Dilated cardiomyopathy (DCM) was the most prevalent cardiomyopathy worldwide. Although ferroptosis has been implicated in cardiac pathogenesis, its regulatory mechanism in DCM remained poorly defined. In this study, we found that GIPC1 (GAIP/RGS19-interacting protein), a scaffolding protein, was significantly downregulated in cardiac tissues from DCM patients and doxorubicin (DOX)-induced DCM models. Integrated proteomic and lipidomic analysis revealed that cardiac-specific knockout of GIPC1 disrupted mitochondrial fatty acid metabolism, increased the abundance of polyunsaturated fatty acid-containing phospholipids (PUFA-PLs), and ultimately promoted ferroptosis in cardiomyocytes. Both in vitro and in vivo experiments demonstrated that GIPC1 deficiency exacerbated ferroptosis and cardiac dysfunction in DOX-induced cardiomyopathy, whereas GIPC1 overexpression conferred protection against ferroptosis in DOX-induced cardiomyopathy. Mechanistically, co-immunoprecipitation mass spectrometry (Co-IP/MS) and molecular docking demonstrated that GIPC1 interacted with mitochondrial 2,4-dienoyl-CoA reductase (DECR1) via its PDZ domain. Surface plasmon resonance (SPR) analysis further confirmed a high-affinity direct binding between GIPC1 and DECR1 (KD = 16.3 nM). Co-IP and immunofluorescence (IF) demonstrated that GIPC1 facilitated actin-dependent transport of DECR1 into mitochondria, thereby maintaining redox homeostasis and suppressing ferroptosis. Consistently, DECR1 overexpression rescued GIPC1 ablation-induced ferroptosis by balancing redox homeostasis. Together, these results demonstrated that GIPC1 reduced cardiomyocyte susceptibility to ferroptosis by promoting mitochondrial translocation of DECR1 and remodeling lipid homeostasis, highlighting GIPC1/DECR1 axis as a potential therapeutic strategy for DCM. A schematic model illustrating the pathogenic cascade triggered by GIPC1 deficiency during DCM. In DCM, the expression level of GIPC1 was downregulated, thereby inhibiting actin-dependent transport of DECR1 into mitochondria, which remodeled lipid homeostasis and ultimately induced cardiomyocytes ferroptosis. Created with Figdraw.com.

BACKGROUND:Myocardial ischemia/reperfusion (I/R) injury induces an intense inflammatory response and involves multiple cell death pathways. PANoptosis, an integrated cell death process involving pyroptosis, apoptosis and necroptosis, is a major driver of cardiomyocyte loss during I/R injury. However, the epitranscriptomic control of PANoptosis is poorly understood. METHODS:We investigated the role of ALKBH3, an mRNA N1-methyladenosine (m1A) demethylase, in the regulation of cardiomyocyte PANoptosis using hypoxia/reoxygenation models in vitro and murine I/R models in vivo. Integrated transcriptomic and m1A epitranscriptomic profiling identified downstream targets. Loss- and gain-of-function studies of ALKBH3, AIM2, ZBED6 and STAT1 (siRNA or plasmid overexpression) were coupled with assessments of cell death phenotypes, inflammasome activity and gene expression. Molecular interactions and transcriptional/translational regulation were examined using co-immunoprecipitation, chromatin immunoprecipitation (ChIP) and dual-luciferase reporter assays. RESULTS:Cardiomyocyte-restricted ALKBH3 overexpression mitigates I/R injury in vivo. Mechanistically, ALKBH3 acts as a key suppressor of PANoptosis by inhibiting AIM2. ALKBH3 demethylates m1A onZBED6 mRNA, enhancing ZBED6 translation and limiting cardiomyocyte PANoptosis. Although ZBED6 does not bind directly to the AIM2 promoter, it physically interacts with STAT1, a transcriptional activator of AIM2, and represses STAT1-driven AIM2 expression. ZBED6 overexpression reduces AIM2 levels and PANoptosis, whereas AIM2 knockout attenuates the exacerbation of cardiac injury and PANoptosis induced by ALKBH3 silencing. CONCLUSIONS:These findings identify the ALKBH3/ZBED6/STAT1/AIM2 signalling axis that epitranscriptomically breaks cardiomyocyte PANoptosis, highlighting a tractable therapeutic target that limits cell death and improves myocardial outcomes after I/R.

Stem cell-derived extracellular vehicles (EVs) hold great therapeutic potential for myocardial infarction (MI). However, the efficient production of EVs with high bioactivity remains a critical bottleneck limiting their clinical translation. Here, we demonstrate that conditioned photobiomodulation (PBM) with green light is capable of activating human embryonic stem cells (hESCs) to secrete more EVs with superior cardioprotective activity. These PBM-reprogrammed hESC-EVs improve cardiac recovery in a murine MI model by promoting cardiomyocyte proliferation and angiogenesis while inhibiting apoptosis. Notably, we validate that these EVs similarly enhance the survival and proliferation of human cardiomyocytes, underscoring their translational potential. Further analysis reveals that this benefit is due to the higher miR-423-3p content in reprogrammed hESC-EVs, which enhances glycolytic metabolism and restores mitochondrial function by regulating the ZBTB7A/PKM2 axis. Moreover, we synthesize a methacryloyl hydrogel microneedle patch with superior biocompatibility, biodegradability, and mechanical strength for loading hESC-EVs, and convey the patch to the infarcted heart via a modified delivery apparatus. This system ensures the precise and sustained delivery of EVs to ischemic myocardium, offering a potent treatment for MI. Collectively, this optical and biomaterials-based approach efficiently prepares EVs with higher cardioprotective activity, providing new therapeutic strategies for heart disease.

During the healing phase after myocardial infarction (MI), various stimulatory factors activate the transformation of cardiac fibroblasts (CFs) into myofibroblasts, leading to collagen synthesis and cardiac fibrosis. Here, we investigated the role of the cardiac ischemia‒reperfusion-associated Ku70-interacting long noncoding RNA (CIRKIL) in post-MI cardiac fibrosis and the underlying molecular mechanisms. Cardiac fibrotic tissue from MI mice and transforming growth factor-β (TGF-β1)-treated CFs exhibited elevated expression of CIRKIL in the cytosol of the CFs. Furthermore, CIRKIL gain-of-function exacerbated MI-induced cardiac fibrosis in transgenic mice, whereas CIRKIL deficiency alleviated this fibrotic progression. Consistently, CIRKIL overexpression promoted the proliferation and transformation of CFs, resulting in the upregulation of collagen type I and type III, whereas CIRKIL silencing had the opposite effects. Mechanistically, CIRKIL functioned as a molecular scaffold that facilitated the functional oligomerization of the pyruvate kinase M2 (PKM2) homotetramer, which binds to Smad7 and inhibits Smad7-mediated ubiquitination and degradation of TGF-β type I receptor (TGFBR1), a critical profibrotic factor in the TGF-β signaling pathway. Importantly, the regulatory effects of the functional fragment of CIRKIL on PKM2 homotetramer formation and CF activation were similar to those of full-length CIRKIL. Our findings revealed that CIRKIL, as a molecular scaffold, facilitates the assembly of the PKM2 homotetramer, which inhibits the Smad7-mediated ubiquitination and degradation of TGFBR1 and results in CF activation and cardiac fibrosis in mouse MI hearts, indicating that CIRKIL might be a promising intervention target for cardiac fibrosis-related diseases.

Objective:Myocardial ischemia-reperfusion(I/R)injury remains a major contributor to cardiac morbidity and mortality,and accumulating evidence suggests that epitranscriptomic regulation may critically influence cardiac stress responses.N6-methyladenosine(m6A)modification and circular RNAs(circRNAs)have emerged as important regulators of cardiovascular pathology;however,their integrated roles in myocardial I/R injury,particularly under chronic cold stress,remain poorly defined.Methods:A mouse model of myocardial I/R injury was established under room-temperature or chronic cold exposure conditions.Cardiac function,infarct size,histopathology,and serum injury markers were assessed.Global m6A levels were quantified,and m6A-modified circRNA profiles were analyzed using epitranscriptomic microarrays and bioinformatics approaches.Differentially expressed circRNAs were validated in vivo and in hypoxia-reoxygenation-treated neonatal cardiomyocytes.Circular structures and stability were confirmed by Sanger sequencing,divergent/convergent PCR,and actinomycin D assays.Competing endogenous RNA(ceRNA)networks were constructed to identify downstream regulatory pathways.Results:Myocardial I/R injury resulted in significant cardiac dysfunction,increased infarct size,histological damage,and elevated serum CK-MB and LDH levels,accompanied by a marked increase in global m6A methylation.Epitranscriptomic profiling identified 391 circRNAs with altered m6A modification following I/R injury,involving pathways related to molecular binding,cellular processes,and kinase signaling.Multiple circRNAs exhibited consistent dysregulation in both in vivo and in vitro I/R models and displayed high structural stability.Importantly,chronic cold exposure significantly exacerbated I/R-induced cardiac dysfunction and infarct severity while further modulating the expression of specific m6A-modified circRNAs.ceRNA network analysis revealed that cold-responsive circRNAs potentially regulate myocardial injury through miRNA-mediated signaling pathways.Conclusion:This study identifies m6A-modified circRNAs as key epitranscriptomic regulators of myocardial I/R injury and demonstrates that chronic cold stress amplifies circRNA-mediated regulatory networks.These findings provide novel mechanistic insight into temperature-dependent epigenetic regulation in ischemic heart disease and highlight m6A-circRNAs as potential therapeutic targets.

Cardiac aging is associated with progressive cardiac fibrosis and dysfunction, yet the underlying mechanisms remain incompletely understood. Extrachromosomal circular DNA (eccDNA) has been reported to participate in tumor and age-related genomic instability, while its role in cardiac fibrosis during aging remains to be fully elucidated. In this study, circular DNA sequencing and RNA seqencing were performed to analyze eccDNA profiles in young and aged cardiac tissues. The number of eccDNAs in the cardiac tissue of aged mice is higher than that in young mice. Combining the annotation of eccDNAs and the key genes related to aging identified in the transcriptome, we identified sterile alpha and TIR motif containing 1 (Sarm1), a key regulator of NAD+ metabolism and neurodegeneration located in eccDNAs, as a novel driver of cardiac aging via pro-fibrotic signaling. In aged mice, Sarm1 knockdown significantly restored cardiac function and reduced fibrosis. Conversely, Sarm1 accelerated cardiac aging phenotypes in young Sarm1-overexpressing transgenic mice. Mechanistically, co-immunoprecipitation combined with mass spectrometry identified TGF-β-Smad2/3 as the dominant pathway, with pharmacological inhibition by SIS3 abolishing Sarm1-driven Smad2/3 phosphorylation. Our findings reveal that Sarm1-containing eccDNA drives cardiac aging by amplifying pro-fibrotic signaling through the TGF-β-Smad2/3 pathway, proposing eccDNAs clearance and Sarm1 inhibition as novel therapeutic strategies for aging-related cardiac fibrosis.

The mechanisms that define the metabolic capacity of the heart remain incompletely understood. Here, we identify translational capacity as a rate-limiting determinant of cardiac metabolic output, mediated by a ribosome–mitochondria coupling axis linking cytosolic and mitochondrial protein synthesis. Integrated transcriptomic and proteomic analyses reveal extensive discordance between mRNA and protein abundance, accompanied by suppression of global protein synthesis, reduced ribosome biogenesis, and disruption of proteostasis. We identify a ribosome–mitochondria coupling axis in which the cytosolic ribosomal protein RPS5 regulates the abundance and mitochondrial localization of MRPS5, a component of the mitochondrial ribosome. This process occurs in association with endoplasmic reticulum–mitochondria contact sites and links cytosolic translation to mitochondrial protein synthesis. Perturbation of this axis is associated with reduced expression of metabolic enzymes and impaired mitochondrial function. Conversely, restoration of RPS5 or MRPS5 improves mitochondrial activity and cardiac performance in vivo. These findings suggest that translational capacity contributes to metabolic homeostasis and reveal a spatially organized mechanism linking protein synthesis to mitochondrial function in the diabetic heart.

Stem cell therapies are emerging as promising strategies for repair after myocardial infarction (MI), but the repair efficacy is limited by the poor cardiac microenvironment represented by the inflammatory response, as well as oxidative stress, and adverse electrical coupling. Here, we developed an injectable supramolecular hydrogel (HCPA) that modulates the infarct microenvironment and accelerates myocardial repair by encapsulating human induced pluripotent stem cells derived cardiomyocytes (hiPSC-CMs). HCPA hydrogel not only exhibited excellent reactive oxygen species (ROS) response in order to minimize oxidative stress but also possessed desirable electrical conductivity for the reintegration of electrical impulses. Critically, RNA sequencing demonstrated that the PPARα/NFκB pathway contributed significantly to the HCPA hydrogel-promoted macrophage polarization from M1-type to M2-type, thus alleviating inflammatory responses. HCPA hydrogel harboring hiPSC-CMs increased retention of hiPSC-CMs and improved cardiac function in MI mice. This study represents a new integrated therapeutic option for MI and provides insights for the development of novel biomaterials in the field of tissue engineering.

Cardiac ischemia‒reperfusion (I/R) injury is a leading cause of disability and mortality worldwide, but the underlying mechanism remains largely unknown. Despite the emerging recognition of circular RNAs (circRNAs) as pivotal regulators of cardiac development and disease, their roles in cardiac I/R injury have yet to be thoroughly investigated. In this study, we identified a circRNA named circArhgap26, which is regulated by m6A modification. The expression of circArhgap26 was significantly decreased in the I/R myocardium. Cardiac-specific overexpression of circArhgap26 ameliorated cardiac dysfunction and reduced the infarct area and cardiomyocyte apoptosis in I/R model mice. Mechanistically, circArhgap26 directly bound to PKP1, thereby inhibiting the interaction between PKP1 and the palmitoyltransferase ZDHHC1. The subsequent palmitoylation of PKP1 and its protein stability are subsequently diminished, leading to a reduction in APAF1 protein synthesis and the inhibition of the Caspase-9/Caspase-3 signaling pathway, thereby mitigating cardiomyocyte apoptosis. Most importantly, the expression of circArhgap26 in the plasma of patients undergoing percutaneous coronary intervention (PCI) was decreased. This study not only elucidates the dual regulatory mechanisms of circArhgap26, m6A modification and posttranslational modification (palmitoylation), in combating I/R injury but also provides a theoretical foundation for circRNA-based therapies. Its dual value as a prognostic biomarker and therapeutic target holds promise for advancing precision cardiovascular medicine and improving outcomes in globally prevalent I/R-related diseases.

Coronary heart disease (CHD), characterized by impaired coronary artery function, often results in myocardial ischemia, hypoxia, and necrosis, with clinical manifestations such as angina pectoris. Vascular smooth muscle cell (VSMC) hypercontraction, primarily regulated by intracellular calcium (Ca2+), plays a central role in pathological coronary vasoconstriction. This study aimed to evaluate the therapeutic effects of Huatuo Zaizao Pills (HTZZ) in alleviating myocardial ischemia caused by abnormal coronary artery contraction and to elucidate the underlying molecular mechanisms. A double-blind, multicenter, randomized, placebo-controlled clinical trial was conducted to assess the efficacy of HTZZ in patients with angina pectoris. In vivo, pituitrin-induced acute myocardial ischemia mice and spontaneously hypertensive rats (SHRs) were used to evaluate myocardial and vascular responses to HTZZ. In vitro, vasorelaxation mechanisms were investigated using isolated rat mesenteric arterial rings, patch clamp, calcium imaging, and [3H]-ryanodine binding assays. HTZZ significantly reduced the frequency and duration of angina attacks in clinical settings. It improved myocardial ischemia in mice and enhanced vascular elasticity and diastolic function in SHRs. Mechanistically, HTZZ induced vasodilation by inhibiting extracellular Ca2+ influx and reducing intracellular Ca2+ levels via suppression of L-type calcium channels (LTCCs) and ryanodine receptors (RyRs). Long-term HTZZ administration also downregulated LTCC expression at both the protein and mRNA levels. HTZZ effectively alleviates angina and myocardial ischemia by suppressing Ca2+-mediated vasoconstriction through targeting LTCCs and RyRs. These findings highlight HTZZ as a promising therapeutic candidate for CHD characterized by coronary vasospasm and ischemia.

Cardiac fibroblasts progressively replace deceased cardiomyocytes during the development of myocardial fibrosis, an irreversible pathological repair process that ultimately leads to cardiac dysfunction and heart failure. Cardiac injury was evaluated by echocardiography and Masson staining in myocardial infarction (MI) mice with zinc finger BED-type containing 6 (ZBED6) knockdown or overexpression. Furthermore, chromatin immunoprecipitation (ChIP) assays, electrophoretic mobility shift assays (EMSAs), and luciferase reporter assays were used to explore the target of ZBED6. ZBED6 expression was notably decreased in vivo in MI hearts and in vitro in TGF-β-induced primary mouse cardiac fibroblasts (PMCFs). Transgenic overexpression of ZBED6 specifically in cardiac fibroblasts improved cardiac dysfunction, reduced the infarct area, and decreased the expression levels of fibrotic genes after MI injury. Conversely, physiological knockdown of ZBED6 induced cardiac dysfunction and remodeling, which is consistent with the phenomena observed in vitro. Mechanistically, ZBED6, which functions as a transcriptional inhibitor of Piezo1, failed to prevent its transcription owing to mutations in the promoter binding sites. Stimulation of Piezo1 in PMCFs facilitates YAP translocation into the nucleus, whereas knockdown of Piezo1 or the use of a Piezo1 inhibitor suppresses this translocation. Moreover, the activation of Piezo1 reversed the cardioprotective effects of ZBED6 overexpression. In summary, the protective effect of ZBED6 against myocardial fibrosis injury is achieved through the inhibition of Piezo1 transcription, leading to reduced YAP nuclear translocation. These findings suggest that ZBED6 may become a potential therapeutic target for the clinical treatment of myocardial fibrosis.

Heart failure with reduced ejection fraction (HFrEF) remains a major therapeutic challenge. B-cell lymphoma 2-associated transcription factor 1 (Bclaf1) is implicated in RNA splicing and cardiac disease, but its role in HFrEF pathogenesis is unknown. Here, we demonstrate that Bclaf1 expression is elevated in human HFrEF myocardium and in male murine pressure-overload models. Cardiac-specific Bclaf1 overexpression drives pathological hypertrophy and systolic dysfunction, whereas its genetic knockout or adeno-associated virus serotype 9 (AAV9)-mediated knockdown attenuates these phenotypes. Mechanistically, Bclaf1 interacts with the splicing factor serine/arginine-rich splicing factor 2 (Srsf2) to bind to heart and neural crest derivatives expressed 2 (Hand2) pre-mRNA and enhance its splicing efficiency, leading to increased mature Hand2 levels and maladaptive remodeling. Inhibition of either Bclaf1 or Hand2 rescues cardiac function and structure in experimental HFrEF. Our work defines a Bclaf1/Srsf2/Hand2 splicing axis as a critical driver of HFrEF and reveals a promising therapeutic target for heart failure.

PURPOSE:Recent research indicates that the senescence of bone marrow mesenchymal stem cells (BMSCs) disrupts the osteo-adipogenic balance, a primary factor contributing to the development of osteoporosis. Our previous findings have implicated methyltransferases in this process, among which methyltransferase-like 13 (METTL13) has been established to regulate cell fate, although its role in osteoporosis has yet to be determined. METHODS:Bone formation was assessed using micro-computed tomography and hematoxylin and eosin staining. Protein expression in bone tissues was examined immunohistochemically, and cellular mRNA and protein levels were determined using quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and western blotting. Cellular senescence was evaluated based on β-galactosidase staining, and osteogenic and adipogenic differentiation was examined using alkaline phosphatase, Alizarin Red S, and Oil Red O staining. Protein interactions and DNA binding were determined using co-immunoprecipitation and chromatin immunoprecipitation. RESULTS:METTL13 expression was significantly enhanced in ovariectomy-induced senescent bone and BMSCs, whereas METTL13 knockdown markedly reversed etoposide-induced cellular senescence. By binding to forkhead box protein A1 (Foxa1), METTL13 promotes the preferential differentiation of BMSCs into adipocytes, as opposed to osteocytes. Moreover, Foxa1 had effects opposite to those of METTL13 on BMSC differentiation, inhibiting the nuclear entry of METTL13. Notably, blocking the nuclear import of Foxa1 suppressed the transcriptional expression of HES-1, which promoted the adipogenic differentiation of BMSCs and inhibited osteogenic differentiation. CONCLUSIONS:Our findings in this study revealed the mechanisms whereby METTL13 promotes BMSC senescence and disrupts BMSC differentiation, on the basis of which, we identified the METTL13-Foxa1-HES-1 axis as a potential therapeutic target for treatment of osteoporosis.

Previous studies have highlighted the significance of RNA-binding proteins and alternative splicing (AS) in the progression of complex diseases, but the specific involvement of AS in heart failure (HF) remains unclear. This study aimed to elucidate the role of RNA-binding motif single-stranded interacting protein 1 (RBMS1), an RNA-binding protein, in the development of HF by regulating AS and its effect on cardiac fibrosis. The level of RBMS1 was investigated in the hearts of both HF patients and mice. Fibroblast-specific knockout RBMS1 mice were generated to investigate the role of RBMS1 in cardiac fibrosis and HF. Unbiased RNA sequencing and RNA immunoprecipitation combined with RNA pull-down were conducted to identify the downstream effector of RBMS1 in fibroblasts. RBMS1 expression was increased in murine hearts following myocardial infarction, as well as in the hearts of patients with ischaemic cardiomyopathy and hypertrophic cardiomyopathy. Moreover, RBMS1 levels in the hearts of HF patients were positively associated with cardiac fibrosis. Furthermore, fibroblast-specific ablation of RBMS1 improved cardiac dysfunction by mitigating myocardial fibrosis. Mechanistically, RBMS1 regulated the alternative splicing of LIM domain 7 (LMO7) by binding to intron 19 and splicing out exon 20, resulting in the formation of the LMO7-Δe20 isoform, which thus activated the transforming growth factor (TGF)-β1 pathway by upregulating activator protein 1. More importantly, overexpression of LMO7-Δe20 in mice resulted in cardiac fibrosis and cardiac dysfunction, which was ablated after treatment with TGF-β1 pathway inhibitor SB431542. In addition, SB431542 attenuated the RBMS1-driven fibrogenesis in human cardiac fibroblasts. Strikingly, pharmacologically inhibiting RBMS1 by low-dose nortriptyline or antisense oligonucleotide-mediated RBMS1 deficiency alleviated myocardial fibrosis and improved cardiac function in HF mice. These findings unveil a critical role of RBMS1 in regulating cardiac fibrosis through controlling the splicing of LMO7 to activate the TGF-β1 pathway. Genetic ablation or pharmacological inhibition of RBMS1 improves cardiac function in mice, suggesting its potential as a therapeutic target for HF.

Diabetic cardiomyopathy (DCM) is a medical condition characterized by cardiac remodeling and dysfunction in individuals with diabetes mellitus. Sarcoplasmic reticulum (SR) and mitochondrial Ca2+ overload in cardiomyocytes have been recognized as biological hallmarks in DCM; however, the specific factors underlying these abnormalities remain largely unknown. In this study, we aimed to investigate the role of a cardiac-specific long noncoding RNA, D830005E20Rik (Trdn-as), in DCM. Our results revealed the remarkably upregulation of Trdn-as in the hearts of the DCM mice and cardiomyocytes treated with high glucose (HG). Knocking down Trdn-as in cardiac tissues significantly improved cardiac dysfunction and remodeling in the DCM mice. Conversely, Trdn-as overexpression resulted in cardiac damage resembling that observed in the DCM mice. At the cellular level, Trdn-as induced Ca2+ overload in the SR and mitochondria, leading to mitochondrial dysfunction. RNA-seq and bioinformatics analyses identified calsequestrin 2 (Casq2), a primary calcium-binding protein in the junctional SR, as a potential target of Trdn-as. Further investigations revealed that Trdn-as facilitated the recruitment of METTL14 to the Casq2 mRNA, thereby enhancing the m6A modification of Casq2. This modification increased the stability of Casq2 mRNA and subsequently led to increased protein expression. When Casq2 was knocked down, the promoting effects of Trdn-as on Ca2+ overload and mitochondrial damage were mitigated. These findings provide valuable insights into the pathogenesis of DCM and suggest Trdn-as as a potential therapeutic target for this condition.

Cold exposure activates brown adipose tissue (BAT), to alleviate metabolic disorders. However, the mechanisms underlying the regulation of mitochondrial lipid droplet contact (MLC) in BAT and their association with these benefits remain unclear. Here, we identify liver-derived β-hydroxybutyrate (BHB) as a key mediator in driving MLC formation in BAT. Mechanistically, BHB directly targets at the GLY-67 residue of RAB10, enhancing its interaction with PLIN5 to form the RAB10-PLIN5 complex, which facilitates MLC. This interaction was validated using SPIDER and biotin-labeled pull-down assays. Functionally, BHB treatment reduces lipotoxicity and improves metabolic health in diet-induced obese mice. These findings establish BHB as a critical link between BAT MLC and the systemic metabolic benefits, highlighting the RAB10-PLIN5 complex as a therapeutic target for obesity and hepatic steatosis. Furthermore, this work underscores the broader significance of cold-induced metabolic adaptations for combating metabolic diseases.

Liver cancer is a highly aggressive malignancy with poor survival rates. Current treatments, including liver transplantation, immunotherapy, and gene therapy, are often limited by late-stage diagnosis and significant side effects, highlighting the urgent need for novel therapeutic agents. In this study, we evaluated the therapeutic potential of Kanglexin (KLX), a novel anthraquinone derivative, in the treatment of liver cancer. In vitro, KLX inhibited the proliferation and migration of HepG2 and Hep3B cells in a dose-dependent manner. Mechanistically, KLX upregulated Z-DNA binding protein 1 (ZBP1) expression, inducing PANoptosis by directly binding to ZBP1, altering its conformation, and reducing its affinity for the E3 ubiquitin ligase ring finger protein 180 (RNF180). This interaction decreased ZBP1 ubiquitination, thereby increasing its stability. Additionally, KLX upregulated the expression of the transcription factor homeobox D10 (HOXD10), which further increased ZBP1 expression. Elevated ZBP1 levels significantly suppressed liver cancer cell proliferation and migration, whereas the inhibitory effects of KLX were reversed upon ZBP1 knockdown. In a xenograft model, KLX significantly inhibited tumor growth with a lower toxicity than oxaliplatin (OXA). In conclusion, KLX promoted PANoptosis in liver cancer cells by upregulating ZBP1 and preventing its degradation, thereby inhibiting liver cancer progression and migration. These findings suggest that KLX is a promising therapeutic agent for liver cancer.

Cardiovascular disease remains the foremost contributor to mortality and disability globally, even with significant advancements in prevention, diagnosis, and early intervention. A comprehensive insight into cardiovascular diseases and the intrinsic molecular mechanisms is critical to innovating more effective therapeutic interventions for prevention and therapy. Latest advancements within the realm of epigenetic modulation, especially methylation modification, of gene expression have corroborated the impacts of epigenetic modifications in governing the pathogenesis and progression of cardiovascular diseases and suggested the viability of epigenetic mechanisms as emerging targets for the development of new diagnostic and therapeutic strategies. In this review, we first provide a brief overview of the biological processes of methylation modifications, including DNA methylation, protein methylation, and RNA N6-methyladenosine (m6A) modification. We then summarize their roles in cardiac hypertrophy, heart failure, ischemic heart disease, and atherosclerosis.

Dystrophin is a critical interacting protein of Nav1.5 that determines its membrane anchoring in cardiomyocytes. Long noncoding RNAs (lncRNAs) are involved in the regulation of cardiac ion channels, while their influence on sodium channel remains unexplored. Our preliminary data showed that lncRNA-Dachshund homolog 1 (lncDACH1) can bind to dystrophin, which drove us to investigate if lncDACH1 can regulate sodium channel by interfering with dystrophin. Western blot and immunofluorescent staining showed that cardiomyocyte-specific transgenic overexpression of lncDACH1(lncDACH1-TG) reduced the membrane distribution of dystrophin and Nav1.5 in cardiomyocytes. Meanwhile, peak I Na were reduced in the hearts of lncDACH1-TG mice than wild-type (WT) controls. The opposite data of western blot ,immunofluorescent staining and patch clamp were collected from lncDACH1 cardiomyocyte conditional knockout (lncDACH1-cKO) mice. Moreover, increased ventricular arrhythmia susceptibility was observed in lncDACH1-TG mice in vivo and ex vivo . The conservative fragment of lncDACH1 inhibited membrane distribution of dystrophin and Nav1.5, and promoted the inducibility of ventricular arrhythmia. Strikingly, activation of dystrophin transcription by dCas9-SAM system in lncDACH1-TG mice rescued the impaired membrane distribution of dystrophin and Nav1.5, and prevented the occurrence of ventricular arrhythmia. Furthermore, lncDACH1 was increased in transaortic constriction (TAC) induced failing hearts, which promoted the inducibility of ventricular arrhythmia. And the expression of lncDACH1 is regulated by hydroxyacyl-CoA dehydrogenase subunit beta (hadhb), which binds to lncDACH1 and decreases its stability. The human homologue of lncDACH1 inhibited the membrane distribution of Nav1.5 in human iPS-differentiated cardiomyocytes. The findings provide novel insights into the mechanism of Nav1.5 membrane targeting and the development of ventricular arrhythmias.