Zhonghan Yang

Background: The resolution of inflammation is actively driven by omega‐3 polyunsaturated fatty acids (PUFAs) via their specialized pro‐resolving mediator (SPM) derivatives, including resolvin E1 (RvE1), whose role has been well established. However, clinical application of these mediators is hampered by inherent instability and elevated production costs. Methods: To surmount these obstacles, we have engineered a biosynthetic platform based on the probiotic Escherichia coli Nissle 1917 (EcN) that enables controlled, sustained RvE1 production through inducible expression of COX2 and 5‐LOX, designated EcN‐RvE1. The catalytic capacity, intestinal persistence, and therapeutic efficacy of the platform were evaluated in vitro and in LPS‐induced acute inflammation and DSS‐induced colitis murine models. Results: In this study, we validate the capacity of EcN‐RvE1 to catalyse the conversion of eicosapentaenoic acid (EPA) to RvE1 and confirm its ability to achieve long‐term intestinal persistence. In murine models of acute inflammation and colitis, EcN‐RvE1 exerts marked anti‐inflammatory and tissue‐protective effects, which are mediated by the regulation of inflammatory cytokine expression and the amelioration of gut microbiota dysbiosis. Moreover, EcN‐RvE1 using Euglena gracilis as a photosynthetic protist‐based source of PUFAs also exhibits protective anti‐inflammatory activity. Conclusion: Collectively, we report a probiotic engineering platform for the biosynthesis of RvE1, offering a novel strategy for harnessing the anti‐inflammatory potential of PUFAs derivatives in clinical settings.

Background: Growing evidence implicates enoyl-CoA hydratase domain-containing protein 2 (ECHDC2) in oncogenesis, yet its role in glioblastoma (GBM) remains undefined. We aimed to clarify the pathological significance and molecular mechanisms of ECHDC2 in GBM. Methods: Gene-expression profiles from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases were analyzed. Kaplan–Meier curves were used to evaluate the prognostic value of ECHDC2. Immune cell infiltration was quantified using CIBERSORT, single-sample gene-set enrichment analysis (ssGSEA), and ESTIMATE algorithms. Spearman’s correlation analysis was applied to assess the associations between ECHDC2 expression levels, immune checkpoint molecules, and immune cell subsets. To elucidate the functional relevance of ECHDC2, Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and gene-set enrichment analyses (GSEA) were performed, while protein-protein interaction (PPI) networks were investigated using the STRING database. Subsequently, ECHDC2 was knocked down or overexpressed in GBM cell lines, and its effects on cell proliferation and migration were determined using CCK-8, EdU, wound-healing, and Transwell migration assays. Results: Upregulated ECHDC2 expression was significantly correlated with unfavorable clinicopathological features and reduced overall survival (OS) in patients with GBM. High ECHDC2 expression was associated with increased infiltration of effector-memory CD8 ⁺ T cells (TEM) and plasmacytoid dendritic cells (pDCs). Enrichment analyses demonstrated that ECHDC2 is involved in tumor progression, with a particular focus on the PI3K/Akt signaling pathway. In vitro experiments showed that ECHDC2 knockdown suppressed the proliferation and migration of GBM cells. Conversely, ECHDC2 overexpression exerted the opposite effects on GBM cell proliferation and migration. Conclusion: ECHDC2 overexpression promotes GBM progression and portends poor prognosis. ECHDC2 may serve as both a prognostic biomarker and a therapeutic target in GBM.

Adipose tissue exhibits remarkable plasticity, defined as its capability of adapting to various environments and energy demands by undergoing shifts in phenotypic, metabolic characteristics, and structure. However, obesity often results in diminished adipose tissue plasticity. Long non-coding RNA (lncRNA) is a class of RNA defined as being more than 200 nucleotides long and lacking protein-coding capability. With advancements in ribosome profiling, mass spectrometry, and other research technologies, an increasing number of studies have confirmed that short open reading frames (less than 300 nucleotides) within lncRNAs have the capacity to encode functional micropeptides. In our study, we initially identified a biologically active micropeptide, lncRNA Esrp2-as-ORF1(LEAO), encoded by lncRNA Esrp2-as in mice. In diet-induced obesity (DIO)mice, this micropeptide demonstrates the ability to induce fat browning, enhance adipose tissue plasticity, and subsequently improve metabolic homeostasis. Additionally, we observed that the micropeptide LEAO exerts its effects on adipocytes by stimulating IL6 secretion in adipose tissue macrophages. In summary, LEAO may have therapeutic potential for obesity-related metabolic diseases.

Background: Obesity has emerged as a global health challenge. Although GLP‐1 receptor agonists are showing considerable promise in weight loss, their clinical utility is partly limited by gastrointestinal adverse reactions and non‐fat weight loss side effects. UCP1‐mediated adipose thermogenesis is a critical process for body temperature maintenance and weight management. However, the lack of effective and specific adipose thermogenesis therapies has restricted its clinical application. We aimed to explore the potential of inducing adipose‐specific UCP1 overexpression via modified plasmids as an innovative therapeutic approach for obesity. Methods: We replaced the cytomegalovirus (CMV) promoter in the plasmids with two types of adipose‐specific promoters: mouse adiponectin (mADP) promoter and human adiponectin (hADP) promoter, to selectively overexpress UCP1 in adipocytes. The expression level of UCP1, weight loss, metabolic homeostasis and adipose thermogenesis effects were evaluated by immunohistochemistry, western blot, weight measurements, thermography, and comprehensive lab animal monitoring system. Results: The experiments demonstrated that the mADP promoter‐modified plasmids failed to drive UCP1 overexpression. In contrast, the hADP promoter‐modified Ucp1 overexpression (hADP‐ Ucp1 OE) plasmids achieved robust adipose‐specific UCP1 protein expression both in vitro and in vivo. In vitro experiments revealed that delivery of the hADP promoter‐modified UCP1 overexpression (hADP‐ UCP1 OE) plasmids reduced lipid droplet size and enhanced energy consumption in human adipocytes. In obese mice, administration of the hADP‐ Ucp1 OE plasmids resulted in significant weight loss and improved metabolic homeostasis. Conclusions: These findings highlight the therapeutic potential of hADP‐ UCP1 OE plasmids in obesity management. Key points: The hADP promoter‐modified plasmids selectively overexpress protein in adipose tissue. Overexpression of UCP1 driven by hADP promoter induces thermogenesis in mouse and human adipocytes in vitro. The hADP‐ Ucp1 OE treatment promotes thermogenesis and energy expenditure in mice. The hADP‐ Ucp1 OE treatment restrains the development of obesity and glucose intolerance in mice.

The induction of adipose thermogenesis plays a critical role in maintaining body temperature and improving metabolic homeostasis to combat obesity. β3-adrenoceptor (β3-AR) is widely recognized as a canonical β-adrenergic G-protein-coupled receptor (GPCR) that plays a crucial role in mediating adipose thermogenesis in mice. Nonetheless, the limited expression of β3-AR in human adipocytes restricts its clinical application. The objective of this study was to identify a GPCR that is highly expressed in human adipocytes and to explore its potential involvement in adipose thermogenesis. Our research findings have demonstrated that the adhesion G-protein-coupled receptor A3 (ADGRA3), an orphan GPCR, plays a significant role in adipose thermogenesis through its constitutively active effects. ADGRA3 exhibited high expression levels in human adipocytes and mouse brown fat. Furthermore, the knockdown of Adgra3 resulted in an exacerbated obese phenotype and a reduction in the expression of thermogenic markers in mice. Conversely, Adgra3 overexpression activated the adipose thermogenic program and improved metabolic homeostasis in mice without exogenous ligand. We found that ADGRA3 facilitates the biogenesis of beige human or mouse adipocytes in vitro. Moreover, hesperetin was identified as a potential agonist of ADGRA3, capable of inducing adipocyte browning and ameliorating insulin resistance in mice. In conclusion, our study demonstrated that the overexpression of constitutively active ADGRA3 or the activation of ADGRA3 by hesperetin can induce adipocyte browning by Gs-PKA-CREB axis. These findings indicate that the utilization of hesperetin and the selective overexpression of ADGRA3 in adipose tissue could serve as promising therapeutic strategies in the fight against obesity.

The induction of adipose thermogenesis plays a critical role in maintaining body temperature and improving metabolic homeostasis to combat obesity. β3-adrenoceptor (β3-AR) is widely recognized as a canonical β-adrenergic G protein-coupled receptor (GPCR) that plays a crucial role in mediating adipose thermogenesis in mice. Nonetheless, the limited expression of β3-AR in human adipocytes restricts its clinical application. The objective of this study was to identify a GPCR that is highly expressed in human adipocytes and to explore its potential involvement in adipose thermogenesis. Our research findings have demonstrated that the adhesion G protein-coupled receptor A3 (ADGRA3), an orphan GPCR, plays a significant role in adipose thermogenesis through its constitutively active effects. ADGRA3 exhibited high expression levels in human adipocytes and mouse brown fat. Furthermore, the knockdown of Adgra3 resulted in an exacerbated obese phenotype and a reduction in the expression of thermogenic markers. Conversely, Adgra3 overexpression activated the adipose thermogenic program and improved metabolic homeostasis without an exogenous ligand supplementation. We found that ADGRA3 facilitates the biogenesis of beige adipocytes through the Gs-PKA-CREB axis. Moreover, hesperetin was identified as a potential agonist of ADGRA3, capable of inducing adipocyte browning and ameliorating insulin resistance. In conclusion, our study demonstrated that the overexpression of constitutively active ADGRA3 or the activation of ADGRA3 by hesperetin can induce adipocyte browning by Gs-PKA-CREB axis. These findings indicate that the utilization of hesperetin and the selectively overexpression of ADGRA3 in adipose tissue could serve as promising therapeutic strategies in the fight against obesity.

Background: Bladder cancer is a common tumor of the urinary system. Interleukin-25 is highly expressed in a variety of tumors and leads to poor prognosis. However, the expression and biological role of interleukin-25 in bladder cancer remain unclear. Methods: This study analyzed the differential expression of interleukin-25 (IL-25) and its receptor, interleukin-17 receptor B (IL-17RB), in tumor and adjacent tissues from bladder cancer patients by bioinformatics databases and immunohistochemistry. The impact of IL-25 on bladder cancer cell proliferation and migration was assessed in vitro. Moreover, we used wild-type and IL-25 knockout mice implanted with bladder cancer cells to investigate tumor growth in vivo. Gene set enrichment analysis (GSEA) was employed to elucidate the underlying mechanisms. We further explored the function of interleukin-25 in bladder cancer by cell viability assay and western blot. Angiogenesis was also compared between the wild-type and IL-25 knockout mice. Results: Immunohistochemistry indicated a significantly higher expression of IL-25 in bladder cancer tissues, which correlated with several prognostic markers. Analysis of The Cancer Genome Atlas (TCGA) database revealed that IL-17RB was highly expressed in bladder cancer, and may be related to overall survival of patients. In vitro, IL-25 enhanced the proliferation and migration of bladder cancer cells. IL-25 knockout markedly reduced tumor growth in a murine subcutaneous bladder cancer model. Mechanistic investigations revealed that IL-25 enhanced bladder cancer cell proliferation by activating the AKT (protein kinase B), mammalian target of rapamycin (mTOR) signaling pathway (AKT/mTOR signaling pathway). Additionally, angiogenesis was suppressed in IL-25 knockout mice bearing bladder tumors. Conclusion: Our findings suggest that IL-25 promotes bladder cancer cell proliferation by activating the AKT/mTOR signaling pathway and IL-25 knockout inhibits angiogenesis in bladder cancer. These findings suggest that IL-25 is a potential prognostic marker and therapeutic target in bladder cancer management.