Diazepam (DZP), one of the most widely prescribed benzodiazepines (BZDs), is commonly used clinically to treat anxiety and epilepsy by reducing neuronal excitability. However, its potential ecological toxicity remains poorly understood. In this study, adult female zebrafish were exposed to environmentally relevant concentrations of DZP for 28 days, and hepatic responses were evaluated through integrated biochemical, transcriptional, and untargeted metabolomics analyses. DZP exposure altered antioxidant enzyme activities and increased oxidative stress markers, indicating disruption of hepatic redox homeostasis. Significant alterations were also observed in triglycerides (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and glucose (Glu). These physiological alternations were accompanied by modified expression of genes associated with glucose and lipid metabolism. Untargeted metabolomics analysis further revealed extensive metabolic reprogramming after DZP exposure. KEGG enrichment analysis indicated significant disturbances in energy and amino acid metabolism, with histidine metabolism identified the most prominently affected pathway. Alterations in metabolites associated with glutathione synthesis, membrane phospholipids, and purine metabolism further suggested widespread metabolic dysregulation in the liver following DZP exposure. Collectively, these findings demonstrate that environmentally relevant DZP exposure disrupts hepatic metabolism and induces oxidative stress in zebrafish, providing new insights into the ecotoxicological effects of DZP on aquatic organisms.

The widespread consumption and incomplete wastewater removal of the psychoactive drug diazepam (DZP) have led to its ubiquitous presence in aquatic environments, posing profound ecological risks. While the direct toxicity of DZP is well-documented, its chronic and mechanistic impacts, particularly those concerning the gut-brain axis and intergenerational effects on offspring, remain largely unexplored. To investigate the multigenerational toxicity of DZP, adult female zebrafish were chronically exposed to environmentally relevant concentrations (1 and 10 μg/L) for 28 days, and their offspring were subsequently collected. Our findings revealed that DZP exposure induced severe gut damage and disrupted the gut microbiota and metabolite profiles in female zebrafish. This microecological dysbiosis was linked to impaired oocyte development and reduced brain γ-aminobutyric acid (GABA) levels, culminating in reproductive dysfunction and abnormal neurobehavior. Crucially, these toxicological effects were maternally transferred to the offspring, which exhibited corresponding developmental abnormalities and neurobehavioral deficits. Overall, this study demonstrates that environmentally relevant concentrations of DZP exert covert but profound intergenerational toxicity, potentially mediated by disruptions in the gut-brain axis, making it highly necessary to establish stricter regulatory guidelines regarding the discharge of psychoactive drugs into water environments.

Purpose:Severe asthma (SA) is a heterogeneous disease with unmet therapeutic needs. Metabolic dysregulation involving glycolysis and cholesterol synthesis is implicated in its pathogenesis. This study aimed to systematically profile the glycolysis-cholesterol synthesis axis in SA to identify distinct metabolic subtypes and explore their potential therapeutic implications. Patients and Methods:We analyzed sputum mRNA expression from 93 SA patients and 16 healthy controls (HC) in the U-BIOPRED cohort (GSE76262). First, differentially expressed genes (DEGs) in the glycolysis-cholesterol synthesis axis were identified between SA patients and HC. Unsupervised consensus clustering was then applied to these DEGs within the SA cohort to define metabolic subtypes. Immune microenvironment features were characterized using single-sample gene set enrichment analysis. A random forest (RF) model was built to distinguish subtypes, and candidate therapeutic compounds were screened for each subtype via the Connectivity Map (CMap) database based on subtype-specific gene signatures. Results:This study revealed that, 47 genes in the glycolysis-cholesterol synthesis axis were dysregulated in SA when compared to HC. Clustering of SA based on these DEGs revealed two distinct metabolic subtypes (Cluster 1, n=52; Cluster 2, n=41). Cluster 1 exhibited an up-regulated axis with an immune landscape enriched for NK and gamma-delta T cells, linked to oxidative phosphorylation. Conversely, Cluster 2 displayed a down-regulated axis with enrichment of eosinophils, neutrophils, mast cells, and Th17 cells, associated with calcium and cAMP signaling. An RF model utilizing PPP2CB and SEH1L achieved accurate subtype discrimination (AUC=0.888), validated by decreased protein expression in an SA murine model. Exploratory drug screening suggested dipeptidyl peptidase inhibitor and mTOR inhibitor were identified as candidate compounds for Cluster 1, while leukotriene receptor antagonist and calcium channel blocker were suggested for Cluster 2 via CMap analysis. Conclusion:This study reveals two novel metabolic subtypes in SA, which offers a classification model and suggests potential targeted therapeutics for personalized management.

As an emerging alternative to perfluorooctanoic acid (PFOA), hexafluoropropylene oxide trimeric acid (HFPO-TA) has attracted increasing attention due to its widespread environmental occurrence and strong propensity for bioaccumulation. Nevertheless, the reproductive toxicity of PFOA and HFPO-TA in aquatic animals, as well as their effects on subsequent generations, remains poorly understood. To elucidate the reproductive effects of these compounds, adult male and female zebrafish were exposed to PFOA or HFPO-TA at concentrations of 0.5 and 50 µg/L, after which offspring were generated and collected for subsequent analyses. The findings demonstrated that both compounds disrupted mitochondrial integrity in gonadal cells, induced endocrine disturbances, altered sex hormone levels, and caused abnormal gonadal cell development, collectively impairing reproductive function. HFPO-TA elicited more severe endocrine disruption and gonadal damage in parental zebrafish than PFOA, with females exhibiting greater susceptibility to endocrine perturbation. In the offspring larvae, profound transcriptomic reprogramming was observed, concomitant with disturbances in growth and development, redox homeostasis, apoptotic signaling, and swimming performance. Notably, PFOA exerted stronger intergenerational toxicity, affecting offspring more profoundly than HFPO-TA. In summary, while HFPO-TA produced a more pronounced reproductive toxicity phenotype in parental zebrafish, PFOA posed a substantially greater risk to future generations.

Background: Tumor-associated macrophages (TAMs) are prevalent in advanced ovarian cancer tissues and ascites, significantly influencing disease prognosis. However, the mechanisms driving TAM polarization and their tumor-promoting effects remain poorly understood. Methods: The subcellular distribution of SNX10 in ovarian cancer tissues was analyzed using single-cell datasets (GSE147082, GSE58937). The Kaplan–Meier Plotter and GEPIA2 databases were used to evaluate SNX10’s prognostic relevance. Lentivirus-mediated SNX10 overexpression in THP-1 cells was employed in tumor cell–macrophage co-culture experiments. Transwell assays and flow cytometry assessed SNX10’s effects on ovarian cancer cell metastasis and cisplatin-induced apoptosis. RNA sequencing, Western blotting, lysosomal pH detection, lipid droplet staining, and RT-qPCR were performed to explore SNX10’s molecular mechanisms in TAM polarization and immune modulation. Results: SNX10 was specifically expressed in TAMs, promoting their polarization into the M2 phenotype. This enhanced the migration and invasion of ovarian cancer cell lines A2780 and A2780/CP70 while reducing cisplatin-induced apoptosis. SNX10 decreased lipid droplet content, downregulated p-mTOR1, and impaired lysosomal function in TAMs. Additionally, SNX10 differentially modulated PD-L1 mRNA expression in platinum-sensitive and platinum-resistant ovarian cancer cells. Conclusions: SNX10 regulates the mTOR1/lysosome pathway in TAMs, influencing lipid metabolism and indirectly modulating ovarian cancer cell metastasis. It also alters PD-L1 mRNA expression, suggesting a role in shaping the tumor immune microenvironment.

BACKGROUND:G protein-coupled receptor 37 (GPR37) has recently been earmarked as a rising candidate for use as a marker for central nervous system (CNS) demyelinating illness. In this study, we explored the ability of serum GPR37 to assess disease activity, functional impairment, and the efficacy of therapeutic interventions in patient cases of neuromyelitis optica spectrum disorder (NMOSD) and multiple sclerosis (MS). METHODS:We prospectively enrolled 144 cases of NMOSD and 132 cases of MS, as well as 160 controls (healthy individuals and cases of other neural disorders). Serum GPR37 levels were measured using ELISA. Longitudinal changes in GPR37 were evaluated in patients receiving acute-phase treatment (MS: n = 22; NMOSD: n = 28) and in those undergoing stable-phase follow-up after six months of maintenance therapy (MS: n = 18; NMOSD: n = 20). RESULTS:Compared to healthy controls, serum GPR37 was markedly decreased in MS and NMOSD. The reduction was most pronounced in NMOSD, distinguishing it from MS and other neurological diseases. Univariable and multivariable analyses revealed that, in patients with NMOSD, recent relapses were associated with decreased GPR37 levels. Additionally, GPR37 levels were negatively correlated with Expanded Disability Status Scale (EDSS) scores. Furthermore, serum GPR37 levels significantly increased after acute-phase treatment in both the MS and NMOSD groups, and this upward trend persisted over the subsequent six months of maintenance therapy. CONCLUSION:Serum GPR37 is a promising biomarker for the diagnosis of NMOSD and its differentiation from other neurological disorders. It also holds potential for assessing disease activity, disability severity, and treatment response in clinical practice, although the generalizability of these findings may be constrained by the single-center design.

Background:The anti-inflammatory effects of budesonide (BUN) and N-acetylcysteine (NAC) attenuate acute lung injury (ALI). The aim of this study was to investigate the effects of combination therapy consisting of BUN and NAC on ALI and the underlying mechanisms. Methods:In vitro and in vivo models of ALI were generated by LPS induction. Western blotting was used to detect the expression levels of pyroptosis-related proteins and inflammation-related factors, and RT-qPCR was used to detect the expression of miR-381. Cell proliferation and apoptosis were detected by CCK-8 and flow cytometry, respectively. ELISA was used to detect the levels of inflammation-related factors. HE staining was used to detect lung injury. Results:The results showed that LPS effectively induced pyroptosis in cells and promoted the expression of pyroptosis-related proteins (Caspase1, Gasdermin D and NLRP3) and inflammatory cytokines (TNF-α, IL-6 and IL-1β). The combination of BUN and NAC significantly alleviated LPS-induced pyroptosis and inflammation. In addition, the combination of BUN and NAC effectively promoted miR-381 expression. Transfection of miR-381 mimics effectively alleviated LPS-induced pyroptosis and inflammation, while transfection of miR-381 inhibitors had the opposite effect. miR-381 negatively regulates NLRP3 expression. Treatment with a miR-381 inhibitor or pc-NLRP3 reversed the effects of the combination of BUN and NAC. In a mouse model of ALI, the combination of BUN and NAC effectively improved lung injury, while treatment with a miR-381 inhibitor or pc-NLRP3 effectively reversed this effect. Conclusion:Overall, this study revealed that BUN + NAC inhibits the activation of NLRP3 by regulating miR-381, thereby alleviating ALI caused by pyroptosis-mediated inflammation.

Sodium hypochlorite (NaClO) and cadmium (Cd) are widely co-occurring in natural aquatic environment; however, no study has been conducted on effects of their combined exposure on aquatic organisms. To assess effects of exposure to NaClO and Cd in zebrafish larvae, we designed six treatment groups, as follows: control group, NaClO group (300 μg/L), 1/100 Cd group (48 μg/L), 1/30 Cd group (160 μg/L), NaClO+1/100 Cd group, and NaClO+1/30 Cd group analyzed behavior, neurological function and cardiac function. Results revealed that exposure to 1/30 Cd and NaClO+1/30 Cd caused abnormal embryonic development in larvae by altering body morphology and physiological indicators. Combined exposure to NaClO and 1/30 Cd affected the free-swimming activity and behavior of larvae in response to light-dark transition stimuli. Moreover, exposure to 1/30 Cd or NaClO+1/30 Cd resulted in a significant increase in tyrosine hydroxylase and acetylcholinesterase activities, as well as significant changes of various neurotransmitters. Lastly, exposure to 1/30 Cd or NaClO+1/30 Cd influenced the transcription of cardiac myosin-related genes and disturbed the myocardial contractile function. Altogether, our results suggested that combined exposure to NaClO and Cd induced oxidative damage in larvae, resulting in detrimental effects on nervous system and cardiac function, thus altering their swimming behavior.

Phytosterol organic acid esters are important food resources and the components of biomembrane structure. Due to the lack of extraction and synthesis techniques, more research has been focused on phytosterols, and the research on phytosterol acid esters have encountered a bottleneck, but phytosterol acid esters confer substantial benefits to human health. In this study, stigmasteryl vanillate (VAN), stigmasteryl protocatechuate (PRO) and stigmasteryl sinapate (SIN) were prepared through the Steglich reaction. The processes are promotable and the products reach up to 95% purity. In addition, their stability was evaluated by differential scanning calorimetry and thermogravimetric analysis. HPLC analysis revealed an enhancement in water solubility after esterification with phenolic acid. In an in vitro digestion model, the bioaccessibility of stigmasteryl phenolates was significantly higher than that of stigmasterols (STIs). Regarding the anti-inflammatory properties, VAN, PRO, and SIN exhibit superior effects against TNF-α induced pro-inflammatory responses compared to STI. All stigmasteryl phenolates supplementation increased the ATP production, the basal, and maximal oxygen consumption rate in mitochondrial stress test. Overall, we present a synthesis method for stigmasteryl phenolates. It will contribute to the development and research of phytosterol acid ester analysis, functions and utilization in food. Moreover, the nutrient-stigmasterol hybrids tactic we have constructed is practical and can become a targeted mitochondrial delivery strategy with enhanced anti-inflammatory effects.

Prothioconazole (PTC), as a popular triazole fungicide, with its main metabolite prothioconazole desthio (PTCd), have attracted widespread concern due to their widely use and toxicological effects on non-target organisms. However, toxic effects of study analyzed PTC and PTC-d on the hepatic metabolism of mammalian still remains unclear. In this study, we conducted the study of the C57BL/6 mice which oral exposure to 30 mg/kg PTC and PTC-d via metabolomic analysis. In the liver, the metabolomics profile unveiled that exposure to 30 mg/kg PTC and PTC-d led to significantly altered 13 and 28 metabolites respectively, with 6 metabolites in common including significant decreased D-Fructose, Glutathione, showing the change of carbohydrate, lipid and amino acid metabolism. Via the further exploration of genes related to hepatic glycolipid metabolism and the biomarkers of oxidative stress, we found that liver was potentially damaged after exposure to 5 and 30 mg/kg PTC and PTC-d. Particularly, it was proved that PTC-d caused more adverse effect than its parent compound PTC on hepatotoxicity, and high concentration PTC or PTC-d exposure is more harmful than low concentration exposure.

Epoxiconazole (EPX), a triazole fungicide, is widely used in agriculture to control pests and diseases. High residual and occupational exposure to EPX increases health risks, and evidence of potential harm to mammals remains to be added. In the present study, 6-week-old male mice were exposed to 10 and 50 mg/kg bw EPX for 28 days. The results showed that EPX significantly increased the liver weights. EPX also decreased the mucus secretion of the colon and altered intestinal barrier function in mice including a reduced expression of some genes (Muc2, meprinβ, tjp1). Moreover, EPX altered the composition and abundance of gut microbiota in the colon of mice. The alpha diversity indices (Shannon, Simpson) in the gut microbiota increased after exposure to EPX for 28 days. Interestingly, EPX increased the ratio of Firmicutes to Bacteroides and the abundance of other harmful bacteria including Helicobacter and Alistipes. Based on the untargeted metabolomic analysis, it was found that EPX altered the metabolic profiles of the liver in mice. KEGG analysis of differential metabolites revealed that EPX disrupted the pathway related to glycolipid metabolism, and the mRNA levels of related genes were also confirmed. In addition, the correlation analysis showed that the most altered harmful bacteria were associated with some significantly altered metabolites. The findings highlight that EPX exposure changed the micro-environment and lipid metabolism disturbance. These results also suggest that the potential toxicity of triazole fungicides to mammals cannot be ignored.

The NLRP3 (NOD−, LRR− and pyrin domain-containing protein 3) inflammasome plays a pivotal role in defending the host against infection as well as sterile inflammation. Activation of the NLRP3 inflammasome is critically regulated by a de-ubiquitination mechanism, but little is known about how ubiquitination restrains NLRP3 activity. Here, we showed that the membrane-bound E3 ubiquitin ligase gp78 mediated mixed ubiquitination of NLRP3, which inhibited NLRP3 inflammasome activation by suppressing the oligomerization and subcellular translocation of NLRP3. In addition, the endoplasmic reticulum membrane protein insulin-induced gene 1 (Insig-1) was required for this gp78–NLRP3 interaction and gp78-mediated NLRP3 ubiquitination. gp78 or Insig-1 deficiency in myeloid cells led to exacerbated NLRP3 inflammasome-dependent inflammation in vivo, including lipopolysaccharide-induced systemic inflammation and alum-induced peritonitis. Taken together, our study identifies gp78-mediated NLRP3 ubiquitination as a regulatory mechanism that restrains inflammasome activation and highlights NLRP3 ubiquitination as a potential therapeutic target for inflammatory diseases.

PurposeDiabetic heart failure (DHF) or cardiomyopathy is a common complication of diabetes; however, the underlying mechanism is not clear. In the present study, the authors searched for differentially expressed genes associated with DHF and the molecular types of immune cells based on bioinformatics.MethodsThe RNA expression dataset of DHF was obtained from the NCBI Gene Expression Omnibus (GEO) database. After preprocessing the data, the differentially expressed genes (DEGs) between the DHF group and the non-diabetic heart failure (NHF) group were screened and intersected with immune-related genes (IRGs) in the ImmPort database. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed using the DAVID tool. The ssGSEA algorithm was used to evaluate immune infiltration of the heart tissue in each group. In addition, the protein-protein interaction (PPI) network and miRNA-mRNA network were constructed using the STRING online website and Cytoscape program. Finally, validation analysis was performed using animal models.ResultsEight immune-related core genes were identified. GO and KEGG showed that core genes were mainly enriched in angiogenesis and cytokine-cytokine receptor interaction. Immune infiltration results showed that activated dendritic cells, central memory CD4 T cells, central memory CD8 T cells, myeloid-derived suppressor cells (MDSCs), neutrophils, and regulatory T cells may be involved in DHF. Neutrophils may play a key role in the pathogenesis of HF in diabetes.ConclusionImmune-related core genes and immune infiltrating cells provide a new perspective on the pathogenesis of DHF.

Immunometabolism contributes to inflammation, but how activated macrophages acquire extracellular nutrients to fuel inflammation is largely unknown. Here, we show that the plasma membrane potential (Vm) of macrophages mediated by Kir2.1, an inwardly-rectifying K+ channel, is an important determinant of nutrient acquisition and subsequent metabolic reprogramming promoting inflammation. In the absence of Kir2.1 activity, depolarized macrophage Vm lead to a caloric restriction state by limiting nutrient uptake and concomitant adaptations in nutrient conservation inducing autophagy, AMPK (Adenosine 5'-monophosphate-activated protein kinase), and GCN2 (General control nonderepressible 2), which subsequently depletes epigenetic substrates feeding histone methylation at loci of a cluster of metabolism-responsive inflammatory genes, thereby suppressing their transcription. Kir2.1-mediated Vm supports nutrient uptake by facilitating cell-surface retention of nutrient transporters such as 4F2hc and GLUT1 by its modulation of plasma membrane phospholipid dynamics. Pharmacological targeting of Kir2.1 alleviated inflammation triggered by LPS or bacterial infection in a sepsis model and sterile inflammation in human samples. These findings identify an ionic control of macrophage activation and advance our understanding of the immunomodulatory properties of Vm that links nutrient inputs to inflammatory diseases.

Goblet cells and their main secretory product, mucus, play crucial roles in orchestrating the colonic host-microbe interactions that help maintain gut homeostasis. However, the precise intracellular machinery underlying this goblet cell–induced mucus secretion remains poorly understood. Gasdermin D (GSDMD) is a recently identified pore-forming effector protein that causes pyroptosis, a lytic proinflammatory type of cell death occurring during various pathophysiological conditions. Here, we reveal an unexpected function of GSDMD in goblet cell mucin secretion and mucus layer formation. Specific deletion of Gsdmd in intestinal epithelial cells (Δ IEC ) led to abrogated mucus secretion with a concomitant loss of the mucus layer. This impaired colonic mucus layer in Gsdmd Δ IEC mice featured a disturbed host-microbial interface and inefficient clearance of enteric pathogens from the mucosal surface. Mechanistically, stimulation of goblet cells activates caspases to process GSDMD via reactive oxygen species production; in turn, this activated GSDMD drives mucin secretion through calcium ion–dependent scinderin-mediated cortical F-actin disassembly, which is a key step in granule exocytosis. This study links epithelial GSDMD to the secretory granule exocytotic pathway and highlights its physiological nonpyroptotic role in shaping mucosal homeostasis in the gut.

The NLRP3 inflammasome, a critical component of the innate immune system, induces caspase‐1 activation and interleukin‐1β maturation and drives cell fate toward pyroptosis. However, the mechanism of NLRP3 inflammasome activation still remains elusive. Here we provide evidence that AKT regulates NLRP3 inflammasome activation. Upon NLRP3 activation, AKT activity is inhibited by second stimulus‐induced reactive oxygen species. In contrast, AKT activation leads to NLRP3 inhibition and improved mitochondrial fitness. Mechanistically, AKT induces the phosphorylation of the DDX3X (DEAD‐box helicase 3, X‐linked), a recently identified NLRP3 inflammasome component, and impairs the interaction between DDX3X and NLRP3. Furthermore, an AKT agonist reduces NLRP3‐dependent inflammation in two in vivo models of LPS‐induced sepsis and Alum‐induced peritonitis. Altogether, our study highlights an important role of AKT in controlling NLRP3 inflammasome activation.

OBJECTIVEThe present study was performed to determine the clinical relevance of KLF7 in tongue squamous cell carcinoma (TSCC) and to characterize its potential function and mechanism of action.MATERIALS AND METHODSKLF7 expression was measured by RT-qPCR in 21 tongue cancer samples. The clinical relevance of KLF7 was analyzed in another cohort of 127 TSCC samples from a public database. Then, we performed RNA sequencing analysis in KLF7-overexpressing TSCC (SCC9 and CAL27) cells to define significantly altered pathways. The possible changes in migration and adhesion were then analyzed in KLF7-overexpressing and knockdown TSCC cells.RESULTSOur results showed that KLF7 mRNA expression was upregulated in TSCC and was significantly associated with the T and N stages. Patients with high KLF7 expression had worse overall survival. RNA sequencing and KEGG enriched pathway analysis showed that altered genes were enriched in extracellular matrix-receptor interactions and focal adhesions in both cell lines. KLF7-overexpressing TSCC cell lines showed enhanced migration capacity and cell adhesion ability, and knockdown of KLF7 expression decreased TSCC migration and adhesion ability.CONCLUSIONSWe concluded that KLF7 was overexpressed in TSCC and has prognostic value. KLF7 promoted TSCC migration and increased cell adhesion.

Metabolic and epigenetic reprogramming play pivotal roles in driving inflammation, but the precise regulatory mechanisms remain minimally understood. Here we show an ionic control mediated by macrophage Kir2.1, an inwardly-rectifying K+ channel, promting lipopolysaccharide (LPS)-induced inflammation. Kir2.1 blockade by the selective inhibitor ML133 or its specific deletion in macrophages suppressed the production of LPS-induced inflammatory factors, such as interleukin-1β, and protected mice from LPS-induced sepsis in vivo. Kir2.1 loss-of-function led to a nutrient starvation phenotype, with impaired glucose and serine-glycine-one-carbon metabolism, whose synergy promotes the generation of S-adenosylmethionine (SAM). Accordingly, reduced SAM availability by Kir2.1 blockade decreased histone methylation at key inflammatory effector loci, such as Il1b. Although the immunomodulatory effect of Kir2.1 was independent of modulation by Ca2+ flux and general signaling pathways, its loss-of-function led to a depolarized membrane potential (Vm) which decreased the surface expression of nutrient transporters, including GLUT1 and CD98. We thus identify an ionic control of metabolic- epigenetic reprogramming that links inflammation to Vm-mediated nutrient acquisition and identifies potential new strategies for anti-inflammatory therapy.

Immune cell function depends on specific metabolic programs dictated by mitochondria, including nutrient oxidation, macromolecule synthesis, and post-translational modifications. Mitochondrial adaptations have been linked to acute and chronic inflammation, but the metabolic cues and precise mechanisms remain unclear. Here we reveal that histone deacetylase 3 (HDAC3) is essential for shaping mitochondrial adaptations for IL-1β production in macrophages through non-histone deacetylation. In vivo, HDAC3 promoted lipopolysaccharide-induced acute inflammation and high-fat diet-induced chronic inflammation by enhancing NLRP3-dependent caspase-1 activation. HDAC3 configured the lipid profile in stimulated macrophages and restricted fatty acid oxidation (FAO) supported by exogenous fatty acids for mitochondria to acquire their adaptations and depolarization. Rather than affecting nuclear gene expression, HDAC3 translocated to mitochondria to deacetylate and inactivate an FAO enzyme, mitochondrial trifunctional enzyme subunit α. HDAC3 may serve as a controlling node that balances between acquiring mitochondrial adaptations and sustaining their fitness for IL-1β-dependent inflammation.

Double-positive (DP) thymocytes undergo positive selection to become mature single-positive CD4 + and CD8 + T cells in response to T cell receptor (TCR) signaling. Unlike mature T cells, DP cells must respond to low-affinity self-peptide-MHC ligands before full upregulation of their surface TCR expression can occur. Thus, DP thymocytes must be more sensitive to ligands than mature T cells. A number of molecules have been found that are able to enhance the strength of the TCR signal to facilitate positive selection. However, almost all of these molecules are also active in mature T cells. Themis (thymocyte expressed molecule involved in selection) and Tespa1 (thymocyte expressed positive selection associated 1) are two recently discovered molecules essential for optimal TCR signaling and thymocyte development. A deficiency in both molecules leads to defects in positive selection. Here, we compared the relative contributions of Themis and Tespa1 to positive selection in thymocytes. We show that Tespa1 deficiency led to more limited and specific gene expression profile changes in cells undergoing positive selection. In mixed bone marrow transfer experiments, Tespa1 −/− cells showed more severe defects in thymocyte development than Themis −/− cells. However, Tespa1 −/− cells showed a substantial degree of homeostatic expansion and became predominant in the peripheral lymphoid organs, suggesting that Tespa1 is a thymic-specific TCR signaling regulator. This hypothesis is further supported by our observations in Tespa1 conditional knockout mice, as Tespa1 deletion in peripheral T cells did not affect TCR signaling or cell proliferation. The different regulatory effects of Tespa1 and Themis are in accordance with their nonredundant roles in thymocyte selection, during which Tespa1 and Themis double knockouts showed additive defects.