Wang, L. et al. Targeting p21(Cip1) highly expressing cells in adipose tissue
278. Harrison, D. E. et al. Acarbose, 17-alpha-estradiol, and nordihydroguaiaretic acid alleviates insulin resistance in obesity. Cell Metab. 34, 75–89 e78 (2022).
extend mouse lifespan preferentially in males. Aging Cell 13, 273–282 (2014). 309. Justice, J. N. et al. Senolytics in idiopathic pulmonary fibrosis: Results from a first-
279. Geng, L. et al. Chemical screen identifies a geroprotective role of quercetin in in-human, open-label, pilot study. EBioMedicine 40, 554–563 (2019).
premature aging. Protein Cell 10, 417–435 (2019). 310. Schafer, M. J. et al. Cellular senescence mediates fibrotic pulmonary disease. Nat.
280. Geng, L. et al. Low-dose quercetin positively regulates mouse healthspan. Pro- Commun. 8, 14532 (2017).
tein Cell 10, 770–775 (2019). 311. Dungan, C. M. et al. Deletion of SA beta-Gal+ cells using senolytics improves
281. Ma, S. et al. Single-cell transcriptomic atlas of primate cardiopulmonary aging. muscle regeneration in old mice. Aging Cell 21, e13528 (2022).
Cell Res. 31, 415–432 (2021). 312. Krzystyniak, A. et al. Combination of dasatinib and quercetin improves cognitive
282. Zakaria, A., Hamdi, N. & Abdel-Kader, R. M. Methylene blue improves brain abilities in aged male Wistar rats, alleviates inflammation and changes hippo-
mitochondrial ABAD functions and decreases abeta in a neuroinflammatory campal synaptic plasticity and histone H3 methylation profile. Aging (Albany NY)
Alzheimer’s disease mouse model. Mol. Neurobiol. 53, 1220–1228 (2016). 14, 572–595 (2022).

Signal Transduction and Targeted Therapy (2022)7:374

 Epigenetic regulation of aging: implications for interventions of aging. . .
Wang et al.
21
313. Spinelli, R. et al. ZMAT3 hypomethylation contributes to early senescence of 342. Barrès, R. et al. Acute exercise remodels promoter methylation in human skeletal
preadipocytes from healthy first-degree relatives of type 2 diabetics. Aging Cell muscle. Cell Metab. 15, 405–411 (2012).
21, e13557 (2022). 343. Nitert, M. D. et al. Impact of an exercise intervention on DNA methylation in
314. Kim, H. et al. Inhibition of matrix metalloproteinase expression by selective skeletal muscle from first-degree relatives of patients with type 2 diabetes.
clearing of senescent dermal fibroblasts attenuates ultraviolet-induced pho- Diabetes 61, 3322–3332 (2012).
toaging. Biomed. Pharmacother. 150, 113034 (2022). 344. Gomez-Pinilla, F. et al. Exercise impacts brain-derived neurotrophic factor
315. Chang, J. et al. Clearance of senescent cells by ABT263 rejuvenates aged plasticity by engaging mechanisms of epigenetic regulation. Eur. J. Neurosci. 33,
hematopoietic stem cells in mice. Nat. Med. 22, 78–83 (2016). 383–390 (2011).
316. Baar, M. P. et al. Targeted apoptosis of senescent cells restores tissue home- 345. Egan, B. et al. Exercise intensity-dependent regulation of peroxisome
ostasis in response to chemotoxicity and aging. Cell 169, 132–147 e116 (2017). proliferator-activated receptor coactivator-1 mRNA abundance is associated
317. Fuhrmann-Stroissnigg, H. et al. Identification of HSP90 inhibitors as a novel class with differential activation of upstream signalling kinases in human skeletal
of senolytics. Nat. Commun. 8, 422 (2017). muscle. J. Physiol. 588, 1779–1790 (2010).
318. Triana-Martinez, F. et al. Identification and characterization of cardiac glycosides 346. Ogasawara, R. et al. MicroRNA expression profiling in skeletal muscle reveals
as senolytic compounds. Nat. Commun. 10, 4731 (2019). different regulatory patterns in high and low responders to resistance training.
319. Weindruch, R., Walford, R. L., Fligiel, S. & Guthrie, D. The retardation of aging in Physiol. Genomics 48, 320–324 (2016).
mice by dietary restriction: longevity, cancer, immunity and lifetime energy 347. Fyfe, J. J. et al. Concurrent exercise incorporating high-intensity interval or
intake. J. Nutr. 116, 641–654 (1986). continuous training modulates mTORC1 signaling and microRNA expression in
320. Hahn, O. et al. Dietary restriction protects from age-associated DNA methylation human skeletal muscle. Am. J. Physiol. Regul. Integr. Comp. Physiol. 310,
and induces epigenetic reprogramming of lipid metabolism. Genome Biol. 18, R1297–R1311 (2016).
56 (2017). 348. van Praag, H., Shubert, T., Zhao, C. & Gage, F. H. Exercise enhances learning and
321. Mitchell, S. J. et al. Effects of sex, strain, and energy intake on hallmarks of aging hippocampal neurogenesis in aged mice. J. Neurosci. 25, 8680–8685 (2005).
in mice. Cell Metab. 23, 1093–1112 (2016). 349. Rubenstein, A. B. et al. Skeletal muscle transcriptome response to a bout of
322. Hepple, R. T., Baker, D. J., Kaczor, J. J. & Krause, D. J. Long-term caloric restriction endurance exercise in physically active and sedentary older adults. Am. J. Phy-
abrogates the age-related decline in skeletal muscle aerobic function. FASEB J. siol. Endocrinol. Metab. 322, E260–e277 (2022).
19, 1320–1322 (2005). 350. Ruple, B. A. et al. Resistance training rejuvenates the mitochondrial methylome
323. Rippe, C. et al. Short-term calorie restriction reverses vascular endothelial dys- in aged human skeletal muscle. FASEB J. 35, e21864 (2021).
function in old mice by increasing nitric oxide and reducing oxidative stress. 351. Blocquiaux, S. et al. Recurrent training rejuvenates and enhances transcriptome
Aging Cell 9, 304–312 (2010). and methylome responses in young and older human muscle. JCSM Rapid
324. Valdez, G. et al. Attenuation of age-related changes in mouse neuromuscular Commun. 5, 10–32 (2022).
synapses by caloric restriction and exercise. Proc. Natl Acad. Sci. USA 107, 352. Dorling, J. L. et al. Effects of caloric restriction on human physiological, psy-
14863–14868 (2010). chological, and behavioral outcomes: highlights from CALERIE phase 2. Nutr.
325. Cerletti, M. et al. Short-term calorie restriction enhances skeletal muscle stem Rev. 79, 98–113 (2021).
cell function. Cell Stem Cell 10, 515–519 (2012). 353. Ravussin, E. et al. A 2-year randomized controlled trial of human caloric
326. Cohen, H. Y. et al. Calorie restriction promotes mammalian cell survival by restriction: feasibility and effects on predictors of health span and longevity. J.
inducing the SIRT1 deacetylase. Science 305, 390–392 (2004). Gerontol. A Biol. Sci. Med. Sci. 70, 1097–1104 (2015).
327. Ma, S. et al. Caloric restriction reprograms the single-cell transcriptional land- 354. Schroder, J. D. et al. Effects of time-restricted feeding in weight loss, meta-
scape of Rattus Norvegicus aging. Cell 180, 984–1001 e1022 (2020). bolic syndrome and cardiovascular risk in obese women. J. Transl. Med. 19, 3
328. Colman, R. J. et al. Caloric restriction delays disease onset and mortality in (2021).
rhesus monkeys. Science 325, 201–204 (2009). 355. Wilkinson, M. J. et al. Ten-hour time-restricted eating reduces weight, blood
329. Maegawa, S. et al. Caloric restriction delays age-related methylation drift. Nat. pressure, and atherogenic lipids in patients with metabolic syndrome. Cell
Commun. 8, 539 (2017). Metab. 31, 92–104 e105 (2020).
330. Rochon, J. et al. Design and conduct of the CALERIE study: comprehensive 356. Sutton, E. F. et al. Early time-restricted feeding improves insulin sensitivity, blood
assessment of the long-term effects of reducing intake of energy. J. Gerontol. A pressure, and oxidative stress even without weight loss in men with pre-
Biol. Sci. Med. Sci. 66, 97–108 (2011). diabetes. Cell Metab. 27, 1212–1221 e1213 (2018).
331. Rickman, A. D. et al. The CALERIE Study: design and methods of an innovative 357. Rong, S. et al. Association of skipping breakfast with cardiovascular and all-
25% caloric restriction intervention. Contemp. Clin. Trials 32, 874–881 (2011). cause mortality. J. Am. Coll. Cardiol. 73, 2025–2032 (2019).
332. Belsky, D. W. et al. Change in the rate of biological aging in response to caloric 358. Longo, V. D. & Anderson, R. M. Nutrition, longevity and disease: from molecular
restriction: CALERIE biobank analysis. J. Gerontol. A Biol. Sci. Med. Sci. 73, 4–10 mechanisms to interventions. Cell 185, 1455–1470 (2022).
(2017). 359. Wei, M. et al. Fasting-mimicking diet and markers/risk factors for aging, dia-
333. Dorling, J. L. et al. Effect of 2 years of calorie restriction on liver biomarkers: betes, cancer, and cardiovascular disease. Sci. Transl. Med. 9, eaai8700 (2017).
results from the CALERIE phase 2 randomized controlled trial. Eur. J. Nutr. 60, 360. Mansur, A. P. et al. Serum concentrations and gene expression of sirtuin 1 in
1633–1643 (2021). healthy and slightly overweight subjects after caloric restriction or resveratrol
334. Kraus, W. E. et al. 2 years of calorie restriction and cardiometabolic risk (CAL- supplementation: a randomized trial. Int. J. Cardiol. 227, 788–794 (2017).
ERIE): exploratory outcomes of a multicentre, phase 2, randomised controlled 361. Lilja, S. et al. Five days periodic fasting elevates levels of longevity related
trial. Lancet Diabetes Endocrinol. 7, 673–683 (2019). christensenella and sirtuin expression in humans. Int. J. Mol. Sci. 22, 2331 (2021).
335. Martin, C. K. et al. Effect of calorie restriction on mood, quality of life, sleep, and 362. Fitzgerald, K. N. et al. Potential reversal of epigenetic age using a diet and
sexual function in healthy nonobese adults: The CALERIE 2 randomized clinical lifestyle intervention: a pilot randomized clinical trial. Aging (Albany NY) 13,
trial. JAMA Intern. Med. 176, 743–752 (2016). 9419–9432 (2021).
336. Redman, L. M. et al. Metabolic slowing and reduced oxidative damage with 363. Fiorito, G. et al. DNA methylation-based biomarkers of aging were slowed down
sustained caloric restriction support the rate of living and oxidative damage in a two-year diet and physical activity intervention trial: the DAMA study. Aging
theories of aging. Cell Metab. 27, 805–815.e804 (2018). Cell 20, e13439 (2021).
337. Acosta-Rodríguez, V. A., Rijo-Ferreira, F., Green, C. B. & Takahashi, J. S. Impor- 364. Lee, D. C. et al. Running as a key lifestyle medicine for longevity. Prog. Cardio-
tance of circadian timing for aging and longevity. Nat. Commun. 12, 2862 vasc. Dis. 60, 45–55 (2017).
(2021). 365. Chakravarty, E. F., Hubert, H. B., Lingala, V. B. & Fries, J. F. Reduced disability and
338. Solanas, G. et al. Aged stem cells reprogram their daily rhythmic functions to mortality among aging runners: a 21-year longitudinal study. Arch. Intern. Med.
adapt to stress. Cell 170, 678–692.e620 (2017). 168, 1638–1646 (2008).
339. Hor, C. N. et al. Sleep-wake-driven and circadian contributions to daily rhythms 366. Moore, S. C. et al. Leisure time physical activity of moderate to vigorous intensity
in gene expression and chromatin accessibility in the murine cortex. Proc. Natl and mortality: a large pooled cohort analysis. PLoS Med. 9, e1001335 (2012).
Acad. Sci. USA 116, 25773–25783 (2019). 367. Hieda, M. et al. One-year committed exercise training reverses abnormal left
340. Libert, S. et al. Deviation of innate circadian period from 24 h reduces longevity ventricular myocardial stiffness in patients with stage b heart failure with pre-
in mice. Aging Cell 11, 794–800 (2012). served ejection fraction. Circulation 144, 934–946 (2021).
341. Morris, C. J., Purvis, T. E., Hu, K. & Scheer, F. A. Circadian misalignment increases 368. Kirk, B. et al. Leucine-enriched whey protein supplementation, resistance-based
cardiovascular disease risk factors in humans. Proc. Natl Acad. Sci. USA 113, exercise, and cardiometabolic health in older adults: a randomized controlled
E1402–E1411 (2016). trial. J. Cachexia Sarcopenia Muscle 12, 2022–2033 (2021).

Signal Transduction and Targeted Therapy (2022)7:374

 Epigenetic regulation of aging: implications for interventions of aging. . .
Wang et al.
22
369. Howden, E. J. et al. Reversing the cardiac effects of sedentary aging in middle 390. Dollerup, O. L. et al. Effects of nicotinamide riboside on endocrine pancreatic
age-a Randomized controlled trial: implications for heart failure prevention. function and incretin hormones in nondiabetic men with obesity. J. Clin.
Circulation 137, 1549–1560 (2018). Endocrinol. Metab. 104, 5703–5714 (2019).
370. Sandri, M. et al. Age-related effects of exercise training on diastolic function in 391. Dollerup, O. L. et al. A randomized placebo-controlled clinical trial of nicotina-
heart failure with reduced ejection fraction: the Leipzig Exercise Intervention in mide riboside in obese men: safety, insulin-sensitivity, and lipid-mobilizing
Chronic Heart Failure and Aging (LEICA) Diastolic Dysfunction Study. Eur. Heart J. effects. Am. J. Clin. Nutr. 108, 343–353 (2018).
33, 1758–1768 (2012). 392. Dai, H., Sinclair, D. A., Ellis, J. L. & Steegborn, C. Sirtuin activators and inhibitors:
371. Kitzman, D. W. et al. Effect of endurance exercise training on endothelial promises, achievements, and challenges. Pharm. Ther. 188, 140–154 (2018).
function and arterial stiffness in older patients with heart failure and preserved 393. Venkatasubramanian, S. et al. Cardiovascular effects of a novel SIRT1 activator,
ejection fraction: a randomized, controlled, single-blind trial. J. Am. Coll. Cardiol. SRT2104, in otherwise healthy cigarette smokers. J. Am. Heart Assoc. 2, e000042
62, 584–592 (2013). (2013).
372. Ngandu, T. et al. A 2 year multidomain intervention of diet, exercise, cognitive 394. Libri, V. et al. A pilot randomized, placebo controlled, double blind phase I trial
training, and vascular risk monitoring versus control to prevent cognitive of the novel SIRT1 activator SRT2104 in elderly volunteers. PLoS ONE 7, e51395
decline in at-risk elderly people (FINGER): a randomised controlled trial. Lancet (2012).
385, 2255–2263 (2015). 395. Baksi, A. et al. A phase II, randomized, placebo-controlled, double-blind, multi-
373. ten Brinke, L. F. et al. Aerobic exercise increases hippocampal volume in older dose study of SRT2104, a SIRT1 activator, in subjects with type 2 diabetes. Br. J.
women with probable mild cognitive impairment: a 6-month randomised Clin. Pharm. 78, 69–77 (2014).
controlled trial. Br. J. Sports Med. 49, 248–254 (2015). 396. Sands, B. E. et al. Assessing colonic exposure, safety, and clinical activity of
374. Madden, K. M. et al. Short-term aerobic exercise reduces arterial stiffness in SRT2104, a novel oral SIRT1 activator, in patients with mild to moderate
older adults with type 2 diabetes, hypertension, and hypercholesterolemia. ulcerative colitis. Inflamm. Bowel Dis. 22, 607–614 (2016).
Diabetes Care 32, 1531–1535 (2009). 397. Knowler, W. C. et al. Reduction in the incidence of type 2 diabetes with lifestyle
375. Lanza, I. R. et al. Endurance exercise as a countermeasure for aging. Diabetes 57, intervention or metformin. N. Engl. J. Med. 346, 393–403 (2002).
2933–2942 (2008). 398. Hong, J. et al. Effects of metformin versus glipizide on cardiovascular outcomes
376. Short, K. R. et al. Impact of aerobic exercise training on age-related changes in in patients with type 2 diabetes and coronary artery disease. Diabetes Care 36,
insulin sensitivity and muscle oxidative capacity. Diabetes 52, 1888–1896 (2003). 1304–1311 (2013).
377. Johnson, M. L. et al. Differential effect of endurance training on mitochondrial 399. Wang, C. P., Lorenzo, C. & Espinoza, S. E. Frailty attenuates the impact of met-
protein damage, degradation, and acetylation in the context of aging. J. Ger- formin on reducing mortality in older adults with type 2 diabetes. J. Endocrinol.
ontol. A Biol. Sci. Med. Sci. 70, 1386–1393 (2015). Diabetes Obes. 2, 1031 (2014).
378. Hooshmand-Moghadam, B. et al. The effect of 12-week resistance exercise 400. Luchsinger, J. A. et al. Metformin in amnestic mild cognitive impairment: results
training on serum levels of cellular aging process parameters in elderly men. of a pilot randomized placebo controlled clinical trial. J. Alzheimers Dis. 51,
Exp. Gerontol. 141, 111090 (2020). 501–514 (2016).
379. Olioso, D. et al. Effects of aerobic and resistance training on circulating micro- 401. de Kreutzenberg, S. V. et al. Metformin improves putative longevity effectors in
rna expression profile in subjects with type 2 diabetes. J. Clin. Endocrinol. Metab. peripheral mononuclear cells from subjects with prediabetes. A randomized
104, 1119–1130 (2019). controlled trial. Nutr. Metab. Cardiovasc. Dis. 25, 686–693 (2015).
380. Akbarinia, A., Kargarfard, M. & Naderi, M. Aerobic training improves platelet 402. Mannick, J. B. et al. mTOR inhibition improves immune function in the elderly.
function in type 2 diabetic patients: role of microRNA-130a and GPIIb. Acta Sci. Transl. Med. 6, 268ra179 (2014).
Diabetol. 55, 893–899 (2018). 403. Mannick, J. B. et al. TORC1 inhibition enhances immune function and reduces
381. Taghizadeh, M. et al. Long-term aerobic exercise training in type two diabetic infections in the elderly. Sci. Transl. Med. 10, eaaq1564 (2018).
patients alters the expression of miRNA-223 and its corresponding target, the 404. Kraig, E. et al. A randomized control trial to establish the feasibility and safety of
P2RY12 receptor, attenuating platelet function. Clin. Hemorheol. Microcirc. 80, rapamycin treatment in an older human cohort: immunological, physical per-
107–116 (2022). formance, and cognitive effects. Exp. Gerontol. 105, 53–69 (2018).
382. Yoshino, M. et al. Nicotinamide mononucleotide increases muscle insulin sen- 405. Singh, M. et al. Effect of low-dose rapamycin on senescence markers and
sitivity in prediabetic women. Science 372, 1224–1229 (2021). physical functioning in older adults with coronary artery disease: results of a
383. Igarashi, M. et al. Chronic nicotinamide mononucleotide supplementation ele- pilot study. J. Frailty Aging 5, 204–207 (2016).
vates blood nicotinamide adenine dinucleotide levels and alters muscle func- 406. Hickson, L. J. et al. Senolytics decrease senescent cells in humans: Preliminary
tion in healthy older men. NPJ Aging 8, 5 (2022). report from a clinical trial of Dasatinib plus Quercetin in individuals with diabetic
384. Kim, M. et al. Effect of 12-week intake of nicotinamide mononucleotide on sleep kidney disease. EBioMedicine 47, 446–456 (2019).
quality, fatigue, and physical performance in older Japanese adults: a rando-
mized, double-blind placebo-controlled study. Nutrients 14, 755 (2022).
385. Zhou, B. et al. Boosting NAD level suppresses inflammatory activation of PBMCs Open Access This article is licensed under a Creative Commons
in heart failure. J. Clin. Invest. 130, 6054–6063 (2020). Attribution 4.0 International License, which permits use, sharing,
386. Brakedal, B. et al. The NADPARK study: a randomized phase I trial of nicotina- adaptation, distribution and reproduction in any medium or format, as long as you give
mide riboside supplementation in Parkinson’s disease. Cell Metab. 34, 396–407 appropriate credit to the original author(s) and the source, provide a link to the Creative
e396 (2022). Commons license, and indicate if changes were made. The images or other third party
387. Martens, C. R. et al. Chronic nicotinamide riboside supplementation is well- material in this article are included in the article’s Creative Commons license, unless
tolerated and elevates NAD(+) in healthy middle-aged and older adults. Nat. indicated otherwise in a credit line to the material. If material is not included in the
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388. Remie, C. M. E. et al. Nicotinamide riboside supplementation alters body com- regulation or exceeds the permitted use, you will need to obtain permission directly
position and skeletal muscle acetylcarnitine concentrations in healthy obese from the copyright holder. To view a copy of this license, visit http://
humans. Am. J. Clin. Nutr. 112, 413–426 (2020). creativecommons.org/licenses/by/4.0/.
389. Dollerup, O. L. et al. Nicotinamide riboside does not alter mitochondrial
respiration, content or morphology in skeletal muscle from obese and insulin-
resistant men. J. Physiol. 598, 731–754 (2020). © The Author(s) 2022

Signal Transduction and Targeted Therapy (2022)7:374