Next Article in Journal
Effect of Plyometric Exercises of Lower Limb on Strength, Postural Control, and Risk of Falling in Stroke Patients
Previous Article in Journal
Drug Utilization of Rifaximin-α in Patients with Hepatic Encephalopathy: Evidence from Real Clinical Practice in Italy
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Medicina
Volume 61
Issue 2
10.3390/medicina61020222
medicina-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Review

Novel Insight of N6-Methyladenosine in Cardiovascular System

by
Huan Zhang
1,†,
Wei Lu
1,†,
Haoyue Tang
1,
Aiqun Chen
1,
Xiaofei Gao
1,
Congfei Zhu
2,* and
Junjie Zhang
1,*
1
Department of Cardiology, Nanjing First Hospital, Nanjing Medical University, Nanjing 210006, China
2
Department of Cardiology, Lianshui County People’s Hospital, Affiliated Hospital of Kangda College, Nanjing Medical University, Huaian 223400, China
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Medicina 2025, 61(2), 222; https://doi.org/10.3390/medicina61020222
Submission received: 28 November 2024 / Revised: 19 January 2025 / Accepted: 24 January 2025 / Published: 26 January 2025
(This article belongs to the Section Cardiology)
Browse Figures
Review Reports Versions Notes

Abstract

:
N6-methyladenosine (m6A) is the most common and abundant internal co-transcriptional modification in eukaryotic RNAs. This modification is catalyzed by m6A methyltransferases, known as “writers”, including METTL3/14 and WTAP, and removed by demethylases, or “erasers”, such as FTO and ALKBH5. It is recognized by m6A-binding proteins, or “readers”, such as YTHDF1/2/3, YTHDC1/2, IGF2BP1/2/3, and HNRNPA2B1. Cardiovascular diseases (CVDs) are the leading cause of morbidity and mortality worldwide. Recent studies indicate that m6A RNA modification plays a critical role in both the physiological and pathological processes involved in the initiation and progression of CVDs. In this review, we will explore how m6A RNA methylation impacts both the normal and disease states of the cardiovascular system. Our focus will be on recent advancements in understanding the biological functions, molecular mechanisms, and regulatory factors of m6A RNA methylation, along with its downstream target genes in various CVDs, such as atherosclerosis, ischemic diseases, metabolic disorders, and heart failure. We propose that the m6A RNA methylation pathway holds promise as a potential therapeutic target in cardiovascular disease.

1. Introduction

More than 140 types of chemical modifications have been discovered in RNA. Among these modifications, RNA methylation involves the addition of methyl groups to specific nucleotide residues in RNA, such as N1-methyladenosine (m1A), N6-methyladenosine (m6A), N6,2′-O-dimethyladenosine (m6Am), and 2′-O′-methylation (2′-OMe) [1,2,3]. Among these modifications, the methylation of mRNA at the N-6th position of the adenosine residue (m6A) has been extensively studied since its discovery in the 1970s. It is the most common methylation modification found in the internal sequence of mRNA in eukaryotes, and it is the most abundant and evolutionarily conserved reversible post-transcriptional modification [4,5,6]. Functionally, m6A is involved in various aspects of RNA biology, including RNA stability, nuclear export, translation efficiency, and mRNA degradation [2,3,7]. In addition to regulating RNA coding sequences (mRNA), m6A methylation also plays a regulatory role in non-coding RNAs such as microRNAs, long non-coding RNAs, and circular RNAs, thereby influencing gene expression in diverse physiological processes [8,9,10].
Cardiovascular diseases (CVDs) are the leading cause of morbidity and mortality worldwide, and their pathological mechanisms are complex [11]. Despite significant advances in the diagnosis, treatment, and prognosis of CVDs, there is an urgent need for new diagnostic biomarkers and treatments to reduce mortality and improve therapeutic outcomes for patients with CVDs.

2. m6A RNA Methylation

The rapid advancement of bioinformatic analyses and high-throughput sequencing technologies has revealed that m6A modification primarily occurs at the consensus motif RR (m6A) CH (R = A or G, H = A, C, or U) in long internal exons, near stop codons, or in the 3′ untranslated region (UTR) [7,12] (Figure 1). Importantly, m6A modification is dynamic and its methylation levels undergo dynamic changes, regulated by various methyltransferases (“writers”), m6A-binding proteins (“readers”), and demethylases (“erasers”) [13,14]. These three groups of proteins are responsible for the addition, removal, and recognition of m6A RNA modification.
Studies have shown that m6A modification can alter RNA secondary structures or be recognized by “readers” to regulate the metabolism of methylated mRNAs, thereby exerting various functions [15]. The process of RNA modification by m6A begins during transcription. The addition and removal of m6A mostly occur in the nucleus, where specific nuclear “readers” can bind to m6A and potentially affect mRNA splicing and other nuclear processes. After export to the cytoplasm, m6A interacts with specific cytoplasmic reader proteins that influence RNA stability, translation efficiency, and localization [2] (Table 1).
M6A “writers” consist of METTL3/14, KIAA1429, WTAP, zinc finger CCCH domain-containing protein 13 (ZC3H13), and RBM15/15B [31,32]. Among these, METTL3 and METTL14 are considered the most critical core components of the m6A methyltransferase complex [16]. METTL3 acts as the catalytic subunit, while METTL14 plays a structural role and activates METTL3 through allosteric and RNA substrate recognition [16,33]. WTAP interacts with METTL3-METTL14 and is the third subunit of the complex [17,18]. RBM15/15B is responsible for recruiting the methylation complex to XIST [19]. ZC3H13 plays a critical role in anchoring WTAP, Virilizer, and Hakai in the nucleus for m6A methylation regulation and mESC self-renewal [20]. KIAA1429, as a top interactor with WTAP, recruits specific cleavage and polyadenylation specificity factors, leading to a longer 3′UTR selection [21]. ZC3H13, KIAA1429, and RBM15/RBM15B play key roles in localizing the methyltransferase complex in nuclear speckles and U-rich regions adjacent to m6A sites in mRNAs.
FTO (FAT mass and obesity-associated protein) and ALKBH5 (ALKB homolog 5 protein) belong to the ferric divalent/α-ketoglutarate dependent dioxygenase ALKB protein family and exhibit efficient oxidative demethylation activity targeting N6-methyladenosine residues in RNA [2,23]. Both FTO and ALKBH5 are located in the nuclear speckle area, bind to m6A-modified RNA, catalyze the oxidation of m6A to adenine, and produce the intermediate products hm6A (N6 hydroxymethyl adenosine) and further oxidation product f6A (N6 formyl adenosine).
“Readers” are proteins that recognize m6A methylation sites on RNAs, bind to mRNA, and influence nuclear export, RNA stability, or degradation. The YTH domain-containing proteins, including YTHDC1, YTHDC2 (DC2), YTHDF1 (DF1), DF2, and DF3, were the first m6A-binding proteins to be discovered [14]. YTHDF1 promotes mRNA translation, while YTHDF2 accelerates the decay of m6A-modified transcripts. The role of YTHDF3 is complex, as it facilitates YTHDF1’s role in promoting translation, and knockdown of YTHDF3 results in reduced translation efficiency of mRNA targets of both YTHDF3 and YTHDF1 [24,25]. A recent study showed that YTHDC1 blocks SRSF10 mRNA binding by selectively recruiting the SRSF3 mRNA pre-splicing factor, thereby promoting exon inclusion in the target transcript [34]. Nuclear export of m6A-methylated transcripts may be facilitated by YTHDC1 through interaction with nuclear transport receptors [27]. YTHDC2 is an N6-methyladenosine-binding protein that regulates mammalian spermatogenesis by enhancing the translation efficiency of its targets and also decreasing their mRNA abundance [28]. IGFBP proteins, including IGF2BP1/2/3, can regulate RNA localization, translation, and stability [29]. The heterogeneous nuclear ribonucleoprotein (hnRNP) family, including hnRNPG, hnRNPC, and hnRNPA2B1, is another group of m6A “reader” proteins. Wu et al. found that m6A can enhance the ability of hnRNPA2/B1 to enhance nuclear events, such as pri-miRNA processing [30].

3. Role of m6A in Cardiovascular Disease

3.1. Risk Factors Associated with CVDs

Unhealthy diet, obesity, and diabetes are well-recognized risk factors involved in the pathophysiology of numerous chronic cardiovascular diseases (CVDs). In this comprehensive review, we aim to summarize the current state of research regarding the mechanisms underlying m6A methylation and its role in the development of these CVD risk factors. By examining the existing literature, we can gain insights into the molecular processes driving m6A methylation and its contribution to the pathogenesis of CVDs. Importantly, these findings offer valuable information that can guide future therapeutic interventions and help identify potential targets for the prevention and management of CVDs. Table 2 provides a summary of the key findings and future therapeutic prospects in this field.

3.1.1. Glucose Metabolism

The regulation of glucose metabolism is crucial in maintaining cardiovascular health, as diabetes or hyperglycemia can have detrimental effects on the cardiovascular system. Correcting or preventing abnormalities in glucose metabolism can significantly reduce the incidence of cardiovascular disease. Yang et al. demonstrated that high glucose stimulation enhanced the expression of FTO, which resulted in decreased m6A methylation. FTO then triggered the mRNA expression of FOXO1 and G6PC, which are associated with gluconeogenesis [35]. This finding suggests that controlling blood glucose levels by regulating FTO expression could be a potential method for managing glucose metabolism.
Additionally, FTO plays a regulatory role in improving glucose levels and may have implications in protecting the injured heart or improving heart function in heart failure patients. In a recent study, a team created a model of transverse aortic constriction-induced heart failure (HF) in mice and found that cardiac fibrosis and hypertrophy were ameliorated in mice overexpressing FTO. They further observed that 18F-FDG uptake, an indicator of glucose uptake, was significantly increased in mice overexpressing FTO, confirming FTO’s role in regulating glucose uptake and glycolysis for the attenuation of cardiac dysfunction [41].
The depletion of m6A levels in pancreatic β-cells was found to induce cell-cycle arrest and impair insulin secretion by decreasing AKT phosphorylation and PDX1 protein levels, as reported by De Jesus et al. [36]. Another study by Krüger et al. in 2019 supported this concept, demonstrating that the loss of endothelial FTO protected mice from high-fat-diet-induced glucose intolerance and insulin resistance by increasing AKT phosphorylation in endothelial cells and skeletal muscle [37].

3.1.2. Adipogenesis and Obesity

The deposition of lipids in the arterial intima and the formation of foam cells are closely associated with adipogenesis. Obesity, being a major risk factor for coronary heart disease, necessitates effective control of adipogenesis to prevent atherosclerosis and subsequent coronary syndromes such as myocardial infarction.
Current evidence strongly supports the role of FTO in promoting adipogenesis. KLF4, an essential early regulator of adipogenesis, is normally suppressed by C/EBPβ through a negative feedback loop involving Krox20 and KLF4 expression [42]. Wu et al. found that FTO depletion in porcine and mouse preadipocytes inhibited adipogenesis through the JAK2-STAT3-C/EBPβ signaling pathway [38]. Zhao et al. revealed that FTO controls the exonic splicing of RUNX1T1, an adipogenesis-related transcription factor, by regulating m6A levels around splice sites, thereby promoting adipogenesis [39]. The relationship between FTO and autophagy remains controversial. Wang et al. demonstrated that FTO knockdown inhibited autophagy and adipogenesis by decreasing the expression of ATG5 and ATG7, providing a new mechanism to inhibit adipogenesis [40]. Furthermore, in a study conducted by Song et al., it was verified that Zinc finger protein 217 (Zfp217), a known oncogenic protein, promotes adipogenesis through m6A mRNA methylation via FTO and YTHDF2. Interestingly, the study also highlighted that YTHDF2 blocks the demethylase activity of FTO, leading to increased m6A levels, while Zfp217 functions as a regulator to maintain FTO’s m6A demethylation activity [43]. These findings suggest that FTO knockdown or YTHDF2 overexpression could serve as potential targets for obesity therapy aimed at the prevention of cardiovascular diseases.

3.2. Function of m6A in CVDs

Currently, numerous studies have investigated the role of m6A in cardiovascular diseases (CVDs), including ischemia–hypoxia injury, atherosclerosis, acute myocardial infarction (AMI), and heart failure (Table 3, Figure 2). The effects of m6A modification vary across different CVDs. In this review, we aim to explore the relationship between m6A and CVDs in order to identify potential promising therapeutic targets.

3.2.1. m6A and Ischemia–Hypoxia Injury

Ischemia and hypoxia are recognized as common causes of vascular injury. TFEB is a master regulator of lysosomal biogenesis and autophagy genes. In cardiomyocytes subjected to hypoxia–reoxygenation, the increased expression of METTL3 regulates the m6A modification of TFEB mRNA by promoting HNRNPD association with TFEB pre-mRNA [46]. This dysregulation impairs autophagic flux and enhances apoptosis in hypoxia–reoxygenation-treated cardiomyocytes. Conversely, ALKBH5 reverses the hypoxia–reoxygenation-mediated m6A modification of TFEB mRNA in cardiomyocytes, providing a potential mechanism for cellular prognosis regulation by METTL3 through autophagy modulation under ischemic and hypoxic conditions. Apart from TFEB, METTL3 has also been shown to promote DGCR8 binding to pri-miR-143-3p, thereby enhancing miR-143-3p expression and inhibiting the transcription of PRKCE, a gene implicated in autophagy, which further exacerbates cardiomyocyte pyroptosis and myocardial ischemia–reperfusion injury [69].
Interestingly, there is evidence suggesting contrasting roles of METTL3 and ALKBH5 in response to ischemia–hypoxia injury. While METTL3 exacerbates injury to cells under ischemic–hypoxic conditions, ALKBH5 contributes to angiogenesis maintenance in endothelial cells following acute ischemic stress by reducing SPHK1 m6A methylation and downstream eNOS-AKT signaling [47]. In addition, ALKBH5 overexpression in cardiomyocytes leads to increased translation of YAP, which undergoes YTHDF1-mediated m6A modification, enhancing the proliferative capacity of human and mouse cardiomyocytes [48]. However, it appears that ALKBH5 may sometimes hinder angiogenesis. Zhao et al. demonstrated that ALKBH5 overexpression attenuates blood flow recovery and angiogenesis post-ischemic injury by promoting WNT5A mRNA decay and decreasing its half-life [49]. These contradictory findings suggest that ALKBH5 may have different roles in distinct stages of disease progression or during different periods of human development. Further investigation is needed to elucidate the role of m6A in cardiovascular diseases, particularly in vascular injury.
ULK1, serving as a protein kinase activated upon autophagy stimulation, plays a critical role in recruiting other autophagy-related proteins to the autophagosome formation site [70]. Jin et al. demonstrated in a 2018 study that FTO demethylates ULK1 transcripts, thereby prolonging the half-life of ULK1 transcripts and promoting autophagy while inhibiting cellular apoptosis [71]. Similarly, FTO overexpression inhibits apoptosis in hypoxia–reoxygenation-treated myocardial cells through the regulation of m6A modification in Mhrt [50]. Additionally, Tao Yin et al. found that exosome-based WTAP siRNA delivery ameliorated myocardial ischemia–reperfusion injury by influencing the m6A modification of TXNIP mRNA [51].

3.2.2. m6A and Atherosclerosis

m6A in Vascular Smooth Muscle Cell (VSMC) Differentiation and Angiogenesis

VSMC differentiation is a common phenotype observed in various cardiac vascular diseases, particularly in atherosclerosis [72]. One study investigated the promotion of the differentiation of adipose-derived stem cells (ADSC) into VSMCs under hypoxic stress. The expression pattern of METTL3 in their experiment was consistent with that of VSMC-specific markers, such as α-SMA, SM22α, and calponin, suggesting an important role of METTL3 in VSMC differentiation and potential changes in VSMCs in CVDs [73]. Additionally, circYTHDC2, a non-coding endogenous RNA, has been shown to promote the dedifferentiation of VSMCs into a “synthetic type” through the regulation of TET2 expression. The m6A modification mediated by YTHDC2 stabilizes circYTHDC2, highlighting the potential of the YTHDC2/circYTHDC2/TET2 pathway as an important target to inhibit atherosclerosis through dedifferentiation methods [74].
Apart from its role in differentiation, m6A can also mediate the process of angiogenesis. Parial et al. observed that METTL3 activates the phosphorylation of PHLPP2-mTOR-AKT signaling to enhance angiogenesis [75]. Furthermore, METTL3 enhances the translation of LRP6 and DVL1 in a YTHDF1-dependent manner, thereby regulating the Wnt signaling pathway to exert its angiogenic role [76]. These studies suggest that targeting m6A modification holds promise as a strategy for the treatment of angiogenic diseases.

m6A and Calcification

Calcification is frequently observed in atherosclerosis and valvular diseases, often leading to plaque rupture, thrombosis, valve sclerosis, and valve insufficiency, which can result in serious complications like heart failure. Zhou et al. reported that METTL3 is highly expressed in human calcified aortic valves compared to normal valves. Mechanistically, METTL3 promotes the osteogenic differentiation of human aortic valve interstitial cells by suppressing TWIST1 expression in an m6A-YTHDF2-dependent manner [44]. Additionally, Chen et al. demonstrated that decreasing METTL14 expression in calcified arteries attenuates indoxyl sulfate-induced m6A modification and reduces human artery smooth muscle cell (HASMC) calcification. Knockdown of METTL14 may enhance vascular repair function by reducing calcification, presenting a potential therapeutic approach for atherosclerosis and valvular diseases [45].

m6A in Atherosclerosis

Atherosclerosis, the leading cause of CVD, has a high mortality rate among the affected population. The involvement of inflammation in the pathogenesis of atherosclerosis and its complications has gained considerable attention [77]. Chien et al. discovered that oscillatory stress (OS) up-regulates METTL3 expression, leading to increased NF-κB p65 Ser536 phosphorylation and enhanced monocyte adhesion, thus promoting atherosclerosis. Mechanistically, METTL3-mediated hypermethylation stabilizes NLRP1 mRNA while inducing the degradation of KLF4 mRNA under OS. The m6A hypermethylation is recognized by YTHDF1 and YTHDF2 reader proteins, respectively [52]. Another study investigating the effect of m6A methylation on endothelial cells showed that METTL3 is highly expressed in oxidized low-density lipoprotein (ox-LDL)-induced dysregulated human umbilical vein endothelial cells (HUVECs). METTL3 knockdown prevents the progression of atherosclerosis by inhibiting the JAK2/STAT3 pathway via IGF2BP1 [54]. These findings provide a basis for targeting METTL3 molecules and their downstream pathways to delay the progression of atherosclerosis.
Inflammation increases blood monocyte adhesion and migration into the subendothelial space, which is a critical event in the development of atherosclerosis [78]. METTL14, induced by TNF-α, promotes FOXO1 expression, leading to the increased transcription of VCAM-1 and ICAM-1 by enhancing m6A modification and inducing the endothelial cell inflammatory response and atherosclerotic plaque formation [53]. Moreover, METTL14 promotes DGCR8-mediated processing of pri-miR-19a, leading to the formation of mature miR-19a in atherosclerotic vascular endothelial cells (ASVECs), thereby promoting the proliferation and invasion of ASVECs [55]. Therefore, METTL14 plays a major role in endothelial cell activation and represents a novel therapeutic target.
The role of VSMCs in atherosclerosis has been extensively studied, highlighting their capability to switch to transitional, multipotential cells adopting various cellular states, including inflammation, ossification, and collagen matrix deposition [79]. Recently, it has been shown that m6A modification is involved in the regulation of VSMC behavior. For instance, dihydroartemisinin alleviates angiotensin II-induced VSMC proliferation and inflammatory response by blocking the FTO/NR4A3 axis [56]. TPNS inhibits VSMC proliferation, migration, and intimal hyperplasia by regulating the WTAP/p16 signaling pathway [57].

3.2.3. m6A and Acute Myocardial Infarction

Coronary atherosclerosis is a chronic disease characterized by stable and unstable periods. During unstable periods, when inflammation is activated in the vascular wall, patients may experience myocardial infarction (MI). Inflammation plays a fundamental role in atherogenesis and the pathophysiology of ischemic events [80].
M1 macrophages exhibit robust antimicrobial and antitumoral activity, but they also mediate tissue damage induced by reactive oxygen species (ROS), impairing tissue regeneration and wound healing [81]. Liu et al. suggested that METTL3 directly methylates STAT1, the master transcription factor controlling M1 macrophage polarization [82]. The inflammatory pathways mediated by METTL3 are not limited to a single mechanism. In Wang’s study, Mettl3-mediated m6A modification of CD40, CD80, and TLR4 signaling adaptor Tirap transcripts enhanced their translation in dendritic cells, strengthening TLR4/NF-κB signaling-induced cytokine production [83]. Therefore, METTL3 can serve as an anti-inflammatory target during myocardial infarction to reduce tissue damage. Regarding tissue recovery after an ischemic event, METTL3 knockdown may be a feasible approach. Loss of METTL3/m6A impairs the maturation of pri-miR-143, resulting in low expression of miR-143-3p, which in turn leads to high expression of Yap and Ctnnd1. This promotes cardiomyocyte proliferation and endogenous heart regeneration after MI [58].
We must not overlook the tissue damage caused by ischemia–reperfusion (I/R) injury, which is a significant cause of death in MI. Such damage occurs when blood supply is restored, and excessive free radicals attack the tissues. Chen’s study showed a positive correlation between the expression of ALKBH5 in the infarct area and the levels of CK-MB and LDH in peripheral blood. Additionally, ALKBH5 was found to regulate the content of amino acids involved in the tricarboxylic acid (TCA) cycle and mediate the activity of key enzymes, thereby affecting cell metabolism and survival [59]. Moreover, ALKBH5 has been shown to activate the EGFR-PI3K-AKT-mTOR signaling pathway, enhancing the stability of BCL-2 mRNA and promoting the interaction between Bcl-2 and Beclin1, thereby inhibiting autophagy [84]. Kun Yang et al. demonstrated that ALKBH5 promotes the stability of ErbB4 mRNA and the degradation of ST14 mRNA through m6A demethylation, leading to improved fibroblast-to-myofibroblast transformation and better post-MI repair [60]. Considering the role of ALKBH5 in MI, inhibiting ALKBH5 could be a potential method to improve heart dysfunction after MI.
m6A is also involved in tissue repair after ischemic events. The reduced expression of METTL14 in human artery smooth muscle cells (HASMCs) decreases calcification and enhances vascular repair function [45]. However, Pang et al. found that METTL14 attenuates cardiac ischemia–reperfusion injury by increasing the m6A modification of Wnt1 mRNA, leading to up-regulation of the Wnt1 protein and subsequent activation of the Wnt1/β-catenin signaling pathway [61]. In fact, N6-methyladenosine and the Wnt protein family have close interactions. In a recent study in 2021, ALKBH5 was found to decrease the stability of WNT5A, thereby impeding angiogenesis in hypoxic cardiac microvascular endothelial cells [49].

3.2.4. m6A and Heart Failure (HF)

Heart failure occurs as a consequence of abnormalities in cardiac structure, function, rhythm, or conduction [85]. Despite improvements in survival rates, the absolute mortality rates for HF remain around 50% within 5 years of diagnosis [86]. Researchers have begun exploring the potential of m6A-mediated approaches for improving heart failure. Zhang et al. found that the expression of METTL3, METTL4, KIAA1429, FTO, and YTHDF2 was significantly up-regulated in patients with heart failure with preserved ejection fraction (HFpEF) compared to healthy controls, suggesting that m6A methylation may be a therapeutic target for HFpEF interventions [87].
In addition to restoring contractile protein expression, such as SERCA2, which can alleviate cardiac insufficiency, Mathiyalagan et al. provided experimental evidence that FTO overexpression decreased cardiac fibrosis and enhanced angiogenesis in the ischemic myocardium [62]. This conclusion aligns with the results of Berulova et al., who analyzed m6A RNA methylation through next-generation sequencing and found alterations in the m6A landscape in heart hypertrophy and heart failure [88]. Furthermore, they demonstrated that cardiomyocyte-specific FTO knockdown mice exhibited impaired cardiac function compared to control mice. Hence, the role of FTO is closely associated with recovering cardiac function. In a recent 2024 study, macrophage-specific knockout of ALKBH5 inhibited angiotensin II-induced macrophage-to-myofibroblast transition, subsequently ameliorating cardiac fibrosis and dysfunction. This mechanism involves the targeting of interleukin-11 (IL-11), leading to increased IL-11 mRNA stability and protein levels [65].
Contrary to demethylation, although METTL3 is positively related to angiogenesis [75,76], it serves as a negative regulator in HF. Hinger et al. found increased levels of m6A and METTL3 in human non-ischemic failing hearts [89]. In a recent study, silencing METTL3 reduced m6A modification levels in fibrosis-related genes and decreased myocardial fibrosis in mice with myocardial infarction. This was achieved by inhibiting proliferation, fibroblast-to-myofibroblast transition, and collagen accumulation [90]. Maslinic acid has been demonstrated to protect against pressure-overload-induced cardiac hypertrophy by blocking METTL3-mediated m6A methylation [91]. A study by Dorn et al. identified enhanced METTL3-mediated methylation of mRNA on N6-adenosines in response to hypertrophic stimuli, driving cardiomyocyte hypertrophy in vitro and in vivo [92]. However, Kmietczyk’s experiments with METTL3-overexpressing mice and control mice subjected to transverse aortic constriction surgery to induce pathological hypertrophy led to a contrasting finding—pathological hypertrophic cellular growth was attenuated in the hearts of METTL3-overexpressing mice [93]. Kmietczyk explained this contradiction by suggesting that Dorn et al. used a transgenic model in the FVB background, while the in vivo studies utilized a C57Bl6/N background, and various methods were employed to overexpress METTL3.
In Xu et al.’s study, YTHDF2 was also found to increase in human HF samples. Moreover, YTHDF2 protein levels increased in cell and animal models of cardiac hypertrophy stimulated by isoproterenol or phenylephrine. Mechanistically, YTHDF2 recognizes the m6A site on Myh7 (beta-myosin heavy chain) mRNA, promoting its degradation and thereby alleviating cardiac hypertrophy [63]. MiR-133a represses cardiac hypertrophy and hypoxia-induced apoptosis [94,95]. Qian et al. found that IGF2BP2 binding to the m6A-modified site could enhance the accumulation of the miR-133a-AGO2-RISC complex on its targets, augmenting the repression effect of miR-133a and ultimately inhibiting cardiac hypertrophy [64].

3.2.5. m6A and Other CVDs

The FTO gene is strongly expressed in the hypothalamus, a brain structure involved in blood pressure regulation. Marcadenti et al. demonstrated that common genetic variants of FTO rs9939609 are negatively associated with diastolic and mean blood pressure in men with hypertension [96]. Through large-scale genome-wide association studies, Mo et al. recently revealed the crucial role of m6A in blood pressure regulation. They identified 1236 m6A-SNPs, such as rs9847953 and rs197922, that are potentially associated with blood pressure levels [97]. However, the underlying mechanism of this association still needs further exploration.
In a study on hypoxia-mediated pulmonary hypertension (HPH), researchers identified a transcriptome-wide map of m6A circRNAs and found a reduction in m6A levels in circRNAs in lungs exposed to hypoxia. Furthermore, m6A influenced the circRNA-miRNA-mRNA co-expression network in hypoxia, involving circXpo6 and circTmtc3, indicating the important role of m6A in HPH [98]. In a recent study using a rat model exposed to hypoxic conditions, increased YTHDF2 recognition of METTL3-mediated m6A modification in PTEN mRNA promoted PTEN degradation via PI3K/Akt signaling, leading to the excessive proliferation of pulmonary artery smooth muscle cells (PASMCs) [66]. Additionally, YTHDF1 was found to promote pulmonary hypertension by increasing the translational efficiency of MAGED1, which is involved in the hypoxia-induced proliferation of PASMCs through up-regulating PCNA [67]. These pieces of evidence suggest that down-regulating the level of N6-methyladenosine could be useful in the treatment of HPH.
In the context of aneurysms, similar to how METTL14 promotes DGCR8-mediated processing of pri-miR-19a in atherosclerosis [69], METTL3-dependent m6A methylation promotes primary miR-34a maturation through DGCR8, leading to decreased SIRT1 expression and an aggravated aneurysm formation [68]. This pathway represents a novel therapeutic target and diagnostic biomarker for aneurysm treatment. Vascular smooth muscle FTO promotes aortic dissecting aneurysms via m6A modification of Klf5, enhancing the GSK3β signaling pathway [99]. Moreover, Tan Li et al.’s genome-wide approaches revealed that aneurysm-associated m6A-SNPs might be linked to aneurysm pathogenesis by influencing local gene expression through m6A modification. They specifically identified two m6A-SNPs, NECTIN2 rs6859 and HPCAL1 rs10198139 [100].

3.2.6. m6A in CVDs in the Era of Artificial Intelligence and Machine Learning

Contemporary methodologies that incorporate AI technology and chemosynthesis in drug discovery offer benefits including increased speed, user friendliness, and cost efficiency [101]. They applied a high-throughput fluorescence polarization assay to screen selective inhibitors of FTO from an older drug library containing 900 drugs and finally identified meclofenamic acid as an inhibitor of FTO.
As for in the field of CVDs, a multi-ethnic study of atherosclerosis represented that machine learning in conjunction with deep phenotyping improves prediction accuracy in cardiovascular event prediction in an initially asymptomatic population, and these methods may lead to greater insights into subclinical disease markers without a priori assumptions of causality [102]. In this work, Bharath et al. confirmed the influence of certain markers and risk factors on CV events, such as TNF-α SR and IL2 SR. Such an encouraging study made people believe that machine learning may be useful to characterize cardiovascular risk, predict outcomes, and identify biomarkers.
At present, some teams have begun to combine m6A with artificial intelligence and machine learning to predict and diagnose the occurrence and development of CVDs. In 2021, Xu-shen Xiong et al. tried to excavate genetic drivers of m6A methylation in human tissues, including the heart and muscle [103]. First, they reported 129 transcriptome-wide m6A profiles, covering 91 individuals and 4 tissues (brain, lung, muscle, and heart) from GTEx/eGTEx. And they integrated m6A-QTLs with disease genetics, including heart/muscle m6A-QTLs underlying coronary artery disease. Then, they had some remarkable results: coronary-artery-disease-associated rs888298 is a muscle/heart m6A-QTL targeting the cardiac myocyte mitochondrial oxidation regulator WIPI1 involved in signaling and autophagy. Heart-pulse-rate-associated rs6791834 is a muscle/heart m6A-QTL for myocyte microtubule differentiation regulator MAP4 involved in heart development. High-blood-pressure-associated rs56104944 is a muscle/heart m6A-QTL for heat shock protein HSPA4 involved in cardiac hypertrophy and fibrosis. Muscle/heart m6A-QTLs were enriched for 8 traits related to CVDs, reinforcing the roles of m6A regulators (FTO, METTL3, and ALKBH5) in CVDs. Li et al. utilized the quantitative trait loci of m6A, DNA methylation, and H3K27ac as genetic instruments to delineate locus-specific, directional maps of the crosstalk between m6A and the two fundamental epigenomic traits [104]. In muscle tissue, Li et al. uncovered 58 genetic loci that show strong multi-colocalization effects with the m6A-to-DNAme regulatory pairs, explaining GWAS traits such as high blood pressure and pulse rate. They also noticed that the variant rs12866090, associated with atrial fibrillation, likely mediates the crosstalk between m6A and DNA methylation in CUL4A.
Current evidence-based research unequivocally demonstrates that the integration of m6A with artificial intelligence and machine learning is highly significant for the prediction and diagnosis of CVDs. We anticipate that soon there will be more advanced methods with reduced error and false-positive rates, which will enhance our comprehension of the regulatory function of m6A in CVDs and offer new targets for forthcoming clinical interventions.

4. Discussion

This review provides a comprehensive summary of the pathophysiological effects of m6A methylation in cardiovascular risk factors, ischemia–hypoxia injury, atherosclerosis, myocardial infarction (MI), heart failure, and other cardiovascular diseases (Table 2 and Table 3). We also explored potential mechanisms of m6A methylation, including glucose metabolism, adipogenesis, VSMC differentiation and angiogenesis, autophagy, macrophage response, and inflammation, in the regulation of cardiovascular disease development.
However, m6A research is still in its early stages, and many unanswered questions remain. Currently, research on m6A methylation detection in cardiovascular diseases primarily focuses on the expression of METTL3 and FTO. Limited attention has been given to the effects of m6A on the cardiovascular system through LncRNA, circRNA, and miRNA. This could be attributed to the relatively lower expression of methylated regulatory enzymes in the cardiovascular system compared to other systems (Figure 3). Future research should aim to investigate how other methylases and demethylases regulate the expression of downstream proteins under various pathological factors. Additionally, the role and mechanism of m6A-binding proteins in mediating the activity of m6A methylation in cardiovascular diseases should be explored.
Furthermore, CVDs are complex and multifactorial, involving multiple mechanisms. It is important to explore the synergistic effects of multiple methylated mRNA pathways on pathological progression. Dorn and Kmietczyk’s conflicting conclusions regarding the inhibitory effect of METTL3 on cardiomyocyte hypertrophy suggest that changes in methylation patterns at different stages of disease occurrence and development may lead to distinct pathological processes. This highlights the need for further research on m6A-mediated methods for disease control, prevention, and treatment. Additionally, the effective clinical translation of m6A methylation findings is a topic that warrants comprehensive investigation [92,93].

5. Conclusions

M6A methylation participates extensively in diverse cardiovascular diseases. During the occurrence and progression of these diseases, proteins related to m6A methylation are also implicated in heart failure, myocardial hypertrophy, atherosclerosis, ischemic cardiomyopathy, and other conditions. It exerts a crucial regulatory function in the pathogenesis of these diseases. Moreover, the specificity of m6A varies. M6A methyltransferase can be applied in the early diagnosis of cardiovascular diseases. The dynamic changes in the expression and activity levels of METTL3 and METTL14, as well as the m6A demethylase FTO, can serve as potential biomarkers for cardiovascular diseases.

Author Contributions

Conceptualization, X.G. and J.Z.; formal analysis, A.C.; resources, J.Z.; writing—original draft preparation, H.Z. and W.L.; writing—review and editing, H.T., X.G., C.Z. and J.Z.; supervision, J.Z.; funding acquisition, X.G. and J.Z. All authors have read and agreed to the published version of the manuscript.

Funding

The study was supported by the National Natural Science Foundation of China (NSFC 81970307, NSFC 82100357, NSFC 82200385, and NSFC 82270344) and was jointly supported by the Jiangsu Provincial Medical Innovation Center (CXZX202215), the Jiangsu Natural Science Foundation (BK20221173), and the Nanjing Health Committee (JQX22007, and ZKX23030).

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Kottakis, F.; Nicolay, B.N.; Roumane, A.; Karnik, R.; Gu, H.; Nagle, J.M.; Boukhali, M.; Hayward, M.C.; Li, Y.Y.; Chen, T.; et al. LKB1 loss links serine metabolism to DNA methylation and tumorigenesis. Nature 2016, 539, 390–395. [Google Scholar] [CrossRef] [PubMed]
  2. Zaccara, S.; Ries, R.J.; Jaffrey, S.R. Reading, writing and erasing mRNA methylation. Nat. Rev. Mol. Cell Biol. 2019, 20, 608–624. [Google Scholar] [CrossRef] [PubMed]
  3. Wang, C.; Hou, X.; Guan, Q.; Zhou, H.; Zhou, L.; Liu, L.; Liu, J.; Li, F.; Li, W.; Liu, H. RNA modification in cardiovascular disease: Implications for therapeutic interventions. Signal Transduct. Target. Ther. 2023, 8, 412. [Google Scholar] [CrossRef] [PubMed]
  4. Motorin, Y.; Helm, M. RNA nucleotide methylation. Wiley Interdiscip. Rev. RNA 2011, 2, 611–631. [Google Scholar] [CrossRef]
  5. Desrosiers, R.; Friderici, K.; Rottman, F. Identification of methylated nucleosides in messenger RNA from Novikoff hepatoma cells. Proc. Natl. Acad. Sci. USA 1974, 71, 3971–3975. [Google Scholar] [CrossRef]
  6. Davalos, V.; Blanco, S.; Esteller, M. SnapShot: Messenger RNA Modifications. Cell 2018, 174, 498. [Google Scholar] [CrossRef]
  7. Dominissini, D.; Moshitch-Moshkovitz, S.; Schwartz, S.; Salmon-Divon, M.; Ungar, L.; Osenberg, S.; Cesarkas, K.; Jacob-Hirsch, J.; Amariglio, N.; Kupiec, M.; et al. Topology of the human and mouse m6A RNA methylomes revealed by m6A-seq. Nature 2012, 485, 201–206. [Google Scholar] [CrossRef]
  8. Han, J.; Wang, J.Z.; Yang, X.; Yu, H.; Zhou, R.; Lu, H.C.; Yuan, W.B.; Lu, J.C.; Zhou, Z.J.; Lu, Q.; et al. METTL3 promote tumor proliferation of bladder cancer by accelerating pri-miR221/222 maturation in m6A-dependent manner. Mol. Cancer 2019, 18, 110. [Google Scholar] [CrossRef]
  9. Zhou, C.; Molinie, B.; Daneshvar, K.; Pondick, J.V.; Wang, J.; Van Wittenberghe, N.; Xing, Y.; Giallourakis, C.C.; Mullen, A.C. Genome-Wide Maps of m6A circRNAs Identify Widespread and Cell-Type-Specific Methylation Patterns that Are Distinct from mRNAs. Cell Rep. 2017, 20, 2262–2276. [Google Scholar] [CrossRef]
  10. Yang, D.; Qiao, J.; Wang, G.; Lan, Y.; Li, G.; Guo, X.; Xi, J.; Ye, D.; Zhu, S.; Chen, W.; et al. N6-Methyladenosine modification of lincRNA 1281 is critically required for mESC differentiation potential. Nucleic Acids Res. 2018, 46, 3906–3920. [Google Scholar] [CrossRef]
  11. Noale, M.; Limongi, F.; Maggi, S. Epidemiology of Cardiovascular Diseases in the Elderly. Adv. Exp. Med. Biol. 2020, 1216, 29–38. [Google Scholar] [CrossRef] [PubMed]
  12. Meyer, K.D.; Saletore, Y.; Zumbo, P.; Elemento, O.; Mason, C.E.; Jaffrey, S.R. Comprehensive analysis of mRNA methylation reveals enrichment in 3′ UTRs and near stop codons. Cell 2012, 149, 1635–1646. [Google Scholar] [CrossRef] [PubMed]
  13. Yue, Y.; Liu, J.; He, C. RNA N6-methyladenosine methylation in post-transcriptional gene expression regulation. Genes. Dev. 2015, 29, 1343–1355. [Google Scholar] [CrossRef] [PubMed]
  14. Meyer, K.D.; Jaffrey, S.R. Rethinking m6A Readers, Writers, and Erasers. Annu. Rev. Cell Dev. Biol. 2017, 33, 319–342. [Google Scholar] [CrossRef] [PubMed]
  15. Yun, S.; He, X.; Zhang, W.; Chu, D.; Feng, C. Alleviation Effect of Grape Seed Proanthocyanidins on Neuronal Apoptosis in Rats with Iron Overload. Biol. Trace Elem. Res. 2020, 194, 210–220. [Google Scholar] [CrossRef]
  16. Liu, J.; Yue, Y.; Han, D.; Wang, X.; Fu, Y.; Zhang, L.; Jia, G.; Yu, M.; Lu, Z.; Deng, X.; et al. A METTL3-METTL14 complex mediates mammalian nuclear RNA N6-adenosine methylation. Nat. Chem. Biol. 2014, 10, 93–95. [Google Scholar] [CrossRef]
  17. Ping, X.L.; Sun, B.F.; Wang, L.; Xiao, W.; Yang, X.; Wang, W.J.; Adhikari, S.; Shi, Y.; Lv, Y.; Chen, Y.S.; et al. Mammalian WTAP is a regulatory subunit of the RNA N6-methyladenosine methyltransferase. Cell Res. 2014, 24, 177–189. [Google Scholar] [CrossRef]
  18. Schwartz, S.; Mumbach, M.R.; Jovanovic, M.; Wang, T.; Maciag, K.; Bushkin, G.G.; Mertins, P.; Ter-Ovanesyan, D.; Habib, N.; Cacchiarelli, D.; et al. Perturbation of m6A writers reveals two distinct classes of mRNA methylation at internal and 5′ sites. Cell Rep. 2014, 8, 284–296. [Google Scholar] [CrossRef]
  19. Patil, D.P.; Chen, C.K.; Pickering, B.F.; Chow, A.; Jackson, C.; Guttman, M.; Jaffrey, S.R. m6A RNA methylation promotes XIST-mediated transcriptional repression. Nature 2016, 537, 369–373. [Google Scholar] [CrossRef]
  20. Wen, J.; Lv, R.; Ma, H.; Shen, H.; He, C.; Wang, J.; Jiao, F.; Liu, H.; Yang, P.; Tan, L.; et al. Zc3h13 Regulates Nuclear RNA m6A Methylation and Mouse Embryonic Stem Cell Self-Renewal. Mol. Cell 2018, 69, 1028–1038.e6. [Google Scholar] [CrossRef]
  21. Yue, Y.; Liu, J.; Cui, X.; Cao, J.; Luo, G.; Zhang, Z.; Cheng, T.; Gao, M.; Shu, X.; Ma, H.; et al. VIRMA mediates preferential m6A mRNA methylation in 3′UTR and near stop codon and associates with alternative polyadenylation. Cell Discov. 2018, 4, 10. [Google Scholar] [CrossRef] [PubMed]
  22. Jia, G.; Fu, Y.; Zhao, X.; Dai, Q.; Zheng, G.; Yang, Y.; Yi, C.; Lindahl, T.; Pan, T.; Yang, Y.G.; et al. N6-methyladenosine in nuclear RNA is a major substrate of the obesity-associated FTO. Nat. Chem. Biol. 2011, 7, 885–887. [Google Scholar] [CrossRef] [PubMed]
  23. Zheng, G.; Dahl, J.A.; Niu, Y.; Fedorcsak, P.; Huang, C.M.; Li, C.J.; Vagbo, C.B.; Shi, Y.; Wang, W.L.; Song, S.H.; et al. ALKBH5 is a mammalian RNA demethylase that impacts RNA metabolism and mouse fertility. Mol. Cell 2013, 49, 18–29. [Google Scholar] [CrossRef] [PubMed]
  24. Wang, X.; Zhao, B.S.; Roundtree, I.A.; Lu, Z.; Han, D.; Ma, H.; Weng, X.; Chen, K.; Shi, H.; He, C. N6-methyladenosine Modulates Messenger RNA Translation Efficiency. Cell 2015, 161, 1388–1399. [Google Scholar] [CrossRef]
  25. Shi, H.; Wang, X.; Lu, Z.; Zhao, B.S.; Ma, H.; Hsu, P.J.; Liu, C.; He, C. YTHDF3 facilitates translation and decay of N6-methyladenosine-modified RNA. Cell Res. 2017, 27, 315–328. [Google Scholar] [CrossRef]
  26. Wang, X.; Lu, Z.; Gomez, A.; Hon, G.C.; Yue, Y.; Han, D.; Fu, Y.; Parisien, M.; Dai, Q.; Jia, G.; et al. N6-methyladenosine-dependent regulation of messenger RNA stability. Nature 2014, 505, 117–120. [Google Scholar] [CrossRef]
  27. Roundtree, I.A.; Luo, G.Z.; Zhang, Z.; Wang, X.; Zhou, T.; Cui, Y.; Sha, J.; Huang, X.; Guerrero, L.; Xie, P.; et al. YTHDC1 mediates nuclear export of N6-methyladenosine methylated mRNAs. eLife 2017, 6, e31311. [Google Scholar] [CrossRef]
  28. Hsu, P.J.; Zhu, Y.; Ma, H.; Guo, Y.; Shi, X.; Liu, Y.; Qi, M.; Lu, Z.; Shi, H.; Wang, J.; et al. Ythdc2 is an N6-methyladenosine binding protein that regulates mammalian spermatogenesis. Cell Res. 2017, 27, 1115–1127. [Google Scholar] [CrossRef]
  29. Degrauwe, N.; Suva, M.L.; Janiszewska, M.; Riggi, N.; Stamenkovic, I. IMPs: An RNA-binding protein family that provides a link between stem cell maintenance in normal development and cancer. Genes. Dev. 2016, 30, 2459–2474. [Google Scholar] [CrossRef]
  30. Wu, B.; Su, S.; Patil, D.P.; Liu, H.; Gan, J.; Jaffrey, S.R.; Ma, J. Molecular basis for the specific and multivariant recognitions of RNA substrates by human hnRNP A2/B1. Nat. Commun. 2018, 9, 420. [Google Scholar] [CrossRef]
  31. Tang, Y.; Chen, K.; Song, B.; Ma, J.; Wu, X.; Xu, Q.; Wei, Z.; Su, J.; Liu, G.; Rong, R.; et al. m6A-Atlas: A comprehensive knowledgebase for unraveling the N6-methyladenosine (m6A) epitranscriptome. Nucleic Acids Res. 2021, 49, D134–D143. [Google Scholar] [CrossRef] [PubMed]
  32. Li, W.; Gao, Y.; Jin, X.; Wang, H.; Lan, T.; Wei, M.; Yan, W.; Wang, G.; Li, Z.; Zhao, Z.; et al. Comprehensive analysis of N6-methylandenosine regulators and m6A-related RNAs as prognosis factors in colorectal cancer. Mol. Ther. Nucleic Acids 2022, 27, 598–610. [Google Scholar] [CrossRef] [PubMed]
  33. Wang, Y.; Li, Y.; Toth, J.I.; Petroski, M.D.; Zhang, Z.; Zhao, J.C. N6-methyladenosine modification destabilizes developmental regulators in embryonic stem cells. Nat. Cell Biol. 2014, 16, 191–198. [Google Scholar] [CrossRef] [PubMed]
  34. Xiao, W.; Adhikari, S.; Dahal, U.; Chen, Y.S.; Hao, Y.J.; Sun, B.F.; Sun, H.Y.; Li, A.; Ping, X.L.; Lai, W.Y.; et al. Nuclear m6A Reader YTHDC1 Regulates mRNA Splicing. Mol. Cell 2016, 61, 507–519. [Google Scholar] [CrossRef]
  35. Yang, Y.; Shen, F.; Huang, W.; Qin, S.; Huang, J.T.; Sergi, C.; Yuan, B.F.; Liu, S.M. Glucose Is Involved in the Dynamic Regulation of m6A in Patients with Type 2 Diabetes. J. Clin. Endocrinol. Metab. 2019, 104, 665–673. [Google Scholar] [CrossRef]
  36. De Jesus, D.F.; Zhang, Z.; Kahraman, S.; Brown, N.K.; Chen, M.; Hu, J.; Gupta, M.K.; He, C.; Kulkarni, R.N. m6A mRNA Methylation Regulates Human beta-Cell Biology in Physiological States and in Type 2 Diabetes. Nat. Metab. 2019, 1, 765–774. [Google Scholar] [CrossRef]
  37. Kruger, N.; Biwer, L.A.; Good, M.E.; Ruddiman, C.A.; Wolpe, A.G.; DeLalio, L.J.; Murphy, S.; Macal, E.H., Jr.; Ragolia, L.; Serbulea, V.; et al. Loss of Endothelial FTO Antagonizes Obesity-Induced Metabolic and Vascular Dysfunction. Circ. Res. 2020, 126, 232–242. [Google Scholar] [CrossRef]
  38. Wu, R.; Guo, G.; Bi, Z.; Liu, Y.; Zhao, Y.; Chen, N.; Wang, F.; Wang, Y.; Wang, X. m6A methylation modulates adipogenesis through JAK2-STAT3-C/EBPbeta signaling. Biochim. Biophys. Acta Gene Regul. Mech. 2019, 1862, 796–806. [Google Scholar] [CrossRef]
  39. Zhao, X.; Yang, Y.; Sun, B.F.; Shi, Y.; Yang, X.; Xiao, W.; Hao, Y.J.; Ping, X.L.; Chen, Y.S.; Wang, W.J.; et al. FTO-dependent demethylation of N6-methyladenosine regulates mRNA splicing and is required for adipogenesis. Cell Res. 2014, 24, 1403–1419. [Google Scholar] [CrossRef]
  40. Wang, X.; Wu, R.; Liu, Y.; Zhao, Y.; Bi, Z.; Yao, Y.; Liu, Q.; Shi, H.; Wang, F.; Wang, Y. m6A mRNA methylation controls autophagy and adipogenesis by targeting Atg5 and Atg7. Autophagy 2020, 16, 1221–1235. [Google Scholar] [CrossRef]
  41. Zhang, B.; Jiang, H.; Wu, J.; Cai, Y.; Dong, Z.; Zhao, Y.; Hu, Q.; Hu, K.; Sun, A.; Ge, J. m6A demethylase FTO attenuates cardiac dysfunction by regulating glucose uptake and glycolysis in mice with pressure overload-induced heart failure. Signal Transduct. Target. Ther. 2021, 6, 377. [Google Scholar] [CrossRef] [PubMed]
  42. Birsoy, K.; Chen, Z.; Friedman, J. Transcriptional regulation of adipogenesis by KLF4. Cell Metab. 2008, 7, 339–347. [Google Scholar] [CrossRef] [PubMed]
  43. Song, T.; Yang, Y.; Wei, H.; Xie, X.; Lu, J.; Zeng, Q.; Peng, J.; Zhou, Y.; Jiang, S.; Peng, J. Zfp217 mediates m6A mRNA methylation to orchestrate transcriptional and post-transcriptional regulation to promote adipogenic differentiation. Nucleic Acids Res. 2019, 47, 6130–6144. [Google Scholar] [CrossRef] [PubMed]
  44. Zhou, T.; Han, D.; Liu, J.; Shi, J.; Zhu, P.; Wang, Y.; Dong, N. Factors influencing osteogenic differentiation of human aortic valve interstitial cells. J. Thorac. Cardiovasc. Surg. 2021, 161, e163–e185. [Google Scholar] [CrossRef]
  45. Chen, J.; Ning, Y.; Zhang, H.; Song, N.; Gu, Y.; Shi, Y.; Cai, J.; Ding, X.; Zhang, X. METTL14-dependent m6A regulates vascular calcification induced by indoxyl sulfate. Life Sci. 2019, 239, 117034. [Google Scholar] [CrossRef]
  46. Song, H.; Feng, X.; Zhang, H.; Luo, Y.; Huang, J.; Lin, M.; Jin, J.; Ding, X.; Wu, S.; Huang, H.; et al. METTL3 and ALKBH5 oppositely regulate m6A modification of TFEB mRNA, which dictates the fate of hypoxia/reoxygenation-treated cardiomyocytes. Autophagy 2019, 15, 1419–1437. [Google Scholar] [CrossRef]
  47. Kumari, R.; Dutta, R.; Ranjan, P.; Suleiman, Z.G.; Goswami, S.K.; Li, J.; Pal, H.C.; Verma, S.K. ALKBH5 Regulates SPHK1-Dependent Endothelial Cell Angiogenesis Following Ischemic Stress. Front. Cardiovasc. Med. 2021, 8, 817304. [Google Scholar] [CrossRef]
  48. Han, Z.; Wang, X.; Xu, Z.; Cao, Y.; Gong, R.; Yu, Y.; Yu, Y.; Guo, X.; Liu, S.; Yu, M.; et al. ALKBH5 regulates cardiomyocyte proliferation and heart regeneration by demethylating the mRNA of YTHDF1. Theranostics 2021, 11, 3000–3016. [Google Scholar] [CrossRef]
  49. Zhao, Y.; Hu, J.; Sun, X.; Yang, K.; Yang, L.; Kong, L.; Zhang, B.; Li, F.; Li, C.; Shi, B.; et al. Loss of m6A demethylase ALKBH5 promotes post-ischemic angiogenesis via post-transcriptional stabilization of WNT5A. Clin. Transl. Med. 2021, 11, e402. [Google Scholar] [CrossRef]
  50. Shen, W.; Li, H.; Su, H.; Chen, K.; Yan, J. FTO overexpression inhibits apoptosis of hypoxia/reoxygenation-treated myocardial cells by regulating m6A modification of Mhrt. Mol. Cell Biochem. 2021, 476, 2171–2179. [Google Scholar] [CrossRef]
  51. Yin, T.; Wang, N.; Jia, F.; Wu, Y.; Gao, L.; Zhang, J.; Hou, R. Exosome-based WTAP siRNA delivery ameliorates myocardial ischemia-reperfusion injury. Eur. J. Pharm. Biopharm. 2024, 197, 114218. [Google Scholar] [CrossRef] [PubMed]
  52. Chien, C.S.; Li, J.Y.; Chien, Y.; Wang, M.L.; Yarmishyn, A.A.; Tsai, P.H.; Juan, C.C.; Nguyen, P.; Cheng, H.M.; Huo, T.I.; et al. METTL3-dependent N6-methyladenosine RNA modification mediates the atherogenic inflammatory cascades in vascular endothelium. Proc. Natl. Acad. Sci. USA 2021, 118, e2025070118. [Google Scholar] [CrossRef] [PubMed]
  53. Jian, D.; Wang, Y.; Jian, L.; Tang, H.; Rao, L.; Chen, K.; Jia, Z.; Zhang, W.; Liu, Y.; Chen, X.; et al. METTL14 aggravates endothelial inflammation and atherosclerosis by increasing FOXO1 N6-methyladeosine modifications. Theranostics 2020, 10, 8939–8956. [Google Scholar] [CrossRef] [PubMed]
  54. Dong, G.; Yu, J.; Shan, G.; Su, L.; Yu, N.; Yang, S. N6-Methyladenosine Methyltransferase METTL3 Promotes Angiogenesis and Atherosclerosis by Upregulating the JAK2/STAT3 Pathway via m6A Reader IGF2BP1. Front. Cell Dev. Biol. 2021, 9, 731810. [Google Scholar] [CrossRef]
  55. Zhang, B.Y.; Han, L.; Tang, Y.F.; Zhang, G.X.; Fan, X.L.; Zhang, J.J.; Xue, Q.; Xu, Z.Y. METTL14 regulates M6A methylation-modified primary miR-19a to promote cardiovascular endothelial cell proliferation and invasion. Eur. Rev. Med. Pharmacol. Sci. 2020, 24, 7015–7023. [Google Scholar] [CrossRef]
  56. Huo, Y.B.; Gao, X.; Peng, Q.; Nie, Q.; Bi, W. Dihydroartemisinin alleviates AngII-induced vascular smooth muscle cell proliferation and inflammatory response by blocking the FTO/NR4A3 axis. Inflamm. Res. 2022, 71, 243–253. [Google Scholar] [CrossRef]
  57. Zhu, B.; Gong, Y.; Shen, L.; Li, J.; Han, J.; Song, B.; Hu, L.; Wang, Q.; Wang, Z. Total Panax notoginseng saponin inhibits vascular smooth muscle cell proliferation and migration and intimal hyperplasia by regulating WTAP/p16 signals via m6A modulation. Biomed. Pharmacother. 2020, 124, 109935. [Google Scholar] [CrossRef]
  58. Gong, R.; Wang, X.; Li, H.; Liu, S.; Jiang, Z.; Zhao, Y.; Yu, Y.; Han, Z.; Yu, Y.; Dong, C.; et al. Loss of m6A methyltransferase METTL3 promotes heart regeneration and repair after myocardial injury. Pharmacol. Res. 2021, 174, 105845. [Google Scholar] [CrossRef]
  59. Cheng, P.; Han, H.; Chen, F.; Cheng, L.; Ma, C.; Huang, H.; Chen, C.; Li, H.; Cai, H.; Huang, H.; et al. Amelioration of acute myocardial infarction injury through targeted ferritin nanocages loaded with an ALKBH5 inhibitor. Acta Biomater. 2022, 140, 481–491. [Google Scholar] [CrossRef]
  60. Yang, K.; Zhao, Y.; Hu, J.; Gao, R.; Shi, J.; Wei, X.; Chen, J.; Hu, K.; Sun, A.; Ge, J. ALKBH5 induces fibroblast-to-myofibroblast transformation during hypoxia to protect against cardiac rupture after myocardial infarction. J. Adv. Res. 2024, 61, 193–209. [Google Scholar] [CrossRef]
  61. Pang, P.; Qu, Z.; Yu, S.; Pang, X.; Li, X.; Gao, Y.; Liu, K.; Liu, Q.; Wang, X.; Bian, Y.; et al. Mettl14 Attenuates Cardiac Ischemia/Reperfusion Injury by Regulating Wnt1/beta-Catenin Signaling Pathway. Front. Cell Dev. Biol. 2021, 9, 762853. [Google Scholar] [CrossRef] [PubMed]
  62. Mathiyalagan, P.; Adamiak, M.; Mayourian, J.; Sassi, Y.; Liang, Y.; Agarwal, N.; Jha, D.; Zhang, S.; Kohlbrenner, E.; Chepurko, E.; et al. FTO-Dependent N6-Methyladenosine Regulates Cardiac Function During Remodeling and Repair. Circulation 2019, 139, 518–532. [Google Scholar] [CrossRef]
  63. Xu, H.; Wang, Z.; Chen, M.; Zhao, W.; Tao, T.; Ma, L.; Ni, Y.; Li, W. YTHDF2 alleviates cardiac hypertrophy via regulating Myh7 mRNA decoy. Cell Biosci. 2021, 11, 132. [Google Scholar] [CrossRef] [PubMed]
  64. Qian, B.; Wang, P.; Zhang, D.; Wu, L. m6A modification promotes miR-133a repression during cardiac development and hypertrophy via IGF2BP2. Cell Death Discov. 2021, 7, 157. [Google Scholar] [CrossRef] [PubMed]
  65. Zhuang, T.; Chen, M.H.; Wu, R.X.; Wang, J.; Hu, X.D.; Meng, T.; Wu, A.H.; Li, Y.; Yang, Y.F.; Lei, Y.; et al. ALKBH5-mediated m6A modification of IL-11 drives macrophage-to-myofibroblast transition and pathological cardiac fibrosis in mice. Nat. Commun. 2024, 15, 1995. [Google Scholar] [CrossRef]
  66. Qin, Y.; Qiao, Y.; Li, L.; Luo, E.; Wang, D.; Yao, Y.; Tang, C.; Yan, G. The m6A methyltransferase METTL3 promotes hypoxic pulmonary arterial hypertension. Life Sci. 2021, 274, 119366. [Google Scholar] [CrossRef]
  67. Hu, L.; Wang, J.; Huang, H.; Yu, Y.; Ding, J.; Yu, Y.; Li, K.; Wei, D.; Ye, Q.; Wang, F.; et al. YTHDF1 Regulates Pulmonary Hypertension through Translational Control of MAGED1. Am. J. Respir. Crit. Care Med. 2021, 203, 1158–1172. [Google Scholar] [CrossRef]
  68. Zhong, L.; He, X.; Song, H.; Sun, Y.; Chen, G.; Si, X.; Sun, J.; Chen, X.; Liao, W.; Liao, Y.; et al. METTL3 Induces AAA Development and Progression by Modulating N6-Methyladenosine-Dependent Primary miR34a Processing. Mol. Ther. Nucleic Acids 2020, 21, 394–411. [Google Scholar] [CrossRef]
  69. Wang, X.; Li, Y.; Li, J.; Li, S.; Wang, F. Mechanism of METTL3-Mediated m6A Modification in Cardiomyocyte Pyroptosis and Myocardial Ischemia-Reperfusion Injury. Cardiovasc. Drugs Ther. 2023, 37, 435–448. [Google Scholar] [CrossRef]
  70. Kroemer, G. Autophagy: A druggable process that is deregulated in aging and human disease. J. Clin. Investig. 2015, 125, 1–4. [Google Scholar] [CrossRef]
  71. Jin, S.; Zhang, X.; Miao, Y.; Liang, P.; Zhu, K.; She, Y.; Wu, Y.; Liu, D.A.; Huang, J.; Ren, J.; et al. m6A RNA modification controls autophagy through upregulating ULK1 protein abundance. Cell Res. 2018, 28, 955–957. [Google Scholar] [CrossRef] [PubMed]
  72. Frismantiene, A.; Philippova, M.; Erne, P.; Resink, T.J. Smooth muscle cell-driven vascular diseases and molecular mechanisms of VSMC plasticity. Cell Signal 2018, 52, 48–64. [Google Scholar] [CrossRef]
  73. Lin, J.; Zhu, Q.; Huang, J.; Cai, R.; Kuang, Y. Hypoxia Promotes Vascular Smooth Muscle Cell (VSMC) Differentiation of Adipose-Derived Stem Cell (ADSC) by Regulating Mettl3 and Paracrine Factors. Stem Cells Int. 2020, 2020, 2830565. [Google Scholar] [CrossRef] [PubMed]
  74. Yuan, J.; Liu, Y.; Zhou, L.; Xue, Y.; Lu, Z.; Gan, J. YTHDC2-Mediated circYTHDC2 N6-Methyladenosine Modification Promotes Vascular Smooth Muscle Cells Dysfunction Through Inhibiting Ten-Eleven Translocation 2. Front. Cardiovasc. Med. 2021, 8, 686293. [Google Scholar] [CrossRef] [PubMed]
  75. Parial, R.; Li, H.; Li, J.; Archacki, S.; Yang, Z.; Wang, I.Z.; Chen, Q.; Xu, C.; Wang, Q.K. Role of epigenetic m6A RNA methylation in vascular development: Mettl3 regulates vascular development through PHLPP2/mTOR-AKT signaling. FASEB J. 2021, 35, e21465. [Google Scholar] [CrossRef] [PubMed]
  76. Yao, M.D.; Jiang, Q.; Ma, Y.; Liu, C.; Zhu, C.Y.; Sun, Y.N.; Shan, K.; Ge, H.M.; Zhang, Q.Y.; Zhang, H.Y.; et al. Role of METTL3-Dependent N6-Methyladenosine mRNA Modification in the Promotion of Angiogenesis. Mol. Ther. 2020, 28, 2191–2202. [Google Scholar] [CrossRef]
  77. Geovanini, G.R.; Libby, P. Atherosclerosis and inflammation: Overview and updates. Clin. Sci. 2018, 132, 1243–1252. [Google Scholar] [CrossRef]
  78. Chiu, J.J.; Chien, S. Effects of disturbed flow on vascular endothelium: Pathophysiological basis and clinical perspectives. Physiol. Rev. 2011, 91, 327–387. [Google Scholar] [CrossRef]
  79. Miano, J.M.; Fisher, E.A.; Majesky, M.W. Fate and State of Vascular Smooth Muscle Cells in Atherosclerosis. Circulation 2021, 143, 2110–2116. [Google Scholar] [CrossRef]
  80. Libby, P. The changing landscape of atherosclerosis. Nature 2021, 592, 524–533. [Google Scholar] [CrossRef]
  81. Shapouri-Moghaddam, A.; Mohammadian, S.; Vazini, H.; Taghadosi, M.; Esmaeili, S.A.; Mardani, F.; Seifi, B.; Mohammadi, A.; Afshari, J.T.; Sahebkar, A. Macrophage plasticity, polarization, and function in health and disease. J. Cell Physiol. 2018, 233, 6425–6440. [Google Scholar] [CrossRef] [PubMed]
  82. Liu, Y.; Liu, Z.; Tang, H.; Shen, Y.; Gong, Z.; Xie, N.; Zhang, X.; Wang, W.; Kong, W.; Zhou, Y.; et al. The N6-methyladenosine (m6A)-forming enzyme METTL3 facilitates M1 macrophage polarization through the methylation of STAT1 mRNA. Am. J. Physiol. Cell Physiol. 2019, 317, C762–C775. [Google Scholar] [CrossRef] [PubMed]
  83. Wang, H.; Hu, X.; Huang, M.; Liu, J.; Gu, Y.; Ma, L.; Zhou, Q.; Cao, X. Mettl3-mediated mRNA m6A methylation promotes dendritic cell activation. Nat. Commun. 2019, 10, 1898. [Google Scholar] [CrossRef] [PubMed]
  84. Zhu, H.; Gan, X.; Jiang, X.; Diao, S.; Wu, H.; Hu, J. ALKBH5 inhibited autophagy of epithelial ovarian cancer through miR-7 and BCL-2. J. Exp. Clin. Cancer Res. 2019, 38, 163. [Google Scholar] [CrossRef]
  85. Writing Committee, M.; Yancy, C.W.; Jessup, M.; Bozkurt, B.; Butler, J.; Casey, D.E., Jr.; Drazner, M.H.; Fonarow, G.C.; Geraci, S.A.; Horwich, T.; et al. 2013 ACCF/AHA guideline for the management of heart failure: A report of the American College of Cardiology Foundation/American Heart Association Task Force on practice guidelines. Circulation 2013, 128, e240–e327. [Google Scholar] [CrossRef]
  86. Roger, V.L.; Weston, S.A.; Redfield, M.M.; Hellermann-Homan, J.P.; Killian, J.; Yawn, B.P.; Jacobsen, S.J. Trends in heart failure incidence and survival in a community-based population. JAMA 2004, 292, 344–350. [Google Scholar] [CrossRef]
  87. Zhang, B.; Xu, Y.; Cui, X.; Jiang, H.; Luo, W.; Weng, X.; Wang, Y.; Zhao, Y.; Sun, A.; Ge, J. Alteration of m6A RNA Methylation in Heart Failure With Preserved Ejection Fraction. Front. Cardiovasc. Med. 2021, 8, 647806. [Google Scholar] [CrossRef]
  88. Berulava, T.; Buchholz, E.; Elerdashvili, V.; Pena, T.; Islam, M.R.; Lbik, D.; Mohamed, B.A.; Renner, A.; von Lewinski, D.; Sacherer, M.; et al. Changes in m6A RNA methylation contribute to heart failure progression by modulating translation. Eur. J. Heart Fail. 2020, 22, 54–66. [Google Scholar] [CrossRef]
  89. Hinger, S.A.; Wei, J.; Dorn, L.E.; Whitson, B.A.; Janssen, P.M.L.; He, C.; Accornero, F. Remodeling of the m6A landscape in the heart reveals few conserved post-transcriptional events underlying cardiomyocyte hypertrophy. J. Mol. Cell Cardiol. 2021, 151, 46–55. [Google Scholar] [CrossRef]
  90. Li, T.; Zhuang, Y.; Yang, W.; Xie, Y.; Shang, W.; Su, S.; Dong, X.; Wu, J.; Jiang, W.; Zhou, Y.; et al. Silencing of METTL3 attenuates cardiac fibrosis induced by myocardial infarction via inhibiting the activation of cardiac fibroblasts. FASEB J. 2021, 35, e21162. [Google Scholar] [CrossRef]
  91. Fang, M.; Deng, J.; Zhou, Q.; Hu, Z.; Yang, L. Maslinic acid protects against pressure-overload-induced cardiac hypertrophy by blocking METTL3-mediated m6A methylation. Aging 2022, 14, 2548–2557. [Google Scholar] [CrossRef] [PubMed]
  92. Dorn, L.E.; Lasman, L.; Chen, J.; Xu, X.; Hund, T.J.; Medvedovic, M.; Hanna, J.H.; van Berlo, J.H.; Accornero, F. The N6-Methyladenosine mRNA Methylase METTL3 Controls Cardiac Homeostasis and Hypertrophy. Circulation 2019, 139, 533–545. [Google Scholar] [CrossRef] [PubMed]
  93. Kmietczyk, V.; Riechert, E.; Kalinski, L.; Boileau, E.; Malovrh, E.; Malone, B.; Gorska, A.; Hofmann, C.; Varma, E.; Jurgensen, L.; et al. m6A-mRNA methylation regulates cardiac gene expression and cellular growth. Life Sci. Alliance 2019, 2, e201800233. [Google Scholar] [CrossRef] [PubMed]
  94. Xu, C.; Hu, Y.; Hou, L.; Ju, J.; Li, X.; Du, N.; Guan, X.; Liu, Z.; Zhang, T.; Qin, W.; et al. beta-Blocker carvedilol protects cardiomyocytes against oxidative stress-induced apoptosis by up-regulating miR-133 expression. J. Mol. Cell Cardiol. 2014, 75, 111–121. [Google Scholar] [CrossRef]
  95. Liu, N.; Bezprozvannaya, S.; Williams, A.H.; Qi, X.; Richardson, J.A.; Bassel-Duby, R.; Olson, E.N. microRNA-133a regulates cardiomyocyte proliferation and suppresses smooth muscle gene expression in the heart. Genes. Dev. 2008, 22, 3242–3254. [Google Scholar] [CrossRef]
  96. Marcadenti, A.; Fuchs, F.D.; Matte, U.; Sperb, F.; Moreira, L.B.; Fuchs, S.C. Effects of FTO RS9939906 and MC4R RS17782313 on obesity, type 2 diabetes mellitus and blood pressure in patients with hypertension. Cardiovasc. Diabetol. 2013, 12, 103. [Google Scholar] [CrossRef]
  97. Mo, X.B.; Lei, S.F.; Zhang, Y.H.; Zhang, H. Examination of the associations between m6A-associated single-nucleotide polymorphisms and blood pressure. Hypertens. Res. 2019, 42, 1582–1589. [Google Scholar] [CrossRef]
  98. Su, H.; Wang, G.; Wu, L.; Ma, X.; Ying, K.; Zhang, R. Transcriptome-wide map of m6A circRNAs identified in a rat model of hypoxia mediated pulmonary hypertension. BMC Genom. 2020, 21, 39. [Google Scholar] [CrossRef]
  99. Ma, D.; Liu, X.; Zhang, J.J.; Zhao, J.J.; Xiong, Y.J.; Chang, Q.; Wang, H.Y.; Su, P.; Meng, J.; Zhao, Y.B. Vascular Smooth Muscle FTO Promotes Aortic Dissecting Aneurysms via m6A Modification of Klf5. Front. Cardiovasc. Med. 2020, 7, 592550. [Google Scholar] [CrossRef]
  100. Li, T.; Wu, Y.; Yang, J.; Jing, J.; Ma, C.; Sun, L. N6-methyladenosine-associated genetic variants in NECTIN2 and HPCAL1 are risk factors for abdominal aortic aneurysm. iScience 2024, 27, 109419. [Google Scholar] [CrossRef]
  101. Huang, Y.; Yan, J.; Li, Q.; Li, J.; Gong, S.; Zhou, H.; Gan, J.; Jiang, H.; Jia, G.F.; Luo, C.; et al. Meclofenamic acid selectively inhibits FTO demethylation of m6A over ALKBH5. Nucleic Acids Res. 2015, 43, 373–384. [Google Scholar] [CrossRef] [PubMed]
  102. Ambale-Venkatesh, B.; Yang, X.; Wu, C.O.; Liu, K.; Hundley, W.G.; McClelland, R.; Gomes, A.S.; Folsom, A.R.; Shea, S.; Guallar, E.; et al. Cardiovascular Event Prediction by Machine Learning: The Multi-Ethnic Study of Atherosclerosis. Circ. Res. 2017, 121, 1092–1101. [Google Scholar] [CrossRef] [PubMed]
  103. Xiong, X.; Hou, L.; Park, Y.P.; Molinie, B.; Consortium, G.T.; Gregory, R.I.; Kellis, M. Genetic drivers of m6A methylation in human brain, lung, heart and muscle. Nat. Genet. 2021, 53, 1156–1165. [Google Scholar] [CrossRef] [PubMed]
  104. Li, C.; Chen, K.; Fang, Q.; Shi, S.; Nan, J.; He, J.; Yin, Y.; Li, X.; Li, J.; Hou, L.; et al. Crosstalk between epitranscriptomic and epigenomic modifications and its implication in human diseases. Cell Genom. 2024, 4, 100605. [Google Scholar] [CrossRef]
Medicina 61 00222 g001
Figure 1. The regulation of m6A modification. m6A modification is established by m6A methyltransferases (“writers”) and removed by m6A demethylases (“erasers”). m6A-binding proteins (“readers”) recognize and bind to m6A-modified RNA, playing essential roles in RNA metabolism. METTL3, in collaboration with METTL14/16, KIAA1429, WTAP, RBM15/15B, and ZC3H13, forms the core methylation complex. This modification is reversible, with demethylases such as ALKBH5 and FTO serving as m6A erasers. The modified transcripts are recognized by readers, including YTHDF1/2/3, YTHDC1/2, IGF2BP1/2/3, and hnRNPG/C/A2B, which subsequently influence various aspects of RNA function, such as translation promotion, stability, localization, splicing, and nuclear export.
Figure 1. The regulation of m6A modification. m6A modification is established by m6A methyltransferases (“writers”) and removed by m6A demethylases (“erasers”). m6A-binding proteins (“readers”) recognize and bind to m6A-modified RNA, playing essential roles in RNA metabolism. METTL3, in collaboration with METTL14/16, KIAA1429, WTAP, RBM15/15B, and ZC3H13, forms the core methylation complex. This modification is reversible, with demethylases such as ALKBH5 and FTO serving as m6A erasers. The modified transcripts are recognized by readers, including YTHDF1/2/3, YTHDC1/2, IGF2BP1/2/3, and hnRNPG/C/A2B, which subsequently influence various aspects of RNA function, such as translation promotion, stability, localization, splicing, and nuclear export.
Medicina 61 00222 g001
Medicina 61 00222 g002
Figure 2. The role of m6A modulators in cardiovascular diseases and biological processes. METTL3-mediated m6A modification contributes to atherosclerosis by affecting inflammatory pathways, including NLRP1, KLF4, and JAK2. METTL14 up-regulation induces inflammation and plaque formation by enhancing FOXO1 translation. Reduced FTO expression promotes smooth muscle cell proliferation and inflammation by stabilizing NR4A3 mRNA, accelerating atherosclerosis. In stent restenosis, WTAP promotes smooth muscle cell proliferation and migration by increasing P16 mRNA via m6A modification. In myocardial ischemic disease, METTL3 up-regulation decreases TFEB expression, impairing autophagic flux and enhancing cell apoptosis. During myocardial infarction repair, ALKBH5 up-regulation inhibits autophagy by destabilizing mRNAs like SPHK1, ERB4, and YAP, promoting infarct repair. METTL3 also mediates miR-34a maturation, which down-regulates SIRT1 and promotes inflammatory infiltration in abdominal aortic aneurysm. Decreased FTO expression is linked to reduced Serca2a levels, leading to impaired cardiac contractility and heart failure. In heart failure, m6A-modified MYH7 mRNA and miR-133a play protective roles in ventricular remodeling. Additionally, ALKBH5 increases IL-11 mRNA, inhibiting macrophage-to-myofibroblast transition. In hypoxic pulmonary hypertension, up-regulated MAGED1 and down-regulated PTEN, both m6A-modified, contribute to smooth muscle cell proliferation, inflammation, and pulmonary vascular remodeling.
Figure 2. The role of m6A modulators in cardiovascular diseases and biological processes. METTL3-mediated m6A modification contributes to atherosclerosis by affecting inflammatory pathways, including NLRP1, KLF4, and JAK2. METTL14 up-regulation induces inflammation and plaque formation by enhancing FOXO1 translation. Reduced FTO expression promotes smooth muscle cell proliferation and inflammation by stabilizing NR4A3 mRNA, accelerating atherosclerosis. In stent restenosis, WTAP promotes smooth muscle cell proliferation and migration by increasing P16 mRNA via m6A modification. In myocardial ischemic disease, METTL3 up-regulation decreases TFEB expression, impairing autophagic flux and enhancing cell apoptosis. During myocardial infarction repair, ALKBH5 up-regulation inhibits autophagy by destabilizing mRNAs like SPHK1, ERB4, and YAP, promoting infarct repair. METTL3 also mediates miR-34a maturation, which down-regulates SIRT1 and promotes inflammatory infiltration in abdominal aortic aneurysm. Decreased FTO expression is linked to reduced Serca2a levels, leading to impaired cardiac contractility and heart failure. In heart failure, m6A-modified MYH7 mRNA and miR-133a play protective roles in ventricular remodeling. Additionally, ALKBH5 increases IL-11 mRNA, inhibiting macrophage-to-myofibroblast transition. In hypoxic pulmonary hypertension, up-regulated MAGED1 and down-regulated PTEN, both m6A-modified, contribute to smooth muscle cell proliferation, inflammation, and pulmonary vascular remodeling.
Medicina 61 00222 g002
Medicina 61 00222 g003
Figure 3. Relative amounts of methylated regulatory enzymes in the cardiovascular system and other systems. Heatmap shows differentially expressed methylated regulatory enzymes in different tissues. Data derived from PRJEB4337 of HPA RNA-seq normal tissues, in which RNA-seq was performed on tissue samples from 95 human individuals representing 27 different tissues in order to determine the tissue specificity of all protein-coding genes.
Figure 3. Relative amounts of methylated regulatory enzymes in the cardiovascular system and other systems. Heatmap shows differentially expressed methylated regulatory enzymes in different tissues. Data derived from PRJEB4337 of HPA RNA-seq normal tissues, in which RNA-seq was performed on tissue samples from 95 human individuals representing 27 different tissues in order to determine the tissue specificity of all protein-coding genes.
Medicina 61 00222 g003
Table 1. Writers, erasers, and readers in m6A.
Table 1. Writers, erasers, and readers in m6A.
TypeRegulatorFunction of RNA ModificationReference
writersMETTL3main catalytic subunit of m6A[16]
METTL14/16activate METTL3 through allosteric and RNA substrate recognition[16]
WTAPthe third subunit of the METTL3-METTL14 complex[17,18]
ZC3H3assist the localization of the methyltransferase complex in nuclear speckles and U-rich regions adjacent to the m6A sites in mRNAs[19]
RBM15/15B[20]
KIAA1429[21]
erasersFTOdemethylation of m6a[2,22,23]
ALKBH5
readersYTHDF1promote mRNA translation[24,25]
YTHDF2accelerate the decay of m6A-modified transcripts
YTHDF3promote mRNA translation or enhance RNA decay
YTHDC1promote mRNA translation and splicing and nuclear export[26,27]
YTHDC2enhance translation[28]
IGF2BP1/2/3regulate RNA localization, translation, and stability[29]
hnRNPG/C/A2Bpromote RNA stability and mediate RNA splicing and microRNA process[30]
Abbreviations: ALKBH5: ALKB homolog 5, RNA demethylase; FTO: FTO alpha-ketoglutarate dependent dioxygenase; KIAA1429: vir like m6A methyltransferase associated; METTL3: methyltransferase3; METTL14: methyltransferase14; METTL16: methyltransferase16; RBM15/15B: RNA binding motif protein 15B; WTAP: WT1associated protein; YTHDF1: YTH N6-methyladenosine RNA binding protein F1; YTHDF2: YTH N6-methyladenosine RNA binding protein F2; YTHDF3: YTH N6-methyladenosine RNA binding protein F3; YTHDC1: YTH N6-methyladenosine RNA binding protein C1; YTHDC2: YTH N6-methyladenosine RNA binding protein C2; IGF2BP1/2/3: insulin like growth factor 2 mRNA binding protein 1/2/3; hnRNPG/C/A2B: RNA binding motif protein X-linked G/C/A2B; ZC3H3: zinc finger CCCH-type containing3.
Table 2. Molecular mechanisms of m6a in risk factors in CVDs.
Table 2. Molecular mechanisms of m6a in risk factors in CVDs.
Risk FactorsRegulatorsCellRegulationSignalingFunctionReference
glucose metabolismFTO↑hepatocellular cellup-regulate mRNAFOXO1/FASN/
G6PC/DGAT2		improve the production of serum glucose and lipids[35]
diabetesMETTL14↓pancreatic β-cellpromote mRNA translationAKT/PDX1induce cell-cycle arrest and impair insulin secretion[36]
obesityFTO↑endothelial celldown-regulate mRNAAKT/
prostaglandinD2		aggravate vascular dysfunction[37]
FTO↑preadipocyteup-regulate mRNAJAK2-STAT3-C/EBPβpromote adipogenesis[38]
FTO↑preadipocytecontrol exonic splicingRUNX1T1modulate differentiation to promote adipogenesis[39]
FTO↑preadipocytesimprove mRNA stabilityAtg5/Atg7promote autophagy and adipogenesis[40]
Abbreviations: FTO: FTO alpha-ketoglutarate dependent dioxygenase; METTL14:methyltransferase14; FOXO1: forkhead box O1; FASN: fatty acid synthase; G6PC: glucose-6-phosphatase catalytic subunit; DGAT2: diacylglycerol O-acyltransferase 2; AKT: Akt kinase; PDX1: pancreatic and duodenal homeobox 1; JAK2: Janus kinase 2; STAT3: signal transducer and activator of transcription 3; C/EBP: CCAAT/enhancer binding protein; RUNX1T1: RUNX1 partner transcriptional co-repressor 1; ATG5: autophagy related 5; ATG7: autophagy related 7.
Table 3. The roles of m6A methylation in CVDs.
Table 3. The roles of m6A methylation in CVDs.
DiseasesRegulatorsCellRegulationSignalingFunctionReference
calcificationMETTL3↑valve interstitial cellup-regulate mRNATWIST1promote osteogenic differentiation process[44]
calcificationMETTL14↑smooth muscle celldown-regulate mRNAKlothopromote calcification[45]
Hypoxia-reoxygenationMETTL3↑cardiomyocytedown-regulate mRNATFEBinhibit autophagy and enhance apoptosis [46]
ischemic injuryALKBH5↑endothelial cellup-regulate mRNASPHK1/eNOS-AKTmaintain angiogenesis [47]
heart regenerationALKBH5↑cardiomyocyteimprove mRNA stabilityYAPpromote proliferation[48]
post-ischemicALKBH5↑endothelial celldecrease mRNA stabilityWNT5Aexacerbate dysfunction of CMECs[49]
Hypoxia-reoxygenationFTO↑cardiomyocyteup-regulate mRNAMhrtinhibit apoptosis [50]
Hypoxia-reoxygenationWTAP↑cardiomyocyteup-regulate mRNATXNIPenhance apoptosis [51]
atherosclerosisMETTL3↑endothelial cellup/down-regulate mRNANLRP1/KLF4promote inflammatory cascades[52]
atherosclerosisMETTL14↑endothelial cellpromote translationFOXO1/VCAM-1/ICAM-1induce inflammatory response and promote atherosclerotic plaque formation[53]
atherosclerosisMETTL3↑endothelial cellup-regulate mRNAJAK2/STAT3promote atherosclerosis progression[54]
atherosclerosisMETTL14↑endothelial cellup-regulate miRNA pri-miR-19a
/DGCR8		promote proliferation and invasion of ASVEC[55]
atherosclerosisFTO↓smooth muscle cellup-regulate mRNANR4A3promote proliferation and inflammation [56]
intimal hyperplasiaWTAP↓smooth muscle cellup-regulate mRNAp16promote proliferation and migration of VSMC[57]
heart regenerationMETTL3↓cardiomyocyteup-regulate miRNA miR-143
/Yap/Ctnnd1		inhibit heart regeneration [58]
AMLALKBH5↑cardiomyocytenot mentionTCA cycleaffect cell metabolism and survival[59]
AMLALKBH5↑fibroblastimprove mRNA stabilityErbB4regulate post-MI healing [60]
Ischemia-reperfusion injuryMETTL14↑cardiomyocytepromote translation efficiencyWnt1attenuate ischemia–reperfusion Injury[61]
heart failureFTO↓cardiomyocyteimprove mRNA stabilitySerca2aimprove Ca2+ amplitude[62]
heart failureYTHDF2↑cardiomyocytepromote mRNA
degradation		Myh7alleviate cardiac hypertrophy[63]
heart failureIGF2BP2↑ cardiomyocytepromote miRNA accumulation on its target sitemiR-133a repress cardiac hypertrophy and apoptosis [64]
heart failureFTO↓cardiomyocytesimprove mRNA stabilityPgam2regulate glucose uptake[41]
heart failureALKBH5↑ macrophage improve mRNA stabilityIL-11inhibit macrophage-to-myofibroblast transition [65]
pulmonary hypertensionYTHDF2↑smooth muscle cellpromote mRNA
degradation		PTEN/PI3K/Aktenhance proliferation[66]
pulmonary hypertensionYTHDF1↑smooth muscle cellpromote translation efficiencyMAGED1promote proliferation[67]
aneurysmMETTL3↑smooth muscle cellup-regulate miRNA DGCR8/
miR-34a		induce development and progression[68]
Abbreviations: ALKBH5: ALKB homolog 5, RNA demethylase; FTO: FTO alpha-ketoglutarate dependent dioxygenase; FOXO1: forkhead box O1; ICAM-1: intercellular adhesion molecule 1; JAK2: Janus kinase 2; METTL3: methyltransferase3; METTL14:methyltransferase14; Myh7: myosin heavy chain 7; MAGED1: MAGE family member D1; NLRP1: NLR family pyrin domain containing 1; KLF4: KLF transcription factor 4; NR4A3: nuclear receptor subfamily 4 group A member 3; SPHK1: sphingosine kinase 1; STAT3: signal transducer and activator of transcription 3; Serca2a: Sarco/endoplasmic reticulum Ca(2+)-ATPase; TFEB: transcription factor; WTAP: WT1 associated protein; Wnt1: Wnt family member 1; YAP: Yes associated protein; YTHDF1/2: YTH N6-methyladenosine RNA binding protein F1/2.
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Zhang, H.; Lu, W.; Tang, H.; Chen, A.; Gao, X.; Zhu, C.; Zhang, J. Novel Insight of N6-Methyladenosine in Cardiovascular System. Medicina 2025, 61, 222. https://doi.org/10.3390/medicina61020222

AMA Style

Zhang H, Lu W, Tang H, Chen A, Gao X, Zhu C, Zhang J. Novel Insight of N6-Methyladenosine in Cardiovascular System. Medicina. 2025; 61(2):222. https://doi.org/10.3390/medicina61020222

Chicago/Turabian Style

Zhang, Huan, Wei Lu, Haoyue Tang, Aiqun Chen, Xiaofei Gao, Congfei Zhu, and Junjie Zhang. 2025. "Novel Insight of N6-Methyladenosine in Cardiovascular System" Medicina 61, no. 2: 222. https://doi.org/10.3390/medicina61020222

APA Style

Zhang, H., Lu, W., Tang, H., Chen, A., Gao, X., Zhu, C., & Zhang, J. (2025). Novel Insight of N6-Methyladenosine in Cardiovascular System. Medicina, 61(2), 222. https://doi.org/10.3390/medicina61020222

Article Metrics

Back to TopTop