The FASEB Journal最新文献

{"title":"Exploration of Potential Core Targets for Acute Liver Injury Based on a Novel Network Pharmacology Strategy Integrating the Common Efficacy and Mechanisms of Active Monomers.","authors":"Minling Lyu, Jing Li, Rui Hu, Mengqing Ma, Qi Huang, Kongli Fan, Yuhan Wang, Jiahua Yang, Benqiang Cai, Qibiao Wu, Xiaozhou Zhou","doi":"10.1096/fj.202504908R","DOIUrl":"10.1096/fj.202504908R","url":null,"abstract":"<p><p>Acute liver injury (ALI) is defined as rapidly progressing hepatic dysfunction or hepatocellular necrosis caused by drugs or chemicals, viral infection, or autoimmune diseases, among which drug-induced liver injury (DILI) is the major etiology. Numerous monomeric compounds have shown hepatoprotective effects in animal models; however, their therapeutic specificity is limited, and their clinical applicability remains restricted. This study moved beyond the single-compound paradigm and systematically identified key candidate hubs of ALI by integrating the shared efficacy and mechanisms of hepatoprotective monomers. Monomeric compounds with preclinically confirmed hepatoprotective effects were obtained from PubMed. Network pharmacology was used to identify overlapping targets between monomers and ALI. Transcriptomic datasets were analyzed to explore the differential expression of candidate targets. In vivo validation was conducted in C57BL/6 mice using APAP- and LPS/D-GalN-induced ALI models. Molecular docking and molecular dynamics (MD) simulations were conducted to predict compound-target interactions. A total of 186 active monomers and four hub genes (JUN, STAT3, ESR1, and CTNNB1) were identified. Across multiple GEO datasets, JUN was the only consistently upregulated gene. In vivo models confirmed robust activation of phosphorylated c-Jun. Docking and MD analysis indicated stable binding of Schisandrol A, Withaferin A, and Schizandrin to JUN. This integrated strategy revealed JUN as a key candidate molecular hub in ALI. This study not only provides new ideas for exploring the common mechanism of ALI but also offers clues for the development of JUN-targeted hepatoprotective agents.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"40 8","pages":"e71729"},"PeriodicalIF":4.2,"publicationDate":"2026-04-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147730494","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Oridonin-Loaded PDA@Gel@GO Nanocapsules Modulate NLRP3 and Epithelial Repair in Colitis.","authors":"Wenbo Yuan, Qirui Zhao, Jing Hua, Lingyao Xu, Yufan Wang, Tongtong Xu, Zuchao Du, Liping Wu, Daxiang Cui","doi":"10.1096/fj.202503622R","DOIUrl":"10.1096/fj.202503622R","url":null,"abstract":"<p><p>Ulcerative colitis (UC) is a chronic inflammatory bowel disease marked by sustained mucosal inflammation and epithelial barrier dysfunction, closely associated with aberrant NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome activation. Targeted oral therapies that simultaneously suppress inflammation and promote mucosal repair remain limited. We developed an orally administrable nanomicrocapsule system (PDA@Gel@GO) for colon-targeted delivery of oridonin, a natural diterpenoid with anti-inflammatory activity. The system was constructed by encapsulating galactosylated carboxymethyl chitosan-based oridonin nanomicelles (GO) within gelatin microspheres, followed by polydopamine coating to enhance gastrointestinal stability and mucosal adhesion. In a dextran sulfate sodium (DSS)-induced UC mouse model, PDA@Gel@GO exhibited preferential accumulation in inflamed colonic tissues and significantly improved body weight loss, disease activity index, and colon shortening. Mechanistic studies demonstrated that PDA@Gel@GO treatment suppressed NLRP3 inflammasome activation, as evidenced by reduced expression of NLRP3, ASC, caspase-1, and IL-1β, accompanied by attenuation of oxidative stress and restoration of intestinal barrier proteins. Transcriptomic analysis further revealed activation of gene programs associated with epithelial regeneration, cell cycle progression, and DNA repair, suggesting coordinated inflammation resolution and mucosal repair. Collectively, this study presents a macrophage-targeted, multilayered nanomicrocapsule platform that enhances the therapeutic efficacy of oridonin through precise colon targeting and integrated anti-inflammatory and tissue-protective effects, providing a promising strategy for precision treatment of UC.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"40 8","pages":"e71797"},"PeriodicalIF":4.2,"publicationDate":"2026-04-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13094459/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147730508","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Gentiopicroside Alleviates Type 2 Diabetes Mellitus by Ameliorating Hepatic Oxidative Stress via Activation of the PI3K/AKT/Nrf2 Signaling Pathway.","authors":"Xing Wang, Dongmei Long, Linlin Wu, Qitong Xie, Xingcan Peng, Siyao Luo, Huijuan Li, Maoting Zhou, Nan Guo","doi":"10.1096/fj.202600873R","DOIUrl":"https://doi.org/10.1096/fj.202600873R","url":null,"abstract":"<p><p>Gentiopicroside (GPS), derived from Gentiana manshurica, exhibits multiple pharmacological activities, such as anti-inflammatory and antioxidant effects, but its role and mechanisms in treating type 2 diabetes mellitus (T2DM) remain unclear. This study explored GPS's therapeutic effects and underlying mechanisms in vitro and in vivo. In a T2DM mouse model induced by a high-fat diet (HFD) and streptozotocin (STZ), the results demonstrated that GPS significantly relieved diabetic symptoms, corrected oral glucose tolerance impairment, enhanced insulin sensitivity, and ameliorated lipid metabolism disorders. GPS also increased the activities of antioxidant enzymes, elevated the levels of antioxidant substances, reduced malondialdehyde (MDA) in serum and liver, and ameliorated hepatic insulin resistance (IR). In palmitic acid (PA)-induced HepG2 cells, GPS dramatically inhibited PA-induced cytotoxicity and oxidative stress by increasing glutathione (GSH) levels and superoxide dismutase (SOD) activities while lowering malondialdehyde (MDA) levels, and reversed PA-induced IR. Mechanistically, GPS regulated protein expression in the PI3K/AKT/Nrf2 pathway and promoted Nrf2 nuclear translocation in both models. More importantly, further studies revealed that LY294002 (a PI3K inhibitor) or ML385 (an Nrf2 inhibitor) suppressed the protective effects of GPS against PA-induced hepatic oxidative damage. In conclusion, GPS alleviates hepatic oxidative damage, IR and delays the progression of T2DM through activating the PI3K/AKT/Nrf2 signaling pathway, providing an important foundation for further development and utilization of GPS. However, this study still has limitations including the lack of a positive control drug and insufficient exploration of the relevant molecular mechanisms, and will further improve the research and explore the clinical potential of GPS in the future.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"40 8","pages":"e71842"},"PeriodicalIF":4.2,"publicationDate":"2026-04-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147786805","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Lactate Activates TGF-β/SNAIL Signaling to Drive M2 Macrophage Polarization and CD8⁺ T Cell Exhaustion in Breast Cancer.","authors":"Peiling Zhu, Zhisheng Tian, Ying Wang, Haiying Liao, Zehui Mi","doi":"10.1096/fj.202502538RR","DOIUrl":"https://doi.org/10.1096/fj.202502538RR","url":null,"abstract":"<p><p>This study demonstrates that lactate promotes M2-like macrophage polarization by activating the TGF-β/SNAIL signaling axis, thereby weakening CD8⁺ T cell-mediated antitumor immunity and promoting breast cancer progression. In vitro experiments using bone marrow-derived macrophages (BMDMs) and THP-1 cells treated with 25 mM lactate revealed a marked increase in M2 markers (CD206, Arg-1, IL-10) and a reduction in M1 markers (iNOS, TNF-α, IL-12), confirmed by Western blotting and flow cytometry. RNA-Seq analysis identified TGF-β/SNAIL pathway activation, with increased TGFBR1/2 expression, Smad2/3 phosphorylation, and PI3K/AKT pathway enrichment. Functional studies revealed that lactate-polarized M2 macrophages impaired CD8⁺ T cell cytotoxicity (reduced IFN-γ, GzmB, PRF1; elevated PD-1, Tim-3) and disrupted mitochondrial metabolism. In vivo validation using a breast cancer xenograft model showed that lactate treatment increased tumor growth and angiogenesis (VEGF/CD31⁺), while TGF-β inhibition (SB431542) reversed these effects. Mechanistically, lactate-induced TGF-β/SNAIL signaling promoted EMT in cancer cells and created an immunosuppressive TME. These findings establish lactate as a critical metabolic regulator that coordinates macrophage polarization and T cell exhaustion through the TGF-β/SNAIL axis, highlighting this pathway as a promising therapeutic target for breast cancer immunotherapy.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"40 8","pages":"e71753"},"PeriodicalIF":4.2,"publicationDate":"2026-04-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147786782","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Distinct Effects of Maternal Stress and Exercise on Offspring Metabolic Health.","authors":"Felix T Yang, Joao V Esteves, Lisa A Baer, Kelsey M Pinckard, Branden G Verosky, Olivia A Togliatti, Caden Burkhart, Sara A Alzuhaili, Nickolai P Seculov, Tamar L Gur, Kristin I Stanford","doi":"10.1096/fj.202600159R","DOIUrl":"https://doi.org/10.1096/fj.202600159R","url":null,"abstract":"<p><p>Maternal physical activity enhances offspring metabolic resilience, whereas prenatal psychosocial stress predisposes offspring to lifelong metabolic dysfunction. Whether maternal exercise can attenuate the deleterious programming effects of stress has remained unclear. Here, we investigated the effect of maternal exercise and/or prenatal psychosocial stress on the metabolic health of male and female offspring. C57BL/6 dams were assigned to four conditions: sedentary/non-stressed (SED-NS), sedentary/stressed (SED-S), exercised/non-stressed (EX-NS), or exercised/stressed (EX-S). Male EX-NS offspring had improved glucose tolerance compared to SED-NS and SED-S; however these effects were blunted in male EX-S offspring, indicating that prenatal stress abolished this exercise-induced benefit. In contrast, there was no effect of maternal stress or exercise on glucose tolerance in female offspring. Maternal exercise chronically elevated basal corticosterone in male offspring, regardless of maternal stress. Maternal stress increased mineralocorticoid receptor (Nr3c2) mRNA and glucocorticoid receptor (NR3C1) protein expression in brown adipose tissue in male mice, while exercise restored NR3C1 expression. These findings indicate that prenatal stress blunts the metabolic benefits of maternal exercise in male offspring and is associated with altered corticosteroid signaling in brown adipose tissue. Together, the data identify a stress-exercise interaction that may influence offspring metabolic programming through tissue-specific regulation of glucocorticoid pathways.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"40 8","pages":"e71742"},"PeriodicalIF":4.2,"publicationDate":"2026-04-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13100692/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147788007","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Purification and Transcriptomic Characterization of Hypertrophied Hepatic Stellate Cells From CDAHFD Mouse Liver.","authors":"Marion Heckmann, Nour-El-Houda Djerir, Keola Greliche, Pierre-Henri Commere, Julien Fernandes, Guillaume Sarrabayrouse, Bernard Hainque, Pascal Bigey, Virginie Escriou, Céline Hoffmann","doi":"10.1096/fj.202502655RR","DOIUrl":"https://doi.org/10.1096/fj.202502655RR","url":null,"abstract":"<p><p>Hepatic stellate cells (HSC) are known for their major role in hepatic fibrosis. It is well established that upon liver injury, they undergo a transition from a quiescent state to an activated state and transdifferentiate into proliferative, fibrogenic myofibroblasts. Recently, it has been shown that different subpopulations of HSC co-exist during fibrogenesis and play different roles in the establishment of fibrosis. We previously highlighted, in murine model and human biopsies, a specific subpopulation of hypertrophied HSC (hypHSC) which exhibits exacerbated retinoid droplets and were closely associated with collagen fibers. The present study focuses on the molecular characterization of hypHSCs isolated from a murine model of metabolic liver fibrosis thanks to an experimental strategy we developed and described here. Liver dissociation followed by density gradient and fluorescence assisted cell sorting allowed us to obtain highly pure hypHSC preparations. Then, a transcriptomic analysis (bulk RNAseq) of hypHSCs versus quiescent HSCs purified from healthy mouse liver was performed. This showed that hypHSCs' molecular signature differs from HSC subtypes already described in the literature, with a \"hybrid\" profile involved in the regulation of the immune system, the remodeling of extracellular matrix and exhibiting a deregulation of lipid metabolism. Our study highlights that a phenotype-to-molecular approach can provide complementary elements to single-cell molecular approaches and provides additional insights into the plasticity of hepatic stellate cells in a context of hepatic fibrosis.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"40 8","pages":"e71754"},"PeriodicalIF":4.2,"publicationDate":"2026-04-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13104797/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147787600","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"CSPG4 Mediates Inflammatory, Cell Death, and Senescence Responses in Enteric Glia Exposed to Clostridioides difficile Toxins.","authors":"Francisco M S Marques, Deiziane V S Costa, Conceição S M Rebouças, Gerly A C Brito, Cirle A Warren, Renata F C Leitão","doi":"10.1096/fj.202600333R","DOIUrl":"10.1096/fj.202600333R","url":null,"abstract":"<p><p>Clostridioides difficile infection (CDI) is a persistent public health problem worldwide. C. difficile toxins A (TcdA) and B (TcdB) can affect enteric glial cells, an important component of the enteric nervous system. Here, we aimed to identify the profile of C. difficile toxin receptors expressed by enteric glia and investigate the contribution of the chondroitin sulfate proteoglycan receptors (CSPG4) in mediating inflammatory response, cell death, and senescence in enteric glia exposed to TcdA or TcdB. We found that human, mouse, and rat enteric glia express all of the previously recognized C. difficile toxin receptors. However, in vitro, C. difficile toxins significantly increased CSPG4 expression in enteric glia. In vivo, CDI also increased CSPG4 expression in colonic and cecal tissues. Blockage of CSPG4 significantly decreased IL-6 and S100B gene expression, as well as reduced the nuclear translocation of NFκB and STAT3 in enteric glia exposed to TcdB, but not TcdA. Interestingly, blockade of CSPG4 also decreased senescence-associated beta-galactosidase (SA-β-galactosidase) activity and other senescence markers (γH2AX, p16 and p21) in enteric glia exposed to TcdA or TcdB. However, CSPG4 blockade reduced cell death promoted by TcdA, but not TcdB. Taken together, these findings show that enteric glia, by expressing all known C. difficile toxin receptors, can respond directly to TcdA or TcdB. However, CSPG4 seems to play an important role in cell death and inflammatory response promoted by TcdA and TcdB, respectively, as well as senescence induced by both toxins.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"40 8","pages":"e71806"},"PeriodicalIF":4.2,"publicationDate":"2026-04-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13091310/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147724496","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The PPARβ/Delta-Induced Mesenchymal Stromal Cell Secretome Has Cytoprotective Effects via ANGPTL4 in a Pre-Clinical Model of Acute Lung Inflammation.","authors":"Courteney Tunstead, Molly Dunlop, Sinéad Ryan, Evelina Volkova, Evangeline Johnston, Sabrina Batah, Claudia C Dos Santos, Bairbre McNicholas, Claire Masterson, John G Laffey, Karen English","doi":"10.1096/fj.202504659R","DOIUrl":"10.1096/fj.202504659R","url":null,"abstract":"<p><p>Human bone marrow-derived mesenchymal stromal cells (hBM-MSCs) are known to exert immunomodulatory and pro-reparative effects in vivo. This makes hBM-MSCs an enticing therapeutic candidate for inflammatory diseases, such as acute respiratory distress syndrome (ARDS). The ARDS microenvironment is complex and contains an abundance of free fatty acids (FFAs), which are known to differentially impact MSC functionality. PPARβ/δ is a ubiquitously expressed nuclear receptor that is activated in response to FFA-binding. PPARβ/δ has been shown to impact the therapeutic efficacy of mouse MSCs. This study sought to investigate the impact of PPARβ/δ-modulation on human MSC functionality in vitro and in vivo. hBM-MSCs were exposed to a synthetic PPARβ/δ agonist/antagonist in the presence or absence of ARDS patient serum and the immunomodulatory and pro-reparative capacity of the MSC secretome was investigated using in vitro assays and a pre-clinical model of LPS-induced acute lung inflammation (ALI). Our results highlighted enhanced pro-reparative capacity of PPARβ/δ-agonized hBM-MSCs secretome in CALU-3 lung epithelial cells, mediated by MSC derived angiopoietin-like 4 (ANGPTL4). PPARβ/δ-induced ANGPTL4-high MSC secretome facilitated enhanced endothelial barrier integrity in the lungs of ALI mice. Therapeutic effects of PPARβ/δ-agonized hBM-MSCs secretome were further enhanced by licensing MSCs with human ARDS patient serum. ARDS-licensed PPARβ/δ-induced ANGPTL4-high MSC secretome had reduced clinical score and weight loss. The role ANGPL4 in these protective effects was confirmed using an anti-ANGPTL4 antibody. These findings conclude that the MSC secretome therapeutic effects can be enhanced both in vitro and in vivo through licensing strategies that upregulate the angiogenic factor ANGPTL4.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"40 8","pages":"e71820"},"PeriodicalIF":4.2,"publicationDate":"2026-04-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13094458/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147724558","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Lipocalin 2 Links Aquaporin 2⁺ Progenitor Cell-Derived Distal Renal Segments to Kidney Fibrosis.","authors":"Akaki Tsilosani, Chao Gao, Evelyn Mulroy, Noor Pirzada, Sana Shehzad, Shreya Das, Wei Sun, Enuo Chen, Sunny Guo, Nicole Knowles, Fizza Sajjad, Wenzheng Zhang","doi":"10.1096/fj.202600007RR","DOIUrl":"10.1096/fj.202600007RR","url":null,"abstract":"<p><p>Loss of Dot1l in Aquaporin 2⁺ progenitor cell (AP)-derived distal renal segments in Dot1l^ffAqp2cre (Dot1l^AC) promotes kidney fibrosis by upregulating endothelin 1 (ET1) through histone deacetylase 2 (HDAC2), defining a Dot1l-HDAC2-ET1 profibrotic pathway. Additionally, Lcn2 was also upregulated in Dot1l^AC mice. Here, we investigated whether and how Lipocalin 2 (Lcn2) contributes to this pathway. We performed in vivo studies using multiple mouse models with targeted deletion of Dot1l, Lcn2, or Edn1 in distal nephron segments or globally. Human CKD RNA-seq data analysis and in vitro studies in IMCD3 cells were conducted to validate the in vivo results. Because Lcn2^-/- mice are available and Lcn2 is barely detectable in IMCD3 cells, Lcn2 down- and upregulation experiments were performed in mice and IMCD3 cells, respectively. Several approaches, including immunofluorescence staining and in situ hybridization, were employed to identify and confirm the Lcn2 profibrotic role. Both Dot1l deletion in AP in Dot1l^AC mice and Dot1l silencing in IMCD3 cells upregulated Lcn2. Global deletion of Lcn2 in Dot1l^AC mice attenuated HDAC2 and ET1 levels and reduced kidney fibrosis, establishing Lcn2 as a downstream effector of Dot1l-deletion-induced kidney fibrosis. Although conditional knockout of Edn1 in Dot1l^AC mice showed Lcn2 expression comparable to that in Dot1l^AC mice, they exhibited reduced kidney fibrosis, suggesting that Lcn2 lies upstream of ET1-mediated injury. Reanalysis of a human chronic kidney disease (CKD) dataset revealed that downregulation of DOT1L was coupled with upregulation of LCN2, HDAC2, and ET1, recapitulating our findings in Dot1l^AC mice. These results were further validated in IMCD3 cells, in which Lcn2 was upregulated by Dot1l silencing, and addition of 293 T cells-secreted or recombinant Lcn2 led to increased HDAC2 and ET1 expression. Loss of Dot1l in AP upregulates Lcn2, which promotes kidney fibrosis by increasing HDAC2 and ET1. Our findings add Lcn2 as a new component to define a more complicated Dot1l-Lcn2-HDAC2-ET1 pro-fibrotic pathway that links AP-derived distal renal segments to kidney fibrosis.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"40 8","pages":"e71758"},"PeriodicalIF":4.2,"publicationDate":"2026-04-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147724655","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"p21 Knockdown as a Therapeutic Strategy for Focal Cartilage Injury Repair.","authors":"Leila Larijani, Aria Ahadzadeh Ardebili, Nini Ortiz Vales, Andreas T Stürmer, Roman J Krawetz","doi":"10.1096/fj.202600387RRR","DOIUrl":"10.1096/fj.202600387RRR","url":null,"abstract":"<p><p>Focal articular cartilage defects lack intrinsic regenerative capacity and can progress to osteoarthritis as no effective treatment exists to slow, stop or reverse cartilage degeneration. Cellular senescence, characterized by elevated p21 (CDKN1A) expression, impairs chondrocyte proliferation and cartilage repair. We hypothesized that targeted p21 suppression via lentiviral delivery would enhance cartilage regeneration. Lentiviral shRNA vectors targeting p21 or non-sense controls with a tdTomato reporter were constructed. In vitro, murine synovial progenitor cells and MC3T3E1 preosteoblasts were transduced and p21 mRNA was quantified by qRT-PCR. Cell cycle analysis was performed using flow cytometry. In vivo, immunocompromised (B6.Cg-Prkdc^scid/SzJ) and immunocompetent (C57BL6) mice received full-thickness cartilage defects followed by intraarticular p21 shRNA or nonsense control injection. Cartilage repair was assessed by Safranin O staining and tissue cytometry quantified lentiviral transduction and p21 knockdown in chondrocytes. In vitro, p21 shRNA achieved ~80% reduction in p21 mRNA and increased G2M phase cells. In vivo, p21 shRNA treatment significantly improved cartilage repair in both strains. Tissue cytometry revealed 90% transduction efficiency with p21⁺ cells reduced from ~90% to ~30%. Spearman correlation showed significant negative correlation between p21 expression and repair outcomes. Hepatic off-target transduction was minimal in uninjured animals but increased in injured animals, yet morphological hepatotoxicity was not observed. p21 knockdown via lentiviral shRNA effectively promotes cartilage regeneration, supporting the concept of targeting p21 and/or the p21 pathway as a therapeutic strategy for cartilage repair.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"40 8","pages":"e71814"},"PeriodicalIF":4.2,"publicationDate":"2026-04-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13094461/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147730478","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}