BACKGROUND & AIMS:Increased de novo lipogenesis, largely mediated by sterol regulatory element-binding protein 1 (SREBP1), is a hallmark of metabolic dysfunction-associated steatotic liver disease (MASLD). However, the post-translational mechanisms regulating SREBP1 turnover remain poorly understood. The endoplasmic reticulum-associated degradation (ERAD) pathway ensures protein quality and quantity control, yet its role in hepatic lipid metabolism remains elusive. Here, we investigate the function of the ERAD-specific E3 ubiquitin ligase MARCHF6 in hepatic lipid homeostasis and MASLD pathogenesis. METHODS:Liver-specific Marchf6 knockout (Marchf6Alb) mouse cell lines and organoids were generated to assess the impact of Marchf6 depletion on hepatic lipid metabolism under normal chow, high-fat diet, and Western diet conditions. RNA sequencing, proteomics, biochemical and molecular biological analyses were performed to identify molecular pathways regulated by MARCHF6. Functional studies in primary hepatocytes, human hepatoma cells and organoids were conducted to determine the mechanistic link between MARCHF6 and SREBP1. RESULTS:Hepatic MARCHF6 expression was significantly reduced in both MASLD mouse models and human patients. Liver-specific Marchf6 deletion aggravated hepatic lipid accumulation, fibrosis, and inflammation. Transcriptomic and proteomic analyses revealed upregulation of lipogenic genes in Marchf6Alb livers, with a marked increase in SREBP1 protein levels. Mechanistically, MARCHF6 directly interacted with and ubiquitinated SREBP1, targeting it for proteasomal degradation. Loss of MARCHF6 prolonged SREBP1 half-life, driving excessive de novo lipogenesis. CONCLUSIONS:MARCHF6-ERAD is a critical regulator of hepatic lipid metabolism, functioning as a sterol binding protein to control SREBP1 turnover. Its downregulation promotes hepatic steatosis and MASLD progression, highlighting MARCHF6 as a potential therapeutic target in MASLD. IMPACT AND IMPLICATIONS:This study identifies the endoplasmic reticulum-resident E3 ubiquitin ligase MARCHF6 as a key regulator of SREBP1 stability, hepatic lipid homeostasis and MASLD (metabolic dysfunction-associated steatotic liver disease) progression. We demonstrate that loss of MARCHF6 promotes hepatic steatosis and fibrosis, whereas restoration of MARCHF6 largely reverses these phenotypes, highlighting a reversible and therapeutically targetable pathway. These findings provide new mechanistic insight into lipid dysregulation in MASLD and position the MARCHF6-SREBP1 axis as a promising target for metabolic liver disease intervention.

Androgen excess is a common feature of polycystic ovary syndrome, congenital adrenal hyperplasia, gender affirming hormone therapy and is a known driver of disrupted glucose homeostasis. However, its impact on hepatic lipid regulation remains poorly understood with conflicting results. In age and weight matched female mice, dihydrotestosterone (DHT) treatment under a regular chow diet did not alter hepatic or circulating lipid levels. In contrast, under a western diet, DHT protected against metabolic dysfunction-associated steatotic liver disease by reducing hepatic triglyceride (TG) accumulation, even as it exacerbated systemic hypertriglyceridemia. These effects occurred without changes in hepatic insulin-Akt signaling and were independent of hepatic androgen receptor signaling. Multiomics profiling revealed that DHT reprogrammed hepatic transcription toward a male like pattern driven by pulsatile growth hormone (GH)-STAT5 signaling. DHT enhanced hepatic STAT5 activation, suppressed expression of the fatty acid (FA) transporter Cd36, and reduced hepatic FA uptake, even in the presence of elevated circulating free fatty acid. DHT also increased VLDL-TG secretion and altered hepatic FA composition. Continuous GH infusion, which mimics the female GH secretion pattern, reversed these effects by attenuating STAT5 activation and restoring Cd36 expression. Together, these findings identify a GH-dependent, hepatic androgen receptor-independent pathway through which androgen excess shapes hepatic lipid homeostasis. They further show that DHT elicits divergent systemic effects, with impaired glucose homeostasis and enhanced lipolysis occurring alongside reduced hepatic lipid accumulation but increased TG secretion.

Signal transducer and activator of transcription 2 (STAT2) is a key component of the type I interferon (IFN-I/III) signaling pathway, which is pivotal in host defense against cancer and viral infections and in shaping immune responses. Building on our previously reported conditional Stat2 knockout (KO) mouse, we expand its utility by validating additional tissue-specific models and exploring novel functional contexts. Mice carrying loxP-flanked Stat2 alleles were crossed with CMV-Cre, Cdx2-Cre or CD11c-Cre mice. Deletion of STAT2 was validated by PCR genotyping and western blotting in the relevant tissues. To confirm defective IFN-I signaling with STAT2 deletion, IFN-β stimulation of splenocytes from CMV-Cre Stat2 KO mice showed a lack of induction of canonical IFN-I target genes, confirming functional disruption of the pathway. In vivo, global Stat2 deletion significantly impaired the antitumor efficacy of IFN-β treatment. Similarly, lung fibroblasts isolated from globally deleted Stat2 KO mice showed defective antiviral responses to IFN-β. Tissue-specific Cre models demonstrated selective ablation of STAT2 in target compartments without affecting its expression in non-target tissues. Together, these studies expand our published conditional Stat2 KO findings and highlight the value of this model as a versatile platform for dissecting STAT2-dependent signaling pathways in a tissue- and disease-specific manner.

Vascular tissues provide long-distance transport and physical support in the vascular plant lineage, providing a significant adaptive advantage. Although the cross talk between auxin and cytokinin in promoting both vascular cell proliferation and differentiation has been well studied in angiosperms such as Arabidopsis thaliana , little is known about this regulation in other vascular plant lineages. Here, we found that unlike the hormonal cross talk found in all other species under study, the lycophyte Selaginella moellendorffii shows clear task separation, with auxin driving vascular cell proliferation only and cytokinin specifically triggering cell differentiation. Using a cross-species transcriptomics approach, we found that members of the AUXIN / INDOLE-ACETIC ACID ( AUX / IAA ) and CYTOKININ OXIDASE ( CKX ) gene families exhibited divergent expression patterns in response to auxin and cytokinin treatments. Despite these regulatory differences, we show that AUX/IAA and CKX proteins are functionally conserved between Arabidopsis and Selaginella. Taken together, our findings suggest an evolutionary adaptation to the hormonal regulation of vascular tissue development in which core protein functions are conserved, but regulatory circuits diverged in lycophytes.

We are delighted to share with you our fifteenth Journal Club and highlight some of the most interesting papers published recently [...].

Escherichia coli (E. coli) is a leading cause of invasive bacterial infections in humans. Pathogenic E. coli is not only the major etiological agent of enteric/diarrheal disease and urinary tract infections, but also among the most common causes of sepsis and meningitis. Caspase-8 is known to regulate apoptotic and pyroptotic cell death in response to bacterial and viral infections. Here we demonstrate that caspase-8 plays a critical role in E. coli-induced macrophage apoptosis in vitro and in regulating immune response and host death in vivo. Incubation of mouse bone marrow derived macrophages (BMDMs) with an E. coli K1 strain CE10 triggered robust cell death, which is independent of the NAIP/NLRC4/caspase-1/GSDMD pathway. CE10 stimulation induced caspase-8 activation, and macrophages deficient in caspase-8 and RIPK3, but not RIPK3 alone, were protected from CE10-induced cell death. In an intraperitoneal injection sepsis model, E. coli-induced IL-1β, TNF-α, and IL-6 production was markedly reduced in caspase-8-/-/RIPK3-/- mice, compared with RIPK3-/- or wild type mice. Accordingly, the survival rate was significantly improved in caspase-8-/-/RIPK3-/- mice. Moreover, caspase-8 deficiency attenuated CE10-induced NF-κB activation and cytokine production in BMDMs. Together, our findings identify caspase-8 as a central mediator of E. coli-induced cell death, immune response, and establish its critical contribution to host mortality during E. coli infection.

Pyroptosis is a form of proinflammatory cell death characterized by inflammasome activation, pore formation, and the release of pro-inflammatory cytokines such as interleukin-1β (IL-1β) and IL-18 upon cell rupture. Nuclear factor-κB (NF-κB), a prototypical pro-inflammatory transcription factor, plays a critical role in immune system regulation. Recent research highlights the multifaceted roles of NF-κB signaling in pyroptosis. Various immunologically relevant ligands and their receptors can activate the NF-κB pathway to promote pyroptosis, with Toll-like receptors (TLRs), IL-1 receptors (IL-1Rs), and TNF receptors (TNFRs) being the most prominent. NF-κB regulates the transcription of key components of inflammasomes involved in pyroptosis, particularly the NLRP3 inflammasome. Recent studies also indicate that NF-κB modulates the activation of NLRC4 and AIM2 inflammasomes through distinct pathways in diverse inflammatory conditions, such as acute lung injury and neuroinflammation. Additionally, the NF-κB pathway mediates the production of inflammatory cytokines, including IL-1β, IL-33, and TNF-α, which further regulate pyroptosis. This review examines recent advances in understanding the role of the NF-κB signaling pathway in regulating pyroptosis during infection and inflammation.

The Salmonella enterica serovar Typhimurium (ST) mutant lacking the msbB gene (ΔmsbB) has been widely studied as a candidate for attenuated bacterial vectors in therapeutic applications. Deletion of msbB results in LPS with under-acylated lipid A, which lowers endotoxicity while maintaining structural integrity. This attenuation has traditionally been attributed to reduced TLR4 activation due to weaker interaction between the modified lipid A and TLR4. In our study, we confirmed that ΔmsbB ST was less lethal than wild-type (WT) ST in a mouse sepsis model. However, this difference persisted even in TLR4- and caspase-11-deficient mice, suggesting that LPS signaling is not the primary determinant of virulence. In vitro, bone marrow–derived macrophages (BMDMs) from TLR4- or caspase-11-deficient mice showed only modest reductions in ST-induced cell death and cytokine production. Importantly, ΔmsbB ST behaved similarly to WT ST in these assays, further indicating that LPS-mediated signaling is not central to the observed attenuation. Our previous studies showed that ST-induced mortality in mice is primarily mediated through NLRC4 activation. Using qPCR and immunoblotting, we found that expression of NLRC4 activators was diminished in the ΔmsbB strain. Additionally, the mutant exhibited increased outer membrane permeability—likely contributing to its heightened antibiotic sensitivity—and reduced motility due to lower flagellin protein levels. In summary, the attenuation of virulence observed in the ΔmsbB strain is not directly due to altered LPS–TLR4 interactions, but rather an indirect effect of diminished expression of virulence factors that activate the NLRC4 inflammasome.

The LSM (Like-Smith) family comprises RNA-binding proteins (RBPs) that are key regulators of RNA metabolism. LSM14A, a member of this family (designated Lsm14a in mice), participates in RNA processing within cytoplasmic processing bodies (P-bodies). The mouse Lsm14a gene is localized to chromosome 7qB1, spans 48.67 kilobases (kb), and encodes a 462-amino-acid protein that exhibits 94.53 % amino acid identity with human LSM14A. However, the expression profile of LSM14A in male reproductive organs and its functional relevance to male fertility remain uncharacterized. In this study, we report that LSM14A is expressed in the mouse testis and localizes to the cytoplasm of germ cells, from spermatogonia to elongating spermatids. To investigate LSM14A function, we generated germ cell-specific Lsm14a conditional knockout (cKO) mice. Lsm14a cKO male mice displayed normal growth, development, and fertility. Histological examination of Lsm14a cKO testes revealed preserved spermatogenesis and seminiferous tubule structure. Lsm14a cKO sperm exhibited normal morphology, acrosome integrity, and motility. The loss of Lsm14a in the testes did not significantly affect P-body formation, suggesting that genetic compensation by other LSM family members may have been activated upon Lsm14a knockout, thereby compensating for its loss of function. Collectively, these findings demonstrate that LSM14A is dispensable for spermatogenesis and male fertility in mice.

Macrophages play a crucial role in both innate immunity and inflammation. The NAIP/NLRC4 inflammasome is an immune sensor that detects virulence factors from invasive Gram-negative bacteria, including flagellin and type III secretion system (T3SS) proteins, to trigger pyroptotic cell death. Mitochondria are emerging as important regulators of inflammasome signaling; however, their contribution to NLRC4 activation remains unclear. Here, we show that mitochondria serve as essential platforms for NAIP/NLRC4 inflammasome assembly and activation in macrophages. Mitochondrial depletion in mouse immortalized bone marrow-derived macrophages (iBMDMs) significantly impairs ASC speck formation and NLRC4 inflammasome activation. NLRC4 activation in mouse BMDMs lacking mitochondrial transcription factor A (TFAM) was also diminished. Upon activation by Salmonella or EprJ (a rod protein of E. coli T3SS), NLRC4, ASC, and Caspase-1 are recruited to mitochondria in a time-dependent manner in mouse BMDMs. NLRC4 activation subsequently induces mitochondrial dysfunction through Caspase-1- and Caspase-8-mediated cleavage of the pro-apoptotic factor BID, serving as a positive feedback loop between mitochondrial damage and inflammasome signaling that further amplifies the inflammasome response. Our findings reveal a critical interplay between mitochondria and inflammasomes, providing new insights into host–pathogen interactions and suggesting potential strategies for targeting inflammasome-related diseases.

IntroductionKlebsiella pneumoniae is a Gram-negative bacterium and the third most commonly isolated microorganism in blood cultures from septic patients. Despite extensive research, the mechanisms underlying K. pneumoniae-induced sepsis and its pathogenesis remain unclear. Acute respiratory failure is a leading cause of mortality in systemic K. pneumoniae infections, highlighting the need to better understand the host immune response and bacterial clearance mechanisms.MethodTo investigate the impact of K. pneumoniae infection on organ function and immune response, we utilized a systemic infection model through intraperitoneal injection in mice. Bacterial loads in key organs were quantified, and lung injury was assessed. Survival analysis was performed in wild-type (WT) and gene deficient mice. Mitochondrial damage and reactive oxygen species (ROS) production, as well as cytokine levels were measured in macrophages isolated from these mice to evaluate their contribution to bacterial clearance capacity.ResultsOur findings demonstrate that K. pneumoniae systemic infection results in severe lung injury and significant bacterial accumulation in multiple organs, with the highest burden in the lungs. Deficiency of caspase-11 or NLRP3 led to prolonged survival, a reduction in pulmonary bacterial load, increased blood oxygen levels, and decreased IL-6 levels in the lungs compared to WT controls. Furthermore, caspase-11- and NLRP3-deficient macrophages exhibited elevated mitochondrial ROS production in response to K. pneumoniae, which correlated with more effective bacterial clearance.DiscussionThese results suggest that caspase-11 and NLRP3 contribute to K. pneumoniae-induced sepsis by impairing mitochondrial function and reducing ROS production in macrophages, thereby compromising bacterial clearance. The observed reduction in lung injury and increased survival in caspase-11- and NLRP3-deficient mice indicate that targeting these pathways may offer potential therapeutic strategies to improve host defense against systemic K. pneumoniae infection.

The NAIP/NLRC4 inflammasome plays a pivotal role in the defense against bacterial infections, with its in vivo physiological function primarily recognized as driving inflammation in immune cells. Acute lung injury (ALI) is a leading cause of mortality in sepsis. In this study, we identify that the NAIP/NLRC4 inflammasome is highly expressed in both macrophages and pulmonary fibroblasts and that pyroptosis of these cells plays a critical role in lung injury. Mice challenged with gram-negative bacteria or flagellin developed lethal lung injury, characterized by reduced blood oxygen saturation, disrupted lung barrier function, and escalated inflammation. Flagellin-induced lung injury was protected in caspase-1 or GSDMD-deficient mice. These findings enhance our understanding of the NAIP/NLRC4 inflammasome's (patho)physiological function and highlight the significant role of inflammasome activation and pyroptosis in ALI during sepsis.

Background:Irisin, a novel myokine, has garnered significant attention for its roles in metabolic regulation and anti-inflammatory responses. Sepsis disrupts the intestinal microenvironment, exacerbating its progression and highlighting the need for novel therapeutic approaches. This study aims to investigate whether irisin exerts protective effects against lipopolysaccharide (LPS)-induced intestinal injury in septic conditions and to explore the underlying mechanisms involving the gut microbiota. Methods:To induce sepsis, C57BL/6 mice were injected intraperitoneally with LPS at a dose of 10 mg/kg, and then administered with 1 µg/kg of irisin. The Activity levels and 7-day survival rate were recorded. The intestinal expression of irisin/FNDC5 was assessed using Western blotting and immunofluorescence staining. Inflammatory factors were measured using enzyme-linked immunosorbent assay (ELISA). Peripheral blood bacteria were cultured on blood agar plates. Intestinal histomorphology was analyzed via hematoxylin and eosin (H&E) staining. The expression of occludin and apoptotic-related proteins was determined by Western blot, and apoptotic cells were detected using the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) method. The intestinal microbiota was analyzed through 16S rRNA amplicon sequencing. Results:Irisin improved the survival state and rate of LPS-induced septic mice. It restored endogenous irisin/FNDC5 levels in intestinal tissues, mitigated intestinal barrier injury, and alleviated bacteremia following sepsis treatment. Furthermore, irisin exhibited anti-inflammatory properties by increasing the levels of IL-22 while decreasing those of TNF-α and IL-6, as well as anti-apoptotic effects by increasing levels of pro-caspase-3 and Bcl-2 while decreasing cleaved caspase-3, Bax, and the positive density of apoptotic cells. Additionally, it regulated intestinal microbiota dysfunction. Conclusion:Irisin effectively treats septic acute intestinal injury by reducing apoptosis and inflammation, with the intestinal microbiota likely playing a crucial role. This finding offers a novel approach to clinical management of sepsis.

The field of non-coding RNA research is advancing at a breathtaking pace, continually uncovering new layers of regulatory complexity and functional diversity [...]

Toll-like receptor 3 (TLR3) initiates antiviral and inflammatory responses exclusively through the adaptor protein TRIF (TIR-domain-containing adapter-inducing interferon-β). In contrast, MyD88 (myeloid differentiation primary response 88), a central adaptor for most other TLRs, is traditionally considered dispensable for TLR3 signaling. Here, we demonstrate that MyD88 directly contributes to TLR3-mediated NF-κB activation and cytokine production in macrophages. Bone marrow-derived macrophages (BMDMs) from MyD88 deficient mice exhibited significantly attenuated NF-κB activation in response to the TLR3 agonist polyinosinic–polycytidylic acid (poly(I:C)) compared to wild-type cells, as evidenced by the reduced phosphorylation of NF-κB p65 and IκBα, as well as IκBα degradation. Consistently, pro-inflammatory cytokine production, including IL-6, TNF-α, and IFN-β, was attenuated in MyD88-deficient BMDMs in vitro following stimulation by poly(I:C) or poly(A:U), another TLR3 agonist. Blood concentrations of IL-6, TNF-α, and IFN-β were significantly reduced in both TRIF-deficient mice and MyD88-deficient mice challenged by the i.p. injection of poly(I:C). Mechanistic analyses revealed that MyD88 physically associates with activated TLR3 upon poly(I:C) stimulation, and that TLR3 engagement triggered MyD88 oligomerization, which was absent in TLR3 or TRIF deficient macrophages. Our findings highlight a previously unrecognized dual-adaptor mechanism for TLR3, wherein MyD88 recruitment amplifies NF-κB signaling dynamics by bridging TLR3 to the canonical NF-κB activation cascade and robust cytokine induction. This study expands the paradigm of TLR3 signaling by establishing MyD88 as a direct contributor to TLR3-driven innate immune responses, offering new insight into cross-talk between MyD88-dependent and -independent pathways.

Mammalian spermatogenesis comprises three phases: the mitotic phase of spermatogonia (involving self-renewal and proliferation), the meiotic phase of spermatocytes (producing haploid round spermatids), and the spermiogenic phase (transforming round spermatids into spermatozoa). This process depends critically on maintaining a normal transcriptome and proteome. While recent studies demonstrated that conditional knockout of Exosc10 in male germ cells prior to meiosis disrupts meiosis, causing spermatogenic defects and male infertility, the role of EXOSC10 in spermatogonial maintenance remained unknown. This study reveals the critical role of EXOSC10 in maintaining mouse spermatogonia. Knockout of Exosc10 in embryonic (E15.5) male germ cells using Ddx4-Cre mice disrupts spermatogonial maintenance. This is manifested by reduced germ cell proliferation, arrested spermatogenesis, failed sperm production, and consequent male infertility. Transcriptomic and proteomic analyses confirmed that Exosc10 deficiency disrupts the expression of genes and proteins associated with spermatogenesis, ribosome biogenesis, germline stem cell maintenance, and regulation of reproductive processes, thereby impairing spermatogonial maintenance and blocking spermatogenesis. In summary, this study highlights that EXOSC10 safeguards normal sperm production and male fertility by maintaining the transcriptome and proteome essential for spermatogenesis, particularly at the spermatogonial stage.

In this study, we observed worsening metabolic crosstalk in mouse models with concomitant metabolic disorders such as hyperhomocysteinemia (HHcy), hyperlipidemia, and hyperglycemia and in human coronary artery disease by analyzing metabolic profiles. We found that HHcy worsening is most sensitive to other metabolic disorders. To identify metabolic genes and metabolites responsible for the worsening metabolic crosstalk, we examined mRNA levels of 324 metabolic genes in Hcy, glucose-related and lipid metabolic systems. We examined Hcy-metabolites (Hcy, SAH and SAM) by LS-ESI-MS/MS in 6 organs (heart, liver, brain, lung, spleen, and kidney) from C57BL/6J mice. Through linear regression analysis of Hcy-metabolites and metabolic gene mRNA levels, we discovered that SAH-responsive genes were responsible for most metabolic changes and all metabolic crosstalk mediated by Serine, Taurine, and G3P. SAH-responsive genes worsen glucose metabolism and cause upper glycolysis activation and lower glycolysis suppression, indicative of the accumulation of glucose/glycogen and G3P, Serine synthesis inhibition, and ATP depletion. Insufficient Serine due to negative correlation of PHGDH with SAH concentration may inhibit the folate cycle and transsulfurarion pathway and consequential reduced antioxidant power, including glutathione, taurine, NADPH, and NAD+. Additionally, we identified SAH-activated pathological TG loop as the consequence of increased fatty acid (FA) uptake, FA β-oxidation and Ac-CoA production along with lysosomal damage. We concluded that HHcy is most responsive to other metabolic changes in concomitant metabolic disorders and mediates worsening metabolic crosstalk mainly via SAH-responsive genes, that organ-specific Hcy metabolism determines organ-specific worsening metabolic reprogramming, and that SAH, acetyl-CoA, Serine and Taurine are critical metabolites mediating worsening metabolic crosstalk, redox disturbance, hypomethylation and hyperacetylation linking worsening metabolic reprogramming in metabolic syndrome.

BackgroundCalcific aortic valve disease (CAVD) is one of the most prevalent valvular diseases and is the second most common cause for cardiac surgery. However, the mechanism of CAVD remains unclear. This study aimed to investigate the role of pyroptosis-related genes in CAVD by performing comprehensive bioinformatics analysis.MethodsThree microarray datasets (GSE51472, GSE12644 and GSE83453) and one RNA sequencing dataset (GSE153555) were obtained from the Gene Expression Omnibus (GEO) database. Pyroptosis-related differentially expressed genes (DEGs) were identified between the calcified and the normal valve samples. LASSO regression and random forest (RF) machine learning analyses were performed to identify pyroptosis-related DEGs with diagnostic value. A diagnostic model was constructed with the diagnostic candidate pyroptosis-related DEGs. Receiver operating characteristic (ROC) curve analysis was performed to estimate the diagnostic performances of the diagnostic model and the individual diagnostic candidate genes in the training and validation cohorts. CIBERSORT analysis was performed to estimate the differences in the infiltration of the immune cell types. Pearson correlation analysis was used to investigate associations between the diagnostic biomarkers and the immune cell types. Immunohistochemistry was used to validate protein concentration.ResultsWe identified 805 DEGs, including 319 down-regulated genes and 486 up-regulated genes. These DEGs were mainly enriched in pathways related to the inflammatory responses. Subsequently, we identified 17 pyroptosis-related DEGs by comparing the 805 DEGs with the 223 pyroptosis-related genes. LASSO regression and RF algorithm analyses identified three CAVD diagnostic candidate genes (TREM1, TNFRSF11B, and PGF), which were significantly upregulated in the CAVD tissue samples. A diagnostic model was constructed with these 3 diagnostic candidate genes. The diagnostic model and the 3 diagnostic candidate genes showed good diagnostic performances with AUC values >0.75 in both the training and the validation cohorts based on the ROC curve analyses. CIBERSORT analyses demonstrated positive correlation between the proportion of M0 macrophages in the valve tissues and the expression levels of TREM1, TNFRSF11B, and PGF.ConclusionThree pyroptosis-related genes (TREM1, TNFRSF11B and PGF) were identified as diagnostic biomarkers for CAVD. These pyroptosis genes and the pro-inflammatory microenvironment in the calcified valve tissues are potential therapeutic targets for alleviating CAVD.