Biomedicine & Pharmacotherapy 156 (2022) 113984

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Biomedicine & Pharmacotherapy

journal homepage: www.elsevier.com/locate/biopha

Review

Novel histone post-translational modifications in diabetes and

complications of diabetes: The underlying mechanisms and implications
Dongze Li a, b, c, 1, Li Zhang d, 1, Yanqiu He a, b, c, 1, Tingting Zhou a, b, c, 1, Xi Cheng a, b, c, 1,
Wei Huang 1, a, b, c, *, Yong Xu 1, a, b, c, *
a
Department of Endocrinology and Metabolism, Metabolic Vascular Disease Key Laboratory of Sichuan Province, The Affiliated Hospital of Southwest Medical
University, Luzhou 646000, Sichuan, China
b
Sichuan Clinical Research Center for Nephropathy, Luzhou 646000, Sichuan, China
c
Department of Endocrinology and Metabolism, The Affiliated Hospital of Southwest Medical University, Luzhou 646000, Sichuan, China
d
Department of Vascular Surgery, The Affiliated Hospital of Southwest Medical University, Luzhou 646000, Sichuan, China

A R T I C L E I N F O A B S T R A C T

Keywords: Diabetes is a group of global epidemic diseases with high prevalence and morbidity. The pathophysiological
Diabetes changes of diabetes can be affected by genes, environmental factors, and several socioeconomic factors. Histone
Epigenetics post-translational modification is the central epigenetic mechanism regulated by genetic and environmental
Acylation
factors. Histone methylation and acetylation have been thoroughly studied in various pathological processes of
Histone modification
Metabolic memory
diabetes, such as the metabolic memory phenomenon, inflammation, and endothelial dysfunction. In recent
years, an increasing number of novel histone acylation modifications have been identified by mass spectrometry.
The level of any novel histone modification depends on the relative concentration of its respective acyl-CoA,
which can be generated through various intermediate metabolic pathways. Although studies have shown that
these novel modifications and metabolic regulation of histone acylation have a close connection with diabetes,
investigations on this connection have lagged. Moreover, the locations of acylation modification sites and how
enzymatic reactions regulate diabetes and its complications remain largely unknown. This review summarizes
the relationship between novel histone modifications and diabetes, mainly focusing on β-hydroxybutyrylation,
propionylation, butyrylation, malonylation, and succinylation. We aim to highlight the typical characteristics of
novel acylation modifications and provide a new perspective for further research on the pathogenesis and
treatment of diabetes and its complications.

1. Introduction
It is estimated that 537 million adults worldwide are living with
diabetes according to the latest International Diabetes Federation (IDF)
report. Chronic complications of diabetes caused by long-term chronic
hyperglycemia, often affecting the cardiovascular system, nervous sys­
tem, and urinary system, are the main source of the health and economic
burdens of patients. The pathogenesis of diabetes is complex, involving
genetic susceptibility and environmental factors. Emerging evidence
derived from Diabetes Control and Complications Trial (DCCT) and
Epidemiology of Diabetes Interventions and Complications Study (EDIC)
[1] demonstrates a “metabolic memory” phenomenon: early exposure to
hyperglycemia results in persistently induced damage even after the
hyperglycemic state is under control, which indicates that epigenetics is
a critical factor as a bridge between genes and the environment.
The basic goal of epigenetics is to identify heritable changes in gene
expression or cellular phenotype with no accompanying alterations in
the DNA sequence. Epigenetic mechanisms principally include histone

Abbreviations: HPTMs, Histones Post-translational Modifications; CoA, coenzyme A; Kac, lysine acetylation; Kme, lysine methylation; Kpr, lysine propionylation;
Kbu, lysine butyrylation; Ksucc, lysine succinylation; Kma, lysine malonylation; Kbhb, lysine β-hydroxybutyrylation; Kbz, lysine benzoylation; HDAC, histone
deacetylase; KD, ketogenic diet; BHB, β-hydroxybutyrate; DKD, diabetic kidney disease; SCFAs, short chain fatty acids; TNF-α, tumor necrosis factor alpha; MCP-1,
macrophage chemoattractant protein-1; IL, interleukin; VEGF, vascular endothelial growth factor; MMP-2, matrix metalloproteinase-2.
* Corresponding authors at: Department of Endocrinology and Metabolism, Metabolic Vascular Disease Key Laboratory of Sichuan Province, The Affiliated Hospital
of Southwest Medical University, Luzhou 646000, Sichuan, China.
E-mail addresses: huangwei1212520@163.com (W. Huang), xywyll@swmu.edu.cn (Y. Xu).
1
These authors have contributed equally to this work and share first authorship.

https://doi.org/10.1016/j.biopha.2022.113984

Available online 7 November 2022

0753-3322/© 2022 The Authors. Published by Elsevier Masson SAS. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
D. Li et al.
post-transcriptional modifications (HPTMs), DNA methylation, and
small non-coding regulatory RNAs [2]. Histones can bind tightly to DNA
to form nucleosome structures, including core histones H2A, H2B, H3,
H4, and a linker histone H1 [3]. HPTM refers to a covalent process in
which different acylation modifications are added to one or several
amino acid residues on the tails of histones, which alters the loose or
tight binding state between histones and DNA and effectively regulates
the transcriptional activity of the target gene. The most prominent of
HPTMs are acetylation (Kac) and methylation (Kme). In recent years,
benefiting from the development of anti-acylated antibodies and the use
of ultrasensitive mass spectrometry (MS), types of novel histone lysine
acylations have been identified and characterized, such as lysine pro­
pionylation (Kpr), butyrylation (Kbu) [4], succinylation (Ksucc) [5],
crotonylation (Kcr) [6], malonylation (Kma) [7], 2-hydroxyisobutyla­
tion (Khib) [8], glutarylation (Kglu) [9], β-hydroxybutyrylation
(Kbhb) [10], benzoylation (Kbz) [11], and, most recently, lactylation
(Kla) [12]. These external modifications constitute the basic elements of
the histone code. Acyl-CoA is not only a specific donor of these histone
modifications but also a metabolic intermediate in many metabolic
pathways. In addition to some endogenous acyl-CoA (such as acetyl-CoA
and succinyl-CoA) produced by glucose and lipid metabolism in healthy
or diabetic states, acyl-CoA synthetase (ACS), also known as acyl-CoA
synthetase short chain member 2 (ACSS2), can promote the produc­
tion of acyl-CoA (such as acetyl-CoA, propionyl-CoA, butyryl-CoA, and
β-hydroxybutyryl-CoA).
Many studies have shown that the development of diabetes and the
occurrence of the “metabolic memory” phenomenon is closely related to
classic histone modifications [13], and recent studies also have hinted at
a relationship between diabetes and some novel HPTMs, including
β-hydroxybutyrylation, propionylation, butyrylation, malonylation, and
succinylation.
In this review, we briefly outline the relationship between diabetes
and classic HPTMs and summarize the latest advances of the above-
mentioned novel HPTMs in diabetes and complications of diabetes.
Moreover, we detail the traits of these novel HPTMs, including their
acylation donors, sites, and corresponding enzymes, and we briefly
analyze the less common HPTMs. We also provide a fresh perspective on
the pathogenesis of diabetes and its complications and suggest possible
strategies for diabetes prevention and anti-diabetic therapies based on
novel HPTMs pathways.
2. General overview of histone modifications in diabetes and
complications of diabetes
Due to the inherent complexity of the pathogenesis of diabetes and
its complications, much effort has been put towards determining the
underlying etiologies. In recent decades, accumulating evidence has
shown that HPTMs play a significant role in modulating gene expression
under the diabetic state and causing the phenomenon of “metabolic
memory.”
Acetylation and methylation are the two classic and most thoroughly
studied histone modifications related to diabetes. A large volume of data
has confirmed that these histone modifications are intimately related
with the “metabolic memory” phenomenon [14], inflammatory
response [15], endothelial dysfunction [16], and other pathological
processes of diabetes. For example, compared with non-diabetic pa­
tients, increased acetylation of histone H3 binding to TNF-α promoters
in diabetic blood monocytes has been found to promote transcription of
this inflammatory gene in a hyperglycemic environment. In contrast,
overexpression of histone deacetylation (HDAC) can reverse this process
[17]. Another study in diabetic mice shows that histone H3K4 methyl­
transferase SET7/9 in monocytes can significantly inhibit
TNF-α-induced inflammation [18]. In a study of diabetic retinopathy,
the levels of H4K20me3 and H3K9ac in the promoter and enhancer of
superoxide dismutase 2 (SOD2) were reported to increase due to hy­
perglycemia, but after the termination of hyperglycemia, their levels did
2
Biomedicine & Pharmacotherapy 156 (2022) 113984
not decrease, and the expression of SOD2 continued to be abnormal
[19].
These studies emphasize the key role of epigenetic histone modifi­
cations in the pathogenesis of diabetes. Along with methylation and
acetylation, other histone modification processes are involved in the
progression of diabetes and complications of diabetes. However, our
knowledge of novel HPTMs in diabetes remains limited due to the lack of
thorough investigation of this relatively new field of study. A strong
understanding of these variations in histone lysine acetylation and
methylation and corresponding regulatory elements may aid in identi­
fying new histone modification markers and significant therapeutic
targets for diabetes.
3. Novel histone modifications in diabetes and complications of
diabetes
3.1. β-hydroxybutyrylation
Due to a rising emphasis on the comprehensive treatment of diabetes,
new dietary replacement strategies have attracted increasing attention
from researchers. The ketogenic diet (KD) is a high-fat, adequate-pro­
tein, and low-carbohydrate diet. As a dietary treatment, KD can reduce
postprandial blood glucose levels, promote insulin sensitivity, reduce
body weight, and even reverse diabetic nephropathy [20–22]. With KD,
the body relies more on burning fat for energy and increases the pro­
duction of ketone bodies, namely β-hydroxybutyrate (BHB), acetoace­
tate, and acetone. KD has primarily been used to increase serum BHB
levels, and it can also be enriched after prolonged exercise, fasting, and
diabetic ketoacidosis [23]. Physiologically elevated BHB levels have
been shown to play a potential protective role in diabetic patients. It has
been reported that BHB may reduce oxidative stress in type 2 diabetic
rats, thereby preventing diabetic cardiomyopathy and diabetic cardiac
death [24]. The underlying epigenetic molecular mechanisms of BHB as
a beneficial factor for diabetes were discovered recently.
In a landmark 2016 study, Kbhb was first identified and character­
ized by Yingming Zhao’s team in a mouse model of streptozotocin (STZ)
induced ketoacidosis using high mass accuracy tandem mass spec­
trometry proteomics. Physiological and pathological changes in BHB
concentration under conditions such as starvation and diabetic ketoa­
cidosis can give rise to many Kbhb histone markers in mouse liver
(Fig. 1). Based on these findings, it is reasonable to assume that the KD
could also cause these changes. Kbhb is mainly enriched in active gene
promoters related to genes up-regulated in the starvation-responsive
metabolic pathway [10]. In addition to indirectly participating in
HPTMs as an HDAC inhibitor [25], BHB can also directly regulate the
complex process of gene transcription as an acyl donor, which greatly
expands the regulatory range of BHB in the field of epigenetics.
Additionally, recent studies have revealed that Kbhb ameliorates
diabetic complications by regulating the expression of certain genes.
Some researchers have pointed out that BHB treatment upregulated the
generation of vascular endothelial growth factor (VEGF) and promoted
endothelial repair after vascular injury in diabetic rats [26]. During this
process, BHB caused a significant increase in total protein Kbhb and
H3K9bhb content in the aorta of diabetic rats. However, whether
H3K9bhb was the initiating factor promoting VEGF gene expression and
which enzymes were involved in this process were not further elabo­
rated in this report. A recent study linked Kbhb to another salutary
biological impact. Luo et al. found that BHB could significantly upre­
gulate the expression of MMP-2 by increasing the content of H3K9bhb in
the MMP-2 promoter in diabetic rats [27], which reduced the patho­
logical changes of glomeruli and decreased the renal fibrosis index in a
concentration-dependent manner. The above research can provide an
essential experimental basis for the clinical application of BHB in
treating diabetic kidney disease (DKD). The expression of VEGF not only
contributes to diabetic angiopathy but also contributes to the damage
repair of diabetic retinopathy and nephropathy. Similarly, MMPs have
D. Li et al. Biomedicine & Pharmacotherapy 156 (2022) 113984

Fig. 1. Sources and regulatory processes of histone malonylation, β-hydroxybutyrylation, and succinylation in diabetes.
Glucose, fatty acids, and amino acids are transported to mitochondria for oxidative decomposition, where they are catalyzed by acyl-COA synthases to generate the
intermediates malonyl-CoA, BHB-CoA, and succinyl-CoA. These three types of acyl-CoA can be transported to the nucleus and catalyzed by the corresponding
acyltransferase or deacylase to regulate Kma, Kbhb, and Ksucc in histone lysine residues. These three novel HPTMs are involved in neural tube defects, vascular
endothelial dysfunction, glomerulosclerosis, and diabetic obesity, respectively. BHB-CoA, β-hydroxybutyryl CoA; TCA cycle, Tricarboxylic acid cycle; SIRT, Sirtuin;
CBP, CREB-binding proteins; GCN5, general control non-depressible 5; NTDs, neural tube defects; DIO, diet-induced obesity.

been found to be involved in the pathogenesis of retinopathy [28–30].
Therefore, it is not clear whether elevated Kbhb levels are tissue-specific
drivers of these genes. Accordingly, we were unable to identify the
maximum beneficial target organs for the up-regulation of VEGF and
MMP2 by Kbhb. The concentration-dependent regulatory role of Kbhb
may need to be further elucidated in a wider range of genes. A recent
proteomics report on BHB quantified the distribution of modification
sites and functional enrichment of Kbhb after BHB treatment in mouse
embryonic fibroblasts. The results showed that BHB-induced Kbhb
proteins were mainly involved in glycolysis/gluconeogenesis, pyruvate
metabolism, and citrate cycle pathways, and 26 % of the Kbhb protein
was located in the nucleus [31]. Single or multiple Kbhb sites in histones
may play an indispensable role in the occurrence and development of
diabetes through their transcriptional regulation of intermediate en­
zymes in glucose metabolism.
3.2. Butyrylation and propionylation
The microbial community can metabolize more than two dozen
short-chain fatty acids (SCFAs), including acetic, propionic, and butyric
acids, which are crucial bacterial fermentation products of high-fiber
foods [32]. Research in recent decades has highlighted the importance
of the composition and abundance of the gut microbiota in the physio­
logical regulation of bacterial hosts, such as inhibiting intestinal in­
flammatory responses and combating oxidative stress [33,34]. The
process of nutrient degradation by the symbiotic microbial community
is also crucial for maintaining the host’s glucose homeostasis. One study
found that butyrate synthesized by gut microbiota can improve glucose
tolerance and reduce glycated hemoglobin A1c levels to alleviate dia­
betes [35,36]. Intake of sodium butyrate can inhibit high
glucose-induced oxidative stress and inflammatory response in mesan­
gial cells, reduce renal fibrosis, and reduce apoptosis to strengthen renal
function in diabetic rats [37]. Propionate also has positive effects on the
function of pancreatic β cells, and in vivo and in vitro analyses show that
it can enhance glucose-stimulated insulin secretion and β-cell mass by
inhibiting apoptosis [38]. The above findings support the crucial regu­
latory role of SCFAs produced in the gut in maintaining glucose ho­
meostasis and attenuating microvascular complications caused by
hyperglycemia. Besides that, microbial signals also calibrate the tran­
scriptional program of host cells through short-chain lysine acylations
directly on target histones.
In 2007, Yue’s group first reported two novel lysine modifications,
Kpr and Kbu [4]. Kbu and Kpr are respectively derived from butyrate
and propionate, and they are catalyzed by ACSS2 to form small active
molecules similar to acetyl-CoA to mediate histone acylation processes
(Fig. 2). Histone Kbu and Kpr may have the possibility of extensive
pathophysiological regulation similar to histone acetylation. Mass
spectrometry reveals they can be catalyzed in vitro by two well-known
acetyltransferases, p300 and CREB-binding proteins (CBP). Kpr and
Kbu are highly dynamic and preferentially enriched in the promoters of
active genes. They can be detected in eukaryotic cells such as yeast cells,
mouse liver, and leukemia cell line U937, reflecting their high conser­
vation [39,40]. It is exciting that these two short-chain lysine acylations
were discovered on histones in the livers of pre-diabetic mice with
high-fat diet-induced obesity (DIO) in 2017 [41]. Specifically, compre­
hensive proteomic analysis of histone samples extracted from known
and unknown histone marks indicated 15 Kpr marks and 12 Kbu marks
among the 45 newly discovered histone lysine acylation marks. The
detection of these histone marks had never been done before in diabetic
animal models. Furthermore, antibody affinity enrichment showed that
Kpr and Kbu marks appeared on histones with a high presence rate.
Recent studies have confirmed that intraperitoneal injection of sodium
butyrate can up-regulate renal histone Pan-butyrylation (Pan-Kbu) and
H3K9 butyrylation (H3K9bu) in DKD mice, and the proinflammatory
factors MCP-1 and IL-6 and the fibrosis indicators COV-IV and TGF-β are
significantly down-regulated. Additionally, the anti-inflammatory and
anti-fibrotic effects of sodium butyrate are reversed after inhibition of
Kbu with A485, a histone modifying enzyme p300 inhibitor [42]. Due to

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D. Li et al. Biomedicine & Pharmacotherapy 156 (2022) 113984

Fig. 2. Sources and regulatory processes of histone propionylation and butyrylation in diabetes.
Microorganisms such as bacteria grown on the surface of intestinal epithelial cells can digest dietary fiber derived from food by fermenting it into more than 20 kinds
of short-chain fatty acids (SCFAs), including propionate and butyrate. These SCFAs can be transported into cells by cell membrane channel monocarboxylate
transporters (MCTs), where they are catalyzed by ACSS2 to generate propionyl-CoA and butyryl-CoA. Meanwhile, the metabolic cycle of mitochondria can also serve
as a secondary source of propionyl-CoA and butyryl-CoA. As histone modification substrates, propionyl-CoA and butyryl-CoA are involved in the occurrence and
development of diabetes, diabetes-related kidney disease, and atherosclerosis under the catalysis of corresponding acyltransferases and deacylases. IR, insulin
resistance; ASCVD, atherosclerotic cardiovascular disease; KAT6, lysine acetyltransferase 6; PCAF, p300/CBP-associated factor; MOZ, monocytic leukemia zinc finger
protein; MOF, male absent on the first; HBO1, histone acetyltransferase binding to ORC1; GCN5, general control non-depressible 5; SIRT, Sirtuin.

the nonspecific nature of enzyme regulation of A485 and the fact that
sodium butyrate is also an important inhibitor of HDAC, this report lacks
experimental results related to the effect of BHB on acetylation and other
histone modifications, and attention should be paid to the dynamic
changes in the abundance of other butyrylation modification sites in the
DKD model. Recent studies have identified a semisynthetic derivative
(LTK-14A) that has inhibitory effects on p300 and specifically inhibits
Kbu without affecting the function of Kac [43]. This finding may
accelerate our investigation of the ambiguous causal relationship be­
tween diabetes and Kbu.
Short-chain histone acylations may provide new insights into the
epigenetic roles of gut microbiota metabolites involved in the patho­
genesis or treatment of diabetes, so further studies need to explore the
specific modification enzymes, modification sites, and effect on the
expression of Kbu and Kpr target genes.
3.3. Succinylation
In most cases, intracellular succinyl-CoA originates from α-ketoglu­
tarate, succinyl-carnitine, and succinate in the mitochondrial tricar­
boxylic acid (TCA) cycle. Moreover, carnitine shuttling during fatty acid
β-oxidation also increases the level of succinyl-CoA [44,45]. The above
two processes are closely related to the glycolipid metabolism abnor­
mality of diabetes. The connection between them reminds us that the
high sugar environment and the metabolic imbalance of aerobic
oxidation of glucose caused by diabetes can increase the level of
succinyl-CoA, which participates in the development of diabetes.
Succinyl-CoA can cross the mitochondrial membrane and produce
Ksucc in cytoplasmic proteins and nuclear histones [46]. In 2011, Zhang
et al. discovered and characterized 69 succinylated lysine sites [5],
opening the door for subsequent functional studies related to histone
modifications. Ksucc is a reversible, dynamic, abundant, and evolu­
tionarily conserved histone mark in eukaryotic cells. It has modification
sites on histones H2A, H2B, H3, and H4. Ksucc may have a significant
effect on chromatin structure and function because it can produce mass
shifts of up to + 100 Da, which is greater than the mass shifts produced
by Kme or Kac. And the addition of succinylation substrates to lysine
residues can cause two charge shifts, which change the positively
charged side chains to negatively charged side chains. These drastic
charge reversals and mass changes can obviously change the binding
ability of histone to DNA, which may be highly correlated with gene
expression regulation.
Current studies on the write-read-erase mechanism of Ksucc found
that SIRT3, SIRT5, and SIRT7 in the deacetylase family have potent
desuccinylase activity [47–49]. Among them, SIRT5 is the most thor­
oughly studied desuccinylase. The levels of succinylation on mito­
chondria, cytoplasm, and histones in the liver cells of SIRT5 gene
knockout mice were significantly increased. Another study found that
overexpression of SIRT5 considerably reduced the levels of apoptosis
factor Caspase-3 and MDA, upregulated the activation of the
anti-apoptosis factor B-cell lymphoma 2 (Bcl-2) in pancreatic β cells
damaged by high blood sugar levels, and improved the insulin secretion
function [50]. These data indicate that SIRT5-mediated Ksucc may be
involved in easing pancreatic β cell apoptosis and promoting the
expression of genes related to insulin secretion, pointing to its potential
role in maintaining glucose homeostasis. In a recent paper, Nie’s
research team reported 30 previously unknown modification sites
identified by HPLC-MS/MS analysis of histone samples extracted from
the liver of a pre-diabetic C57BL/6J mouse model induced by a high-fat
diet for eight weeks. These included nine Ksucc sites, along with Kpr,

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D. Li et al. Biomedicine & Pharmacotherapy 156 (2022) 113984

Kbu, and Kma sites [41], indicating that the three newly discovered
histone acylations in the pre-diabetic mouse model may have some
unrevealed association with diabetes and obesity. Since the procedure in
this study used pan-antibody and did not involve affinity enrichment
analysis for histone modifications of interest, it is reasonable to assume
that these newly discovered histone markers, including histone succi­
nylation, have a high stoichiometric ratio in DIO mouse livers.
Increasing evidence suggests that succinylation is a widespread
protein modification, and it is vital in many diseases, such as cancer,
neonatal cardiac energy metabolism, Alzheimer’s disease [51], and
cardiovascular disease [52]. However, since current studies on the
regulatory mechanism of succinylation generally focus on the enzyme
activity regulating the TCA cycle and the relationship between meta­
bolism and the four aforementioned diseases, little is known about most
other biological functions, even less in the field of diabetes. Therefore, it
is necessary to investigate the lysine succinylation targets of histones in
diabetic models besides H3K79, H4K31, H4K77, H2BK46, and H2BK85
(Table 1) and determine the factors involved in their upstream and
downstream regulation. We require more clear evidence based on new
molecular mechanisms of succinylation and desuccinylation to meet the
needs of future discovery and development of therapies and drugs for
diabetes.
3.4. Malonylation
Malonyl-coA is an essential intermediate in fatty acid synthesis and
β-oxidation, and it is an inhibitor of fatty acid oxidation [53]. Two
distinct isoforms of acetyl-CoA carboxylase (ACC), ACC1 and ACC2, and
malonyl-CoA decarboxylase (MCD) are the key catalytic enzymes of
intracellular lipid metabolism, as they respectively mediate the syn­
thesis and degradation of malonyl-coA [54]. In addition to finding
higher malonyl-CoA concentrations in people with type 2 diabetes, two
studies reported that compared with wild-type mice, higher levels of
glucose and fatty acid oxidation were found in ACC2-/- mutant mice fed
a high-fat and high-carbohydrate diet, and systemic insulin resistance
and hyperglycemia were improved in mice by ACC knockout or use of
ACC2 selective inhibitors [55,56]. Another study found that in mice
with ACC1 and ACC2 double-knockout or phosphorylated inhibition,
malonyl-CoA could not be transformed by acetyl-CoA, resulting in
increased fat production, decreased fatty acid oxidation, increased levels
of insulin resistance, and abnormal glucose tolerance [57]. These results
reveal an intrinsic link between metabolic pathways using malonyl-CoA
as intermediates and the establishment of glucose homeostasis.
Kma is a novel modification based on malonyl-CoA. It was first
identified in 2011 by Peng et al. in Escherichia coli and HeLa cells using
specific malonyl-lysine antibody, high-performance liquid
chromatography-tandem mass spectrometry (HPLC/MS/MS), and pro­
tein sequence-alignment [7]. Functional enrichment analysis showed
that Kma was significantly enriched in glucose and fatty acid metabolic
pathways [58]. Kma has a large molecular size, can cause a mass shift of
+ 86.00039 Da in histones, and is an evolutionally conserved modifi­
cation that is widespread in many species, including humans, fruit flies,
zebrafish, and mice. Therefore, it is reasonable to predict that Kma af­
fects the binding between DNA and histones in many species. With the
deepening of research on Kma, its sites on histones and function in
diabetes gradually surfaced. In the absence of antibody enrichment of
core histones, seven malonylation sites were found in the DIO mouse
model of prediabetes, among which were H3K23, H3K56, H3K79,
H2BK108, H2BK116, and H2BK120 [41]. However, it is not clear which
genes are regulated by modifications at these sites. A recent work
documented that when mouse NE4C cells were cultured with glucose at
different concentrations, the level of Kma was upregulated with the

Table 1
Summary of histone modifications in diabetes.
Histone Models Sites Regulatory Specific functions Ref.
modification enzymes

Kbhb ①Liver from prolonged fasting or H3:K9, K18 Writers: P300/CBP ①it can antagonize aortic endothelial injury by [10],
STZ-induced T1DM ketoacidosis Erasers: SIRT1–3, promoting generation of VEGF; [26],
mouse; HDAC1–3 ②It can upregulate MMP-2 and reduce diabetic [27]
②Kidney from STZ-induced diabetic Readers: not identified glomerular fibrosis.
SD rat
Kma ①Liver from high-fat diet-induced H2A: K74, K75, K95, Writers: Not available It may influence the regulation of genes and [7],
prediabetic mice; K99, K118, K119, K124 Erasers: SIRT2, SIRT3, pathways related to neurological complications [41],
②Brain from neural tube defect H2B:K12, K15, K34, SIRT5 caused by diabetes. [59]
(NTD) embryos of female diabetic K46, K57, K108, K116, Readers: not identified
mice; K120,
③High glucose induced NE4C cell H3:K4, K23, K37, K56,
K79, K119
H4:K5, K8, K12, K16,
K44, K59, K77, K79,
K91
Kpr Liver from high-fat diet-induced H1:K65 Writers: P300/CBP, Not identified [4],[41]
prediabetic mice H2B:K5, K12, K15, GCN5, PCAF, KAT6,
K16, K20, K108, K116 MOF, MOZ, HBO1
H3:K9, K18, K23, K27 Erasers: SIRT1–3, SIRT5
H4:K5, K12, K16 Readers: Bromodomain,
YEATS
Kbu ①Liver from high-fat diet-induced H2A:K9 Writers: P300/CBP, ①It may promote the transcriptional expression of [4],
prediabetic mice; H2B:K5, K12, K15, GCN5, PCAF, KAT6, Ins1 and Ins2 genes; [41],
②Kidney from DKD mice induced by K16, K20, H3:K18, K27 MOF, ②It may decrease the transcriptional expression of [42]
high-fat diet and STZ; H3:K9 HBO1 MCP-1, IL-6, TGF-β genes;
③High glucose induced SV- H4:K5, K8 Erasers: SIRT1–3
40MES13 cell Readers: Bromodomain,
YEATS
Ksucc Liver from high-fat diet-induced H1:K171 Writers: P300/CBP, Not identified [5],
prediabetic mice H2B:K46, K85, K108, GCN5 [41],
K116, K120 Erasers: SIRT3, SIRT5, [45]
H3: K23, K27, K56, SIRT7
K79, K122 Readers: YEATS
H4: K31, K77, K79, (GAS41)
K91

5
D. Li et al.
increase of glucose intervention concentration [59], suggesting a posi­
tive correlation between glucose concentration and the level of histone
malonylation, which may be related to diabetic neuropathy mediated by
high glucose. Neural tube defects (NTDs) are common congenital mal­
formations in the fetuses of diabetic women. Western blotting has
detected increased histone malonylation levels of embryos with NTDs in
STZ-induced female diabetic mice. These findings remind us that histone
malonylation may also participate in neurological complications of
diabetes, and the enzymes regulating malonylation may be suitable
targets for treating such complications.
SIRT5 is the primary demalonylase and it can mildly reduce Kma.
There are only a few known modification sites of Kma, such as H3K4,
H3K23, H3K56, H4K8, and H4K77. Key future directions in the study of
Kma should be to identify the many other “write-erase” enzymes
mediating this modification, determine its specific or nonspecific sites,
determine the levels of Kma in critical locations on histones associated
with diabetes, and assess the role of Kma in diabetes.
3.5. Benzoylation
Kbz is a novel histone modification added to lysine residues by the
food additive sodium benzoate and endogenous intestinal metabolite
benzoyl-CoA as substrates. It was discovered in 2018 as a functionally
conserved modification that exists in Drosophila, mouse, and human cell
lines. Because Kbz is the only known type of HPTM with an aromatic
ring structure and its mass migration can be as high as + 104 Da, the
probability of it regulating neighboring genes is greatly increased.
Notably, intracellular benzoyl-CoA and global Kbz levels significantly
increased even when sodium benzoate was ingested at concentrations
below the maximum allowable addition concentration (~0.1 %) in foods
set by the US Food and Drug Administration (FDA). KEGG functional
enrichment analysis hinted that Kbz might regulate insulin secretion
[11]. However, no further experiments in vivo or in vitro have been
conducted to verify this possible relationship. Due to the unusual
structure and prevalence of sodium benzoate in the food industry, it will
be fascinating to further investigate the role of Kbz in the regulation of
genes related to insulin secretion, which could contribute to our un­
derstanding of the pathological process of diabetes and the development
of dietary strategies.
4. Conclusions and prospects
With the advancement of proteomic technology, a mounting number
of histone acylation reactions have been discovered and confirmed to be
involved in the progress of diabetes and associated complications.
Indeed, almost all of the novel histone modifications discussed in this
review are highly enriched in the histones of diabetic models. To some
extent, they also reveal the common pathologic mechanisms between
diabetes, obesity, and other metabolic diseases. Nevertheless, the re­
lationships between novel histone modifications and diabetes are still
understood on only a superficial level, and the associated molecular
mechanisms have not been thoroughly studied. Because only a small
amount of evidence suggests that these novel HPTMs may be beneficial
for part of diabetes complications, it is not clear whether novel HPTMs
have pathological effects on diabetic metabolic homeostasis in terms of
genetics and pharmacology, so more extensive and in-depth patho­
physiological studies are urgently needed.
Since most HPTMs are dynamic and reversible, intervention of up­
stream or downstream targets in the formation of these modifications
could give rise to new diabetes treatments. Moreover, these HPTMs
share the same acyl-CoA synthetases, acyltransferases, and deacylases,
which can be taken advantage of to quickly find their regulatory en­
zymes and corresponding substrates and conduct in-depth research on
their biological functions. For example, we can continue to explore
whether the classic acyltransferase P300/CBP can also catalyze the
addition of novel histone modifications identified in the future. ACSS2 is
6
Biomedicine & Pharmacotherapy 156 (2022) 113984
one of the known acetyl-CoA synthetases involved in important meta­
bolic processes in the body. A future research direction should be to
verify whether ACSS2 can affect the synthesis of other types of acetyl-
CoA, and to investigate how to make ACSS2 selectively regulate one
novel histone modification without affecting its other functions. Some
studies have found that acyl-CoA synthetases, acyltransferases, and
deacylases are involved in the pathogenesis of diabetes complications,
but the regulation they mediate does not target specific modifications.
Therefore, while studying one of these novel HPTMs, it is necessary to
detect the dynamic changes of other modifications such as histone
acetylation. We still need to discover and utilize specific regulatory
enzymes to determine whether novel HPTMs could be beneficial for
diabetes control. A further complication is that we do not yet know
whether different HPTMs cooperate with or antagonize each other.
Therefore, the functions and underlying mechanisms of two or more
novel combinations of HPTMs need to be investigated in the same
experiment. Moreover, we should continue to investigate whether, like
Kme and Kac, other novel HPTMs found in diabetes and complications of
diabetes mediate the persistent epigenetic change called "metabolic
memory" that may be transferred to offspring.
Above all, this review discusses how some novel HPTMs—repre­
sented by β-hydroxybutyrylation, malonylation, propionylation, butyr­
ylation, and succinylation—are closely or potentially related to the
pathophysiology of diabetes and complications of diabetes. With the
development of new site-specific antibodies in the future, the accuracy
and detection range of modified omics technology will see great im­
provements. The catalog of histone modifications will continue to
expand, and future work will need to focus on establishing a complete
map of the associations between histone modification and diabetes and
exploring the precise functions and mechanisms of these associations.
Such work will enable the development of new prevention and treat­
ment targets for diabetes and its complications.
Funding
This work was supported by the National Natural Science Foundation
of China (NO.81970676) and the Luzhou-Southwest Medical University
cooperation project (NO.2021LZXNYD-P02).
Author contributions
All authors conceived the manuscript structure and contributed to
the writing and editing.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Data availability
Data will be made available on request.
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