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In clinical settings, addressing large bone defects remains a significant challenge for orthopedic surgeons. The use of genetically modified bone marrow mesenchymal stem cells (BMSCs) has emerged as a highly promising approach for these treatments. Signal peptide‐CUB‐EGF domain‐containing protein 3 (SCUBE3) is a multifunctional secreted glycoprotein, the role of which remains unclear in human hBMSCs. This study used various experimental methods to elucidate the potential mechanism by which SCUBE3 influences osteogenic differentiation of hBMSCs in vitro. Additionally, the therapeutic efficacy of SCUBE3, in conjunction with porous GeLMA microspheres, was evaluated in vivo using a mouse bone defect model. Our findings indicate that SCUBE3 levels increase significantly during early osteogenic differentiation of hBMSCs, and that reducing SCUBE3 levels can hinder this differentiation. Overexpressing SCUBE3 elevated osteogenesis gene and protein levels and enhanced calcium deposition. Furthermore, treatment with recombinant human SCUBE3 (rhSCUBE3) protein boosted BMP2 and TGF‐β expression, activated mitophagy in hBMSCs, ameliorated oxidative stress, and restored osteogenic function through SMAD phosphorylation. In vivo, GELMA/OE treatment effectively accelerated bone healing in mice. In conclusion, SCUBE3 fosters osteogenic differentiation and mitophagy in hBMSCs by activating the BMP2/TGF‐β signaling pathway. When combined with engineered hydrogel cell therapy, it could offer valuable guidance for the clinical management of extensive bone defects. [ABSTRACT FROM AUTHOR]

Background: Aberrant TGF‐β signaling pathway can lead to invasive phenotype of colorectal cancer (CRC), resulting in poor prognosis. It is pivotal to develop an effective prognostic factor on the basis of TGF‐β‐related genes to accurately identify risk of CRC patients. Methods: We performed differential analysis of TGF‐β‐related genes in CRC patients from databases and previous literature to obtain TGF‐β‐related differentially expressed genes (TRDEGs). LASSO‐Cox regression was utilized to build a CRC prognostic feature model based on TRDEGs. The model was validated using two GEO validation sets. Wilcoxon rank‐sum test was utilized to test correlation of model with clinical factors. ESTIMATE algorithm and ssGSEA and tumor mutation burden (TMB) analysis were used to analyze immune landscape and mutation burden of high‐risk (HR) and low‐risk (LR) groups. CellMiner database was utilized to identify therapeutic drugs with high sensitivity to the feature genes. Results: We established a six‐gene risk prognostic model with good predictive accuracy, which independently predicted CRC patients' prognoses. The HR group was more likely to experience immunotherapy benefits due to higher immune infiltration and TMB. The feature gene TGFB2 could inhibit the efficacy of drugs such as XAV‐939, Staurosporine, and Dasatinib, but promote the efficacy of drugs such as CUDC‐305 and by‐product of CUDC‐305. Similarly, RBL1 could inhibit the drug action of Fluphenazine and Imiquimod but promote that of Irofulven. Conclusion: A CRC risk prognostic signature was developed on basis of TGF‐β‐related genes, which provides a reference for risk and further therapeutic selection of CRC patients. [ABSTRACT FROM AUTHOR]

Transforming growth factor‐β (TGF‐β) is a pleiotropic cytokine that modulates a wide variety of cellular responses by regulating target gene expression. It principally transmits signals via receptor‐activated transcription factors Smad2 and Smad3, which form trimeric complexes with Smad4 upon activation and regulate gene expression by binding to genomic DNA. Here, we examined the mechanisms by which TGF‐β regulates the transcription of target genes in a cell context‐dependent manner by screening a double‐stranded DNA oligonucleotide library for DNA sequences bound to endogenous activated Smad complexes. Screening was performed by cyclic amplification of selected targets (CASTing) using an anti‐Smad2/3 antibody and nuclear extracts isolated from three cell lines (A549, HepG2, and HaCaT) stimulated with TGF‐β. The preference of the activated Smad complexes for conventional Smad‐binding motifs such as Smad‐binding element (SBE) and CAGA motifs was different in HepG2 than in the other two cell lines, which may indicate the distinct composition of the activated Smad complexes. Several transcription factor‐binding motifs other than SBE or CAGA, including the Fos/Jun‐binding motifs, were detected in the enriched sequences. Reporter assays using sequences containing these transcription factor‐binding motifs together with Smad‐binding motifs indicated that some of the motifs may be involved in cell type‐dependent transcriptional activation by TGF‐β. The results suggest that the CASTing method is useful for elucidating the molecular basis of context‐dependent Smad signaling. [ABSTRACT FROM AUTHOR]

Hepatic stellate cell (HSC) activation is the essential pathological process of liver fibrosis (LF). The molecular mechanisms regulating HSC activation and LF are incompletely understood. Here, we explored the effect of transcription factor SRY‐related high mobility group box 7 (SOX7) on HSC activation and LF, and the underlying molecular mechanism. We found the expression levels of SOX7 were decreased in human and mouse fibrotic livers, particularly at the fibrotic foci. SOX7 was also downregulated in primary activated HSCs and TGF‐β1 stimulated LX‐2 cells. SOX7 knockdown promoted activation and proliferation of LX‐2 cells while inhibiting their apoptosis. On the other hand, overexpression of SOX7 suppressed the activation and proliferation of HSCs. Mechanistically, SOX7 attenuates HSC activation and LF by decreasing the expression of β‐catenin and phosphorylation of Smad2 and Smad3 induced by TGF‐β1. Furthermore, overexpression of SOX7 using AAV8‐SOX7 mouse models ameliorated the extent of LF in response to CCl4 treatment in vivo. Collectively, SOX7 suppressed HSC activation and LF. Targeting SOX7, therefore, could be a potential novel strategy to protect against LF. [ABSTRACT FROM AUTHOR]

The mammalian neocortex is formed by sequential radial migration of newborn excitatory neurons. Migrating neurons undergo a multipolar-to-bipolar transition at the subplate (SP) layer, where extracellular matrix (ECM) components are abundantly expressed. Here, we investigate the role of the ECM at the SP layer. We show that TGF-β signaling-related ECM proteins, and their downstream effector, p-smad2/3, are selectively expressed in the SP layer. We also find that migrating neurons express a disintegrin and metalloproteinase with thrombospondin motif 2 (ADAMTS2), an ECM metalloproteinase, just below the SP layer. Knockdown and knockout of Adamts2 suppresses the multipolar-to-bipolar transition of migrating neurons and disturbs radial migration. Time-lapse luminescence imaging of TGF-β signaling indicates that ADAMTS2 activates this signaling pathway in migrating neurons during the multipolar-to-bipolar transition at the SP layer. Overexpression of TGF-β2 in migrating neurons partially rescues migration defects in ADAMTS2 knockout mice. Our data suggest that ADAMTS2 secreted by the migrating multipolar neurons activates TGF-β signaling by ECM remodeling of the SP layer, which might drive the multipolar to bipolar transition. Synopsis: ADAMTS2 is secreted by migratory neurons during the formation of the mammalian neocortex. ADAMTS2 promotes neuronal migration by activating TGF-β signaling in migrating neurons during the multipolar-to-bipolar transition at the SP layer. ECM proteins are abundant in the subplate layer of the neocortex. Admts2 is upregulated during the migration of newborn neurons in the developing cortex. ADAMTS2 activates TGF-β signaling perhaps by ECM remodeling of the SP layer, which might facilitate the multipolar to bipolar transition. ADAMTS2 is secreted by migratory neurons during the formation of the mammalian neocortex. ADAMTS2 promotes neuronal migration by activating TGF-β signaling in migrating neurons during the multipolar-to-bipolar transition at the SP layer. [ABSTRACT FROM AUTHOR]

Several members of the transforming growth factor beta (TGF‐β) superfamily regulate the proliferation, differentiation, and function of bone‐forming osteoblasts and bone‐resorbing osteoclasts. However, it is still unknown whether Nodal, a member of the TGF‐β superfamily, serves a function in bone cells. In this study, we found that Nodal did not have any function in osteoblasts but instead negatively regulated osteoclast differentiation. Nodal inhibited RANKL‐induced osteoclast differentiation by downregulating the expression of pro‐osteoclastogenic genes, including c‐fos, Nfatc1, and Blimp1, and upregulating the expression of antiosteoclastogenic genes, including Bcl6 and Irf8. Nodal activated STAT1 in osteoclast precursor cells, and STAT1 downregulation significantly reduced the inhibitory effect of Nodal on osteoclast differentiation. These findings indicate that Nodal activates STAT1 to downregulate or upregulate the expression of pro‐osteoclastogenic or antiosteoclastogenic genes, respectively, leading to the inhibition of osteoclast differentiation. Moreover, the inhibitory effect of Nodal on osteoclast differentiation contributed to the reduction of RANKL‐induced bone loss in vivo. [ABSTRACT FROM AUTHOR]

Cadmium (Cd) is one of the most polluting heavy metal in the environment. Cd exposure has been elucidated to cause dysfunction of the glomerular filtration barrier (GFB). However, the underlying mechanism remains unclear. C57BL/6J male mice were administered with 2.28 mg/kg cadmium chloride (CdCl2) dissolved in distilled water by oral gavage for 14 days. The expression of SDC4 in the kidney tissues was detected. Human renal glomerular endothelial cells (HRGECs) were exposed to varying concentrations of CdCl2 for 24 h. The mRNA levels of SDC4, along with matrix metalloproteinase (MMP)‐2 and 9, were analyzed by quantitative PCR. Additionally, the protein expression levels of SDC4, MMP‐2/9, and both total and phosphorylated forms of Smad2/3 (P‐Smad2/3) were detected by western blot. The extravasation rate of fluorescein isothiocyanate–dextran through the Transwell was used to evaluate the permeability of HRGECs. SB431542 was used as an inhibitor of transforming growth factor (TGF)‐β signaling pathway to further investigate the role of TGF‐β. Cd reduced SDC4 expression in both mouse kidney tissues and HRGECs. In addition, Cd exposure increased permeability and upregulated P‐Smad2/3 levels in HRGECs. SB431542 treatment inhibited the phosphorylation of Smad2/3, Cd‐induced SDC4 downregulation, and hyperpermeability. MMP‐2/9 levels increased by Cd exposure was also blocked by SB431542, demonstrating the involvement of TGF‐β/Smad pathway in low‐dose Cd‐induced SDC4 reduction in HRGECs. Given that SDC4 is an essential component of glycocalyx, protection or repair of endothelial glycocalyx is a potential strategy for preventing or treating kidney diseases associated with environmental Cd exposure. This study aims to explore the effects of low‐dose cadmium on syndecan‐4 expression. The results revealed that Cd exposure decreases SDC4 expression in both mice kidney tissues and human renal glomerular endothelial cells (HRGECs). Cd exposure induces hyperpermeability in HRGECs and increases the levels P‐Smad2/3 and MMP‐2/9. TGF‐β signaling inhibitor SB431542 blocks Cd‐induced SDC4 reduction and MMP‐2/9 upregulation. The study revealed that low‐dose Cd reduces expression of SDC4 through activation of the TGF‐β signaling. [ABSTRACT FROM AUTHOR]

Tumors typically lack canonical danger signals required to activate adaptive immunity and also frequently employ substantial immunomodulatory mechanisms that downregulate adaptive responses and contribute to escape from immune surveillance. Given the variety of mechanisms involved in shielding tumors from immune recognition, it is not surprising that single‐agent immunomodulatory approaches have been largely unsuccessful in generating durable antitumor responses. Here we report a unique combination of immunomodulatory and cytostatic agents that recondition the tumor microenvironment and eliminate complex and/or poor‐prognosis tumor types including the non‐immunogenic 4T‐1 model of TNBC, the aggressive MOC‐2 model of HNSCC, and the high‐risk MYCN‐amplified model of neuroblastoma. A course of therapy optimized for TNBC cured a majority of tumors in both ectopic and orthotopic settings and eliminated metastatic spread in all animals tested at the highest doses. Immune responses were transferable between therapeutic donor and naïve recipient through adoptive transfer, and a sizeable abscopal effect on distant, untreated lesions could be demonstrated experimentally. Similar results were observed in HNSCC and neuroblastoma models, with characteristic remodeling of the tumor microenvironment documented in all model systems. scRNA‐seq analysis implicated upregulation of innate immune responses and antigen presentation in tumor cells and the myeloid cell compartment as critical early events. This analysis also highlighted the potential importance of the autonomic nervous system in the governance of inflammatory processes. The data indicate that the targeting of multiple pathways and mechanisms of action can result in substantial synergistic antitumor effects and suggest follow‐up in the neoadjuvant setting may be warranted. [ABSTRACT FROM AUTHOR]

Our previous studies have shown that upregulation of SLC7A1 in epithelial ovarian cancer (EOC) tumor cells significantly increases cancer cell proliferation, migration, and cisplatin resistance; however, the molecular mechanism by which SLC7A1 functions in EOC remains unknown. In later studies, we found that SLC7A1 is also highly expressed in the interstitial portion of high‐grade serous ovarian cancer (HGSOC), but the significance of this high expression in the interstitial remains unclear. Here, we showed the Interstitial high expression of SLC7A1 in HGSOC by immunohistochemistry. SLC7A1 enriched in cancer‐associated fibroblasts (CAFs) was upregulated by TGF‐β1. Transwell assay, scratch assay, cck8 assay and cell adhesion assay showed that SLC7A1 highly expressed in CAFs promoted tumor cells invasion, migration and metastasis in vitro. The effect of SLC7A1 on MAPK and EMT pathway proteins in ovarian cancer (OC) was verified by RNA sequencing and western blotting. Overexpression of SLC7A1 in OC is involved in MAPK/ ERK pathway and EMT. In general, in HGSOC, CAFs overexpressing SLC7A1 supported the migration and invasion of tumor cells; SLC7A1 is highly expressed in ovarian cancer and is involved in ERK phosphorylation and EMT signaling in MAPK signaling pathway. This suggests that SLC7A1 may be a potential therapeutic target for OC metastasis. [ABSTRACT FROM AUTHOR]

Cells integrate many mechanical and chemical cues to drive cell signalling responses. Because of the complex nature and interdependency of alterations in extracellular matrix (ECM) composition, ligand density, mechanics, and cellular responses it is difficult to tease out individual and combinatorial contributions of these various factors in driving cell behavior in homeostasis and disease. Tuning of material viscous and elastic properties, and ligand densities, in combinatorial fashions would enhance our understanding of how cells process complex signals. For example, it is known that increased ECM mechanics and transforming growth factor beta (TGF‐β) receptor (TGF‐β‐R) spacing/clustering independently drive TGF‐β signalling and associated myofibroblastic differentiation. However, it remains unknown how these inputs orthogonally contribute to cellular outcomes. Here, we describe the development of a novel material platform that combines microgel thin films with controllable viscoelastic properties and DNA origami to probe how viscoelastic properties and nanoscale spacing of TGF‐β‐Rs contribute to TGF‐β signalling and myofibroblastic differentiation. We found that highly viscous materials with non‐fixed TGF‐β‐R spacing promoted increased TGF‐β signalling and myofibroblastic differentiation. This is likely due to the ability of cells to better cluster receptors on these surfaces. These results provide insight into the contribution of substrate properties and receptor localisation on downstream signalling. Future studies allow for exploration into other receptor‐mediated processes. [ABSTRACT FROM AUTHOR]

Idiopathic pulmonary fibrosis (IPF) is a chronic lung condition characterized by the abnormal regulation of extracellular matrix (ECM) and epithelial‐mesenchymal transition (EMT). In this study, we investigated the potential of rutin, a natural flavonoid, in attenuating transforming growth factor‐β (TGF‐β)‐induced ECM regulation and EMT through the inhibition of the TGF‐β type I receptor (TβRI)‐mediated suppressor of mothers against decapentaplegic (SMAD) signaling pathway. We found that non‐toxic concentrations of rutin attenuated TGF‐β‐induced ECM‐related genes, including fibronectin, elastin, collagen 1 type 1, and TGF‐β, as well as myoblast differentiation from MRC‐5 lung fibroblast cells accompanied by the downregulation of α‐smooth muscle actin. Rutin also inhibited TGF‐β‐induced EMT processes, such as wound healing, migration, and invasion by regulating EMT‐related gene expression. Additionally, rutin attenuated bleomycin‐induced lung fibrosis in mice, thus providing a potential therapeutic option for IPF. The molecular docking analyses in this study predict that rutin occludes the active site of TβRI and inhibits SMAD‐mediated fibrotic signaling pathways in lung fibrosis. These findings highlight the potential of rutin as a promising anti‐fibrotic prodrug for lung fibrosis and other TGF‐β‐induced fibrotic and cancer‐related diseases; however, further studies are required to validate its safety and effectiveness in other experimental models. [ABSTRACT FROM AUTHOR]

Background: JAK inhibitors are well known for the treatment of rheumatoid arthritis (RA), but whether they can be used to treat pulmonary fibrosis, a common extra‐articular disease of RA, remains to be clarified. Methods: A jak2 inhibitor, CEP33779 (CEP), was administered to a rat model of RA‐associated interstitial lung disease to observe the degree of improvement in both joint swelling and pulmonary fibrosis. HFL1 cells were stimulated with TGF‐β1 to observe the expression of p‐JAK2. Then, different concentrations of related gene inhibitors (JAK2, TGFβ‐R1/2, and p‐STAT3) or silencers (STAT3, JAK2) were administered to HFL1 cells, and the expression levels of related proteins were detected to explore the underlying mechanisms of action. Results: CEP not only reduced the degree of joint swelling and inflammation in rats but also improved lung function, inhibited the pro‐inflammatory factors IL‐1β and IL‐6, reduced lung inflammation and collagen deposition, and alleviated lung fibrosis. CEP decreased the expression levels of TGFβ‐R2, p‐SMAD, p‐STAT3, and ECM proteins in rat lung tissues. TGF‐β1 induced HFL1 cells to highly express p‐JAK2, with the most pronounced expression at 48 h. The levels of p‐STAT3, p‐SMAD3, and ECM‐related proteins were significantly reduced after inhibition of either JAK2 or STAT3. Conclusion: JAK2 inhibitors may be an important and novel immunotherapeutic drug that can improve RA symptoms while also delaying or blocking the development of associated pulmonary fibrotic disease. The mechanism may be related to the downregulation of p‐STAT3 protein via inhibition of the JAK2/STAT signaling pathway, which affects the phosphorylation of SMAD3. [ABSTRACT FROM AUTHOR]

The Wiskott–Aldrich syndrome protein (WASp) regulates actin cytoskeletal dynamics and function of hematopoietic cells. Mutations in the WAS gene lead to two different syndromes; Wiskott–Aldrich syndrome (WAS) caused by loss‐of‐function mutations, and X‐linked neutropenia (XLN) caused by gain‐of‐function mutations. We previously showed that WASp‐deficient mice have a decreased number of regulatory T (Treg) cells in the thymus and the periphery. We here evaluated the impact of WASp mutations on Treg cells in the thymus of WAS and XLN mouse models. Using in vitro Treg differentiation assays, WAS CD4 single‐positive thymocytes have decreased differentiation to Treg cells, despite normal early signaling upon IL‐2 and TGF‐β stimulation. They failed to proliferate and express CD25 at high levels, leading to poor survival and a lower number of Foxp3+ Treg cells. Conversely, XLN CD4 single‐positive thymocytes efficiently differentiate into Foxp3+ Treg cells following a high proliferative response to IL‐2 and TGF‐β, associated with high CD25 expression when compared with WT cells. Altogether, these results show that specific mutations of WASp affect Treg cell development differently, demonstrating a critical role of WASp activity in supporting Treg cell development and expansion. [ABSTRACT FROM AUTHOR]

Background Information: The precise etiology of breast cancer is not completely understood, although women with BRCA1 gene mutations have a significantly increased risk of developing the disease. In addition, sporadic breast cancer is frequently associated with decreased BRCA1 gene expression. Growing evidence of Human papillomaviruses (HPVs) infections in breast tumors has raised the possibility of the involvement of HPVs in the pathogenesis of breast cancer. We investigated whether the effects of HPV oncoproteins E6 and E7 were influenced by the expression levels of BRCA1. HPV16E6E7 (prototype or E6D25E/E7N29S Asian variant type) were stably expressed in MDA‐MB231 breast cancer cells, wild type for BRCA1, or with BRCA1 knocked down. Results: Expression of HPV16E6E7 oncogenes did not affect BRCA1 levels and the abundance of HPV16E6E7 was not altered by BRCA1 knockdown. BRCA1 levels did not alter HPV16E6E7‐dependent degradation of G1‐S cell cycle proteins p53 and pRb. However, we found that the expression of G2‐M cell cycle protein cyclin B1 enhanced by HPV16E6E7 was impacted by BRCA1 levels. Especially, we found the correlation between BRCA1 and cyclin B1 expression and this was also confirmed in breast cancer samples from a Thai cohort. We further demonstrated that the combination of HPV oncoproteins and low levels of BRCA1 protein appears to enhance proliferation and invasion. Transactivation activities of HPV16E6E7 on genes regulating cell proliferation and invasion (TGF‐β and vimentin) were significantly increased in BRCA1‐deficient cells. Conclusions: Our results indicate that a deficiency of BRCA1 promotes the transactivation activity of HPV16E6E7 leading to increase of cell proliferation and invasion. Significance: HPV infection appears to have the potential to enhance the aggressiveness of breast cancers, especially those deficient in BRCA1. [ABSTRACT FROM AUTHOR]

Aims: The dysregulation of TGF‐β signaling is a crucial pathophysiological process in tumorigenesis and progression. LncRNAs have diverse biological functions and are significant participants in the regulation of tumor signaling pathways. However, the clinical value of lncRNAs related to TGF‐β signaling in glioma is currently unclear. Methods: Data on glioma's RNA‐seq transcriptome, somatic mutation, DNA methylation data, and clinicopathological information were derived from the CGGA and TCGA databases. A prognostic lncRNA signature was constructed by Cox and LASSO regression analyses. TIMER2.0 database was utilized to deduce immune infiltration characteristics. "ELMER v.2" was used to reconstruct TF‐methylation‐gene regulatory network. Immunotherapy and chemotherapy response predictions were implemented by the TIDE algorithm and GDSC database, respectively. In vitro and in vivo experiments were conducted to verify the results and clarify the regulatory mechanism of lncRNA. Results: In glioma, a TGF‐β signaling‐related 15‐lncRNA signature was constructed, including AC010173.1, HOXA‐AS2, AC074286.1, AL592424.1, DRAIC, HOXC13‐AS, AC007938.1, AC010729.1, AC013472.3, AC093895.1, AC131097.4, AL606970.4, HOXC‐AS1, AGAP2‐AS1, and AC002456.1. This signature proved to be a reliable prognostic tool, with high risk indicating an unfavorable prognosis and being linked to malignant clinicopathological and genomic mutation traits. Risk levels were associated with different immune infiltration landscapes, where high risk was indicative of high levels of macrophage infiltration. In addition, high risk also suggested better immunotherapy and chemotherapy response. cg05987823 was an important methylation site in glioma progression, and AP‐1 transcription factor family participated in the regulation of signature lncRNA expression. AGAP2‐AS1 knockdown in in vitro and in vivo experiments inhibited the proliferation, migration, and invasion of glioma cells, as well as the growth of glioma, by downregulating the expression levels of NF‐κB and ERK 1/2 in the TGF‐β signaling pathway. Conclusions: A prognostic lncRNA signature of TGF‐β signaling was established in glioma, which can be used for prognostic judgment, immune infiltration status inference, and immunotherapy response prediction. AGAP2‐AS1 plays an important role in glioma progression. [ABSTRACT FROM AUTHOR]

Aim: Cyst formation of the jaws is frequently accompanied by the proliferation of odontogenic epithelial cells located in the periodontal ligament (PDL), which consists of heterozygous cells and includes the most fibroblasts. The lining epithelium of radicular cyst, an odontogenic cyst of inflammatory origin, is derived from the proliferation of the remnants of the Hertwig epithelial root sheath (odontogenic epithelial cell rests of Malassez; ERMs) in the PDL. ERMs are maintained at a lower proliferative state under physiological conditions, but the regulatory mechanisms underlying the inflammation‐dependent enhanced‐proliferative capabilities of ERMs are not fully understood. The aim of this study was to evaluate the effects of cytokine pathway association between TGF‐β signalling and IL‐1β signalling on the regulation of odontogenic epithelial cell proliferation using radicular cyst pathological specimens and odontogenic epithelial cell lines. Methodology: Immunofluorescence analyses were performed to clarify the expression levels of Smad2/3 and Ki‐67 in ERMs of 8‐week‐old mouse molar specimens. In radicular cyst (n = 52) and dentigerous cysts (n = 6) specimens from human patients, the expression of p65 (a main subunit of NF‐κB), Smad2/3 and Ki‐67 were investigated using immunohistochemical analyses. Odontogenic epithelial cells and PDL fibroblastic cells were co‐cultured with or without an inhibitor or siRNAs. Odontogenic epithelial cells were cultured with or without TGF‐β1 and IL‐1β. The proliferative capabilities and Smad2 phosphorylation levels of odontogenic epithelial cells were examined. Results: Immunohistochemically, Smad2/3‐positivity was increased, and p65‐positivity and Ki‐67‐positivity were decreased both in ERMs and in the epithelial cells in dentigerous cysts, a non‐inflammatory developmental cyst. In contrast, p65‐positive cells, along with the expression of Ki‐67, were increased and Smad2/3‐positive cells were decreased in the lining epithelia of radicular cysts. Co‐culture experiments with odontogenic epithelial cells and PDL fibroblastic cells revealed that PDL cells‐derived TGF‐β1/2 and their downstream signalling suppressed odontogenic epithelial cell proliferation. Moreover, TGF‐β1 stimulation induced Smad2 phosphorylation and suppressed odontogenic epithelial cell proliferation, while IL‐1β stimulation reversed these phenotypes through p65 transactivation. Conclusions: These results suggest that IL‐1β‐p65 signalling promotes odontogenic epithelial cell proliferation through suppressing TGF‐β‐Smad2 signalling, which would be involved in the pathogenesis of radicular cysts. [ABSTRACT FROM AUTHOR]

Background: Wound healing has long been a subject of great interest for pathologists. Previous research has established the effectiveness of topical application of chitooligosaccharides (COSs) on wound healing. However, studies on their potential usefulness as food additives to improve wound healing are lacking. Objective: Here, we investigated whether dietary supplementation with COSs could promote wound healing using Beagle dogs as an animal model. Methods: Nine dogs were evenly assigned randomly into control, 0.5% COSs and 1% COSs groups. Control dogs were fed with a non‐supplemented dietary formula (vehicle). Dogs in 0.5% COSs and 1% COSs groups were fed with a dietary formula supplemented with 0.5% and 1% COSs. After 8 weeks of feeding, dogs in each group underwent surgery with three matching full‐thickness 2 × 2‐cm skin defects were created with a scalpel on the left side of the dorsal midline, 2 cm apart from each other. Wounds were photographed and the healing percentage was determined on day 0, 2, 4, 7, 9, 11, 14, 16, 18 and 21. The healing process was assessed by daily gross examination, histologic examination and Masson's trichrome staining on days 7, 14 and 21. Considering the important role of the transforming growth factor‐β (TGF‐β1) signalling pathway in wound healing, immunostaining was used to evaluate TGF‐β1 expression. Results: The results showed that wound healing was accelerated; collagen synthesis and TGF‐β1 expression were enhanced in response to dietary supplementation with COSs. Conclusions: This study makes an original contribution to the understanding of wound healing in dogs and other animals and supports the use of COSs as a supplement to be added to animal diets. [ABSTRACT FROM AUTHOR]

The processes involved in regeneration of cutaneous compared to corneal tissues involve different intrinsic mechanisms. Importantly, cutaneous wounds involve healing by angiogenesis but vascularization of the cornea obscures vision. Previous studies showed that topically applied calreticulin (CALR) healed full‐thickness excisional animal wounds by a tissue regenerative process markedly enhancing repair without evoking angiogenesis. In the current study, the application of CALR in a rabbit corneal injury model: (1) accelerated full wound closure by 3 days (2) accelerated delayed healing caused by corticosteroids, routinely used to prevent post‐injury inflammation, by 6 days and (3) healed wounds without vascularization or fibrosis/hazing. In vitro, CALR stimulated proliferation of human corneal epithelial cells (CE) and corneal stromal cells (keratocytes) by 1.5‐fold and 1.4‐fold, respectively and induced migration of CE cells and keratocytes, by 72% and 85% compared to controls of 44% and 59%, respectively. As a marker of decreased fibrosis, CALR treated corneal wounds showed decreased immunostaining for α‐smooth muscle actin (α‐SMA) by keratocytes and following CALR treatment in vitro, decreased the levels of TGF‐β2 in human CE cells and α‐SMA in keratocytes. CALR has the potential to be a novel therapeutic both, to accelerate corneal healing from various injuries and in conjunction with corticosteroids. [ABSTRACT FROM AUTHOR]

According to recent research, metabolic‐associated fatty liver disease (MAFLD) has emerged as an important underlying etiology of hepatocellular carcinoma (HCC). However, the molecular mechanism of MAFLD‐HCC is still unclear. Tumor necrosis factor receptor‐associated factor 2 (TRAF2) is the key molecule to mediate the signal of inflammatory NF‐κB pathway. This study aims to investigate the potential dysregulation of TRAF2 and its biological function in MAFLD‐HCC. Huh7 TRAF2−/− demonstrated increased tumor formation ability compared to huh7 TRAF2+/+ when stimulated with transforming growth factor‐β (TGF‐β). The decisive role of TGF‐β in the development of MAFLD‐HCC was confirmed through the specific depletion of TGF‐β receptor II gene in the hepatocytes (Tgfbr2ΔHep) of mice. In TRAF2−/− cells treated with TGF‐β, both the glycolysis rate and lipid synthesis were enhanced. We proved the signal of the mechanistic target of rapamycin complex 1 (mTORC1) could be activated in the presence of TGF‐β, and was enhanced in TRAF2−/− cells. The coimmunoprecipitation (co‐IP) experiments revealed that TRAF2 fortified the Smurf2‐mediated ubiquitination degradation of AXIN1. Hence, TRAF2 depletion resulted in increased Smad7 degradation induced by AXIN1, thus promoting the TGF‐β signal. We also discovered that PLX‐4720 could bind with AXIN1 and restrained the tumor proliferation of TRAF2−/− in mice fed with high‐fat diet (HFD). Our findings indicate that TRAF2 plays a significant role in the pathogenesis of MAFLD‐HCC. The reduction of TRAF2 expression leads to the enhancement of the TGF‐β‐mTORC1 pathway by facilitating AXIN1‐mediated Smad7 degradation. [ABSTRACT FROM AUTHOR]

Macrophages play a pivotal role in the response to injury, contributing significantly to the repair and regrowth of damaged tissues. The external lateral line system in aquatic organisms offers a practical model for studying regeneration, featuring interneuromast cells connecting sensory neuromasts. Under normal conditions, these cells remain dormant, but their transformation into neuromasts occurs when overcoming inhibitory signals from Schwann cells and posterior lateral line nerves. The mechanism enabling interneuromast cells to evade inhibition by Schwann cells remains unclear. Previous observations suggest that macrophages physically interact with neuromasts, nerves, and Schwann cells during regeneration. This interaction leads to the regeneration of neuromasts in a subset of zebrafish with ablated neuromasts. To explore whether macrophages achieve this effect through secreted cytokines, we conducted experiments involving tail amputation in zebrafish larvae and tested the impact of cytokine inhibitors on neuromast regeneration. Most injured larvae remarkably regenerated a neuromast within 4 days post‐amputation. Intriguingly, removal of macrophages and inhibition of the anti‐inflammatory cytokine transforming growth factor‐beta (TGF‐β) significantly delayed neuromast regeneration. Conversely, inhibition of the pro‐inflammatory cytokines interleukin‐6 (IL‐6) and tumor necrosis factor‐alpha (TNF‐α) had minor effects on the regeneration process. This study provides insights into how macrophages activate interneuromast cells, elucidating the pathways underlying neuromast regeneration. [ABSTRACT FROM AUTHOR]

TGF‐β is thought to be involved in the physiological functions of early organ development and pathological changes in substantial organ fibrosis, while studies around adipose tissue function and systemic disorders of glucolipid metabolism are still scarce. In this investigation, two animal models, aP2‐SREBP‐1c mice and ob/ob mice, were used. TGF‐β pathway showed up‐regulated in the inguinal white adipose tissue (iWAT) of the two models. SB431542, a TGF‐β inhibitor, successfully increased inguinal white adipocyte size by more than 1.5 times and decreased the weight of Peripheral organs including liver, Spleen and Kidney to 73.05%/62.18%/73.23% of pre‐administration weights. The iWAT showed elevated expression of GLUTs and lipases, followed by a recovery of circulation GLU, TG, NEFA, and GLYCEROL to the wild‐type levels in aP2‐SREBP‐1c mice. In contrast, TGF‐β inhibition did not have similar effects on that of ob/ob mice. In vitro, TGF‐β blocker treated mature adipocytes had considerably higher levels of glycerol and triglycerides than the control group, whereas GLUTs and lipases expression levels were unchanged. These findings show that inhibiting the abnormally upregulated TGF‐β pathway will only restore iWAT expansion and ameliorate the global metabolic malfunction of glucose and lipids in lipodystrophy, not obesity. [ABSTRACT FROM AUTHOR]

Physiological embryogenesis and adult tissue homeostasis are regulated by transforming growth factor‐β (TGF‐β), an evolutionarily conserved family of secreted polypeptide factors, acting in an autocrine and paracrine manner. The role of TGF‐β in inflammation, fibrosis, and cancer is complex and sometimes even contradictory, exhibiting either inhibitory or promoting effects depending on the stage of the disease. Under pathological conditions, especially fibrosis and cancer, overexpressed TGF‐β causes extracellular matrix deposition, epithelial–mesenchymal transition, cancer‐associated fibroblast formation, and/or angiogenesis. In this review article, we have tried to dive deep into the mechanism of action of TGF‐β in inflammation, fibrosis, and carcinogenesis. As TGF‐β and its downstream signaling mechanism are implicated in fibrosis and carcinogenesis blocking this signaling mechanism appears to be a promising avenue. However, targeting TGF‐β carries substantial risk as this pathway is implicated in multiple homeostatic processes and is also known to have tumor‐suppressor functions. There is a need for careful dosing of TGF‐β drugs for therapeutic use and patient selection. [ABSTRACT FROM AUTHOR]

Tumor progression and metastasis are regulated by endothelial cells undergoing endothelial–mesenchymal transition (EndoMT), a cellular differentiation process in which endothelial cells lose their properties and differentiate into mesenchymal cells. The cells undergoing EndoMT differentiate through a spectrum of intermediate phases, suggesting that some cells remain in a partial EndoMT state and exhibit an endothelial/mesenchymal phenotype. However, detailed analysis of partial EndoMT has been hampered by the lack of specific markers. Transforming growth factor‐β (TGF‐β) plays a central role in the induction of EndoMT. Here, we showed that inhibition of TGF‐β signaling suppressed EndoMT in a human oral cancer cell xenograft mouse model. By using genetic labeling of endothelial cell lineage, we also established a novel EndoMT reporter cell system, the EndoMT reporter endothelial cells (EMRECs), which allow visualization of sequential changes during TGF‐β‐induced EndoMT. Using EMRECs, we characterized the gene profiles of multiple EndoMT stages and identified CD40 as a novel partial EndoMT‐specific marker. CD40 expression was upregulated in the cells undergoing partial EndoMT, but decreased in the full EndoMT cells. Furthermore, single‐cell RNA sequencing analysis of human tumors revealed that CD40 expression was enriched in the population of cells expressing both endothelial and mesenchymal cell markers. Moreover, decreased expression of CD40 in EMRECs enhanced TGF‐β‐induced EndoMT, suggesting that CD40 expressed during partial EndoMT inhibits transition to full EndoMT. The present findings provide a better understanding of the mechanisms underlying TGF‐β‐induced EndoMT and will facilitate the development of novel therapeutic strategies targeting EndoMT‐driven cancer progression and metastasis. [ABSTRACT FROM AUTHOR]

Desmoid tumors (DTs), also called desmoid‐type fibromatoses, are locally aggressive tumors of mesenchymal origin. In the present study, we developed a novel mouse model of DTs by inducing a local mutation in the Ctnnb1 gene, encoding β‐catenin in PDGFRA‐positive stromal cells, by subcutaneous injection of 4‐hydroxy‐tamoxifen. Tumors in this model resembled histologically clinical samples from DT patients and showed strong phosphorylation of nuclear SMAD2. Knockout of SMAD4 in the model significantly suppressed tumor growth. Proteomic analysis revealed that SMAD4 knockout reduced the level of Cysteine‐and‐Glycine‐Rich Protein 2 (CSRP2) in DTs, and treatment of DT‐derived cells with a TGF‐β receptor inhibitor reduced CSRP2 RNA levels. Knockdown of CSRP2 in DT cells significantly suppressed their proliferation. These results indicate that the TGF‐β/CSRP2 axis is a potential therapeutic target for DTs downstream of TGF‐β signaling. [ABSTRACT FROM AUTHOR]

Osteoarthritis (OA) is a common chronic degenerative disease which is characterized by the disruption of articular cartilage. Syringic acid (SA) is a phenolic compound with anti‐inflammatory, antioxidant, and other effects including promoting osteogenesis. However, the effect of SA on OA has not yet been reported. Therefore, the purpose of our study was to investigate the effect and mechanism of SA on OA in a mouse model of medial meniscal destabilization. The expressions of genes were evaluated by qPCR or western blot or immunofluorescence. RNA‐seq analysis was performed to examine gene transcription alterations in chondrocytes treated with SA. The effect of SA on OA was evaluated using destabilization of the medial meniscus model of mice. We found that SA had no obvious toxic effect on chondrocytes, while promoting the expressions of chondrogenesis‐related marker genes. The results of RNA‐seq analysis showed that extracellular matrix–receptor interaction and transforming growth factor‐β (TGF‐β) signaling pathways were enriched among the up‐regulated genes by SA. Mechanistically, we demonstrated that SA transcriptionally activated Smad3. In addition, we found that SA inhibited the overproduction of lipopolysaccharide‐induced inflammation‐related cytokines including tumor necrosis factor‐α and interleukin‐1β, as well as matrix metalloproteinase 3 and matrix metalloproteinase 13. The cell apoptosis and nuclear factor‐kappa B (NF‐κB) signaling were also inhibited by SA treatment. Most importantly, SA attenuated cartilage degradation in a mouse OA model. Taken together, our study demonstrated that SA could alleviate cartilage degradation in OA by activating the TGF‐β/Smad and inhibiting NF‐κB signaling pathway. [ABSTRACT FROM AUTHOR]

Bone metastases are a common cause of suffering in breast and prostate cancer patients, however, the interaction between bone cells and cancer cells is poorly understood. Using a series of co‐culture, conditioned media, human cancer spheroid, and organ‐on‐a‐chip experiments, this study reveals that osteocytes suppress cancer cell proliferation and increase migration via tumor necrosis factor alpha (TNF‐α) secretion. This action is regulated by osteocyte primary cilia and associated intraflagellar transport protein 88 (IFT88). Furthermore, it shows that cancer cells block this mechanism by secreting transforming growth factor beta (TGF‐β), which disrupts osteocyte cilia and IFT88 gene expression. This bi‐directional crosstalk signaling between osteocytes and cancer cells is common to both breast and prostate cancer. This study also proposes that osteocyte inhibition of cancer cell proliferation decreases as cancer cells increase, producing more TGF‐β. Hence, a positive feedback loop develops accelerating metastatic tumor growth. These findings demonstrate the importance of cancer cell‐osteocyte signaling in regulating breast and prostate bone metastases and support the development of therapies targeting this pathway. [ABSTRACT FROM AUTHOR]

We previously reported that patients with multidrug‐resistant tuberculosis (MDR‐TB) showed low systemic and Mtb‐induced Th22 responses associated to high sputum bacillary load and severe lung lesions suggesting that Th22 response could influence the ability of these patients to control bacillary growth and tissue damage. In MDR‐TB patients, the percentage of IL‐22+ cells inversely correlates with the proportion of senescent PD‐1+ T cells. Herein, we aimed to evaluate the pathways involved on the regulation of systemic and Mtb‐induced Th22 response in MDR‐TB and fully drug‐susceptible TB patients (S‐TB) and healthy donors. Our results show that while IL‐1β and IL‐23 promote Mtb‐induced IL‐22 secretion and expansion of IL‐22+ cells, TGF‐β inhibits this response. Systemic and in vitro Mtb‐induced Th22 response inversely correlates with TGF‐β amounts in plasma and in PBMC cultures respectively. The number of circulating PD‐1+ T cells directly correlates with plasmatic TGF‐β levels and blockade of PD‐1/PD‐L1 signalling enhances in vitro Mtb‐induced expansion of IL‐22+ cells. Thus, TGF‐β could also inhibit Th22 response through upregulation of PD‐1 expression in T cells. Higher percentage of IL‐23+ monocytes was observed in TB patients. In contrast, the proportion of IL‐1β+ monocytes was lower in TB patients with bilateral lung cavities (BCC) compared to those patients with unilateral cavities (UCC). Interestingly, TB patients with BCC showed higher plasmatic and Mtb‐induced TGF‐β secretion than patients with UCC. Thus, high TGF‐β secretion and subtle differences in IL‐23 and IL‐1β expression could diminish systemic and in vitro Mtb‐induced Th22 response along disease progression in TB patients. [ABSTRACT FROM AUTHOR]

Background: Total flavonoids of Rhizoma drynariae (TFRD) is broadly used in the treatment of orthopedic diseases. Nevertheless, the effects and underlying mechanism of TFRD on tendon‐bone healing after anterior cruciate ligament reconstruction (ACLR) remain unclear. Methods: The ACLR mouse model was established. Hematoxylin and Eosin (HE) staining was used for histological analysis of tendon‐bone healing. Western blot was utilized to detect the levels of osteogenic related factors (ALP, OCN, RUNX2). The viability and alkaline phosphatase (ALP) activity of bone mesenchymal stem cells (BMSCs) were determined by Cell Counting Kit‐8 (CCK‐8) and ALP assays. The interaction of estrogen related receptor alpha (ESRRA), estrogen related receptor beta (ESRRB), and golgi‐localized γ‐ear containing ADP ribosylation factor‐binding protein 1 (Gga1) was detected by luciferase reporter assays. The levels of important proteins on the TGF‐β/MAPK pathway were measured by western blot. Results: TFRD improved tendon‐bone healing, restored biomechanics of ACLR mice and activated the TGF‐β/MAPK pathway. TFRD treatment also enhanced the viability and osteogenic differentiation of BMSCs in vitro. Then, we demonstrated that TFRD targeted ESRRA and ESRRB to transcriptionally activate Gga1 expression. Knockdown of ESRRA, ESRRB, or Gga1 suppressed the viability and osteogenic differentiation of TFRD‐induced BMSCs, which was revealed to be restored by Gga1 overexpression. The overexpression of ESRRA, ESRRB, or Gga1 was demonstrated to promote the BMSC viability and osteogenic differentiation. TGF‐β1 treatment can reverse the impact of Gga1 inhibition on osteogenic differentiation in TFRD‐induced BMSCs. Conclusion: TFRD improves tendon‐bone healing in ACLR mouse models and facilitates the osteogenic differentiation of BMSCs through the ERR1/2‐Gga1‐TGF‐β/MAPK pathway, which might deepen our understanding of the underlying mechanism of TFRD in tendon‐bone healing. [ABSTRACT FROM AUTHOR]

This article discusses the factors that determine the differentiation of autoreactive thymocytes into regulatory T cells (Tregs). Tregs are a subset of CD4+ T cells that play a crucial role in maintaining peripheral tolerance and immune homeostasis. The majority of Tregs are generated in the thymus upon recognition of self-antigens, while others can be generated in specialized peripheral tissues. The differentiation of Tregs is influenced by factors such as T-cell receptor signaling, co-stimulation, and the presence of TGF-β and retinoic acid. The study also highlights the role of late agonist signaling and the disruption of TCR signaling in the generation of Tregs. The findings contribute to our understanding of thymocyte selection and central tolerance, and may have implications for future therapeutic interventions involving Tregs. [Extracted from the article]

In this study, we found that in the adipose tissue of wildtype animals, insulin and TGF‐β signalling converge via a BMP antagonist short gastrulation (sog) to regulate ECM remodelling. In tumour bearing animals, Sog also modulates TGF‐β signalling to regulate ECM accumulation in the fat body. TGF‐β signalling causes ECM retention in the fat body and subsequently depletes muscles of fat body‐derived ECM proteins. Activation of insulin signalling, inhibition of TGF‐β signalling, or modulation of ECM levels via SPARC, Rab10 or Collagen IV in the fat body, is able to rescue tissue wasting in the presence of tumour. Together, our study highlights the importance of adipose ECM remodelling in the context of cancer cachexia. Synopsis: During cancer cachexia, insulin and TGF‐β signalling converge in the fat body via Sog to hinder ECM secretion, which in turn affects muscle integrity.Eye tumour models employing RasV12 together with Scrib RNAi or Dlg RNAi were used to study how inter‐organ communication affects fat body and muscle integrity in cancer cachexia.TGF‐β and insulin signalling converge in the fat body to control ECM secretion via Sog.Modulating ECM secretion from the fat body rescues muscle integrity in the presence of tumour. [ABSTRACT FROM AUTHOR]

Numerous studies revealed that Chlorella vulgaris and orange peels are potential sources for many valuable compounds such as flavonoids, which are natural polyphenols with antioxidant capacities that lessen oxidative stress via suppressing ROS levels. Thus, this study was designed to investigate their radioprotective efficiency either alone or in combination as natural food supplements. Sixty‐four male Albino rats were divided into eight groups (n = 8) as follows: control, orange peel (10% in diet), C. vulgaris (1% in diet), orange peel + C. vulgaris, gamma irradiated (2Gy twice per week up to 8Gy), orange peel + gamma irradiation, C. vulgaris + gamma irradiation, and orange peel + C. vulgaris + gamma irradiation. After the experiment, blood serums were collected for biochemical analysis, whole bloods were collected for blood picture, bone marrows were collected for GSH, MDA, TGF‐β, NOX2 and NOX4, and liver tissues were collected for histopathological evaluation. Current study revealed that exposure to gamma irradiation induced a significant disturbance in liver function markers (ALT and AST), kidney function markers (urea and creatinine), cholesterol and triglycerides levels in serum. In addition, a significant decrease in WBCs, RBCs, PLT, and Hb in blood of irradiated rats. Moreover, a significant elevation in TGF‐β, NOX2, NOX4 activities, and MDA level, while showed a marked decrease in GSH concentration. Furthermore, hepatic inflammation appeared in the histopathological examination. Orange peels or C. vulgaris treatments showed acceptable amelioration in all measured parameters, combination between orange peels and C. vulgaris showed statistically significant additive amelioration in radiation induced disturbance. Significance statement: This study showed that orange peels and Chlorella vulgaris either alone or in combination exhibited acceptable amelioration in all measured parameters revealing their radioprotective capability. [ABSTRACT FROM AUTHOR]

Podocytes, the key component of the glomerular filtration barrier (GFB), are gradually lost during the progression of diabetic kidney disease (DKD), severely compromising kidney functionality. The molecular mechanisms regulating the survival of podocytes in DKD are incompletely understood. Here, we show that membrane-associated guanylate kinase inverted 2 (MAGI2) is specifically expressed in renal podocytes, and promotes podocyte survival in DKD. We found that MAGI2 expression was downregulated in podocytes cultured with high-glucose in vitro, and in kidneys of db/db mice as well as DKD patients. Conversely, we found enforced expression of MAGI2 via AAV transduction protected podocytes from apoptosis, with concomitant improvement of renal functions. Mechanistically, we found that MAGI2 deficiency induced by high glucose levels activates TGF-β signaling to decrease the expression of anti-apoptotic proteins. These results indicate that MAGI2 protects podocytes from cell death, and can be harnessed therapeutically to improve renal function in diabetic kidney disease. [ABSTRACT FROM AUTHOR]

Hepcidin negatively regulates systemic iron levels by inhibiting iron entry into the circulation. Hepcidin production is increased in response to an increase in systemic iron via the activation of the bone morphogenetic protein (BMP) pathway. Regulation of hepcidin expression by iron status has been proposed on the basis of evidence mainly from rodents and humans. We evaluated the effect of iron administration on plasma hepcidin concentrations in calves and the expression of bovine hepcidin by the BMP pathway in a cell culture study. Hematocrit as well as levels of blood hemoglobin and plasma iron were lower than the reference level in calves aged 1–4 weeks. Although intramuscular administration of iron increased iron‐related parameters, plasma hepcidin concentrations were unaffected. Treatment with BMP6 increased hepcidin expression in human liver‐derived cells but not in bovine liver‐derived cells. A luciferase‐based reporter assay revealed that Smad4 was required for hepcidin reporter transcription induced by Smad1. The reporter activity of hepcidin was lower in the cells transfected with bovine Smad4 than in those transfected with murine Smad4. The lower expression levels of bovine Smad4 were responsible for the lower activity of the hepcidin reporter, which might be due to the instability of bovine Smad4 mRNA. In fact, the endogenous Smad4 protein levels were lower in bovine cells than in human and murine cells. Smad4 also confers TGF‐β/activin‐mediated signaling. Induction of TGF‐β‐responsive genes was also lower after treatment with TGF‐β1 in bovine hepatocytes than in human hepatoma cells. We revealed the unique regulation of bovine hepcidin expression and the characteristic TGF‐β family signaling mediated by bovine Smad4. The present study suggests that knowledge of the regulatory expression of hepcidin as well as TGF‐β family signaling obtained in murine and human cells is not always applicable to bovine cells. [ABSTRACT FROM AUTHOR]

Toxocara canis is a global zoonosis infection that can cause chronic and long‐term toxocariasis in their paratenic host. The excretory‐secretory (ES) products of T. canis larvae are considered to be responsible for the Th2 polarization and regulatory immune responses in toxocariasis. The C‐type lectin family is one of the most prominent components of ES products of T. canis infective larvae. This study aimed to investigate the ameliorative effect of a T. canis C‐type lectin recombinant protein (rCTL), on experimental autoimmune encephalomyelitis (EAE) which is a T‐cell‐mediated autoimmune disease of the central nervous system. C57BL/6 mice were subcutaneously treated with 30 μg rCTL, three times at an interval of 1 week. EAE was induced by myelin oligodendrocyte glycoprotein 35–55 peptide (MOG35‐55 peptide) immunization, and weight and clinical scores were evaluated. Real time polymerase chain reaction was performed to evaluate the expression levels of T‐bet, Gata3, and Foxp3 in splenocytes. In addition, the levels of interleukin 4, interferon gamma, and tumour growth factor‐β (TGF‐β) were quantified by enzyme‐linked immunosorbent assay in splenocyte culture supernatants. The results indicated that the rCTL decreased clinical disability scores and delayed the onset of EAE. Furthermore, the data showed that rCTL treatment modulated the immune response, which was associated with upregulation of the mRNA expression of the Foxp3 gene and higher production of TGF‐β in rCTL‐treated mice. This study demonstrated that rCTL might be a potential agent to ameliorate EAE symptoms by stimulating anti‐inflammatory responses. [ABSTRACT FROM AUTHOR]

Non‐small cell lung cancer (NSCLC) accounts for ~85% of all lung cancer cases. Neferine is used as a traditional Chinese medicine with many pharmacological effects, including antitumor properties; however, it has not been reported whether neferine plays an anticancer role by causing pyroptosis in NSCLC cells. We used two typical lung cancer cell lines, A549 and H1299, and 42 lung cancer tissue samples to investigate the regulatory effects of neferine on TGF‐β and MST1. We also treated lung cancer cells with different concentrations of neferine to study its effects on lung cancer cell survival, migration, invasion, and epithelial–mesenchymal transition (EMT) as well as on pyroptosis. Lentivirus‐mediated gain‐of‐function studies of TGF‐β and MST1 were applied to validate the roles of TGF‐β and MST1 in lung cancer. Next, we used murine transplanted tumor models to evaluate the effect of neferine treatment on the metastatic capacity of lung cancer tissues. With increasing neferine concentration, the viability, migration, invasion, and EMT capacity of A549 and H1299 cells decreased, whereas pyroptosis increased. Neferine repressed TGF‐β expression to modulate the induction of reactive oxygen species (ROS) by MST1. Overexpression of TGF‐β in either in vitro or mouse‐transplanted A549 cells restored the inhibitory effect of neferine on tumor development. Overexpression of MST1 clearly enhanced pyroptosis. Neferine contributed to pyroptosis by regulating MST1 expression through downregulation of TGF‐β to induce ROS formation. Therefore, our study shows that neferine can serve as an adjuvant therapy for NSCLC patients. [ABSTRACT FROM AUTHOR]

Sesamol, a lignan obtained from roasted seeds of Sesamum indicum, has high antioxidant and anti‐inflammatory activity. In this study, we have investigated the effect of sesamol on Bleomycin (BLM) induced pulmonary toxicity as well as fibrosis in Wistar rats. Lung toxicity was induced by administration of BLM, 0.015 U/g ip, twice weekly for 28 days whereas lung fibrosis was induced by BLM, 0.015 U/g ip, every 5th day for 49 days. Sesamol administration was started 7 days before first dose of BLM in both the models. It was observed that sesamol 50 mg/kg most effectively attenuated pulmonary toxicity by reducing oxidative stress, inflammation and apoptosis. This dose was further evaluated for its anti‐fibrotic effect. It was observed that there was a significant reduction in fibrosis. Lung collagen content was markedly reduced. Furthermore, expression of pro‐fibrotic proteins, TGF‐β/SMAD and α‐SMA, was reduced and that of anti‐fibrotic protein, AMPK, was markedly increased. Even though the combination of sesamol with pirfenidone exhibited no additional protection than either drug alone, it is evident from our study that our test drug, sesamol is comparable in efficacy to pirfenidone. Thus, sesamol has promising therapeutic potential in treatment of pulmonary toxicity and fibrosis. [ABSTRACT FROM AUTHOR]

For targeted intervention in coronavirus disease 2019 (COVID‐19), there is a high medical need for biomarkers that predict disease progression and severity in the first days after symptom onset. This study assessed the utility of early transforming growth factor β (TGF‐β) serum levels in COVID‐19 patients to predict disease severity, fatality, and response to dexamethasone therapy. Patients with severe COVID‐19 had significantly higher TGF‐β levels (416 pg/mL) as compared to patients with mild (165 pg/mL, p < 0.0001) or moderate COVID‐19 (241 pg/mL; p < 0.0001). Receiver operating characteristics area under the curve values were 0.92 (95% confidence interval [CI] 0.85–0.99, cut‐off: 255 pg/mL) for mild versus severe COVID‐19, and 0.83 (95% CI 0.65–1.0, cut‐off: 202 pg/mL) for moderate versus severe COVID‐19. Patients who died of severe COVID‐19 had significantly higher TGF‐β levels (453 pg/mL) as compared to convalescent patients (344 pg/mL), and TGF‐β levels predicted fatality (area under the curve: 0.75, 95% CI 0.53–0.96). TGF‐β was significantly reduced in severely ill patients treated with dexamethasone (301 pg/mL) as compared to untreated patients (416 pg/mL; p < 0.05). Early TGF‐β serum levels in COVID‐19 patients predict, with high accuracy, disease severity, and fatality. In addition, TGF‐β serves as a specific biomarker to assess response to dexamethasone treatment. [ABSTRACT FROM AUTHOR]

Intrinsic drug resistance mechanisms of tumor cells often reduce intracellular drug concentration to suboptimal levels. Epithelial‐to‐mesenchymal transition (EMT) is a pivotal process in tumor progression and metastasis that confers an aggressive phenotype as well as resistance to chemotherapeutics. Therefore, it is imperative to develop novel strategies and identify new targets to improve the overall efficacy of cancer treatment. We developed SN38 (active metabolite of irinotecan)‐assembled glycol chitosan nanoparticles (cSN38) for the treatment of pancreatic ductal adenocarcinoma (PDAC). Furthermore, cSN38 and the TGF‐β1 inhibitor LY364947 formed composite nanoparticles upon self‐assembly (cSN38 + LY), which obviated the poor aqueous solubility of LY364947 and enhanced drug sensitivity. The therapeutic efficacy of cSN38 + LY nanotherapeutics was studied in vitro and in vivo using suitable models. The cSN38 nanoparticles exhibited an antitumor effect that was significantly attenuated by TGF‐β‐induced EMT. The cellular uptake of SN38 was impeded during EMT, which affected the therapeutic efficacy. The combination of LY364947 and cSN38 markedly enhanced the cellular uptake of SN38, increased cytotoxic effects, and inhibited EMT in PDAC cells in vitro. Furthermore, cSN38 + LY significantly inhibited PDAC xenograft growth in vivo. The cSN38 + LY nanoparticles increased the therapeutic efficacy of cSN38 via repressing the EMT of PDAC cells. Our findings provide a rationale for designing nanoscale therapeutics to combat PDAC. [ABSTRACT FROM AUTHOR]

Idiopathic pulmonary fibrosis (IPF) is a fatal interstitial lung disease characterized by chronic, progressive, and fibrotic lung injury. Although remarkable progress has been made toward understanding the pathogenesis of PF, finding more effective treatments for this fatal disease remains a challenge. In this study, we describe an innovative macrophage‐based approach to deliver anti‐fibrotic protein to the lung and inhibit PF in a mouse model of bleomycin (BLM)‐induced lung injury. We engineered macrophages to continuously secrete three types of proteins: interleukin‐10, which prevents inflammation; TGFRcFc, a soluble truncated TGF‐βR2 that blocks TGF‐β; and CD147, which induces matrix metalloproteinases (MMPs) and causes collagen degradation. Infusing these engineered macrophages into the lungs of BLM‐induced PF mouse models in an optimal pattern significantly ameliorated PF in mice. Specifically, the most effective therapeutic outcome was achieved by infusing IL‐10‐secreting macrophages on day 1, followed by TGFRcFc‐secreting macrophages on day 7 and CD147‐secreting macrophages on day 14 into the same mice after BLM treatment. Our data suggest that macrophage‐based delivery of anti‐fibrotic proteins to the lungs is a promising therapy for fibrotic lung disorders. [ABSTRACT FROM AUTHOR]

The prognosis of multiple myeloma (MM) patients combined with renal insufficiency is poor. Renal fibrosis is an important pathological cause for MM patients combined with renal insufficiency. It is reported that epithelial–mesenchymal transition (EMT) of renal proximal tubular epithelial cells is an important mechanism in renal fibrosis. We speculated that EMT might play an important role in the renal insufficiency of MM with unclear mechanism. MM cells derived exosomes could affect the function of targeted cells by delivering microRNAs (miRNAs). Literature has shown that the expression of miR‐21 is closely related to EMT. In this research, we found that co‐culture of HK‐2 cells (human renal proximal tubular epithelial cells) and exosomes derived from MM cells promoted the EMT of HK‐2 cells, resulting in the down‐regulation of epithelial‐related marker (E‐cadherin), and up‐regulation of stroma‐related marker (Vimentin). Meanwhile, the expression of SMAD7, one of the downstream targets in the TGF‐β signalling pathway, was suppressed and the expression of TGF‐β was increased. After transfecting the inhibitor of miR‐21 in MM cells, the expression of miR‐21 in exosomes secreted by MM cells was significantly decreased, and the co‐culture of these treated exosomes and HK‐2 cells inhibited the EMT of HK‐2 cells. In conclusion, these findings showed that exosomal miR‐21 derived from MM cells could promote renal EMT through targeting TGF‐β/SMAD7 signalling pathway. [ABSTRACT FROM AUTHOR]

Background and Purpose: MicroRNA‐9 (miR‐9) has previously been described as a dual‐functional RNA during breast cancer progression and its roles need to be clarified thoroughly. Experimental Approach: A miR‐9 knockout mode of mouse breast cancer, the MMTV‐PyMT model (PyMT‐miR‐9−/−), combined with different human breast cancer cell lines were used to evaluate the effects of miR‐9 on breast cancer initiation, progression and metastasis. Lin‐NECs (Neoplastic mammary epithelial cells) and pNECs (Pre‐neoplastic mammary epithelial cells) were isolated and subjected to tumour‐initiation assay. Whole‐mount staining of mammary gland and histology was performed to determine mammary gland growth. Tumour‐initiating analysis combining a series of in vitro experiments were carried out to evaluate miR‐9 roles in tumour‐initiating ability. RNA‐sequencing of human breast cancer cells, and mammary glands at hyperplastic stages and established tumours in PyMT and PyMT‐miR‐9−/− mice, ChIP and luciferase report assays were conducted to reveal the underlying mechanisms. Key Results: MiR‐9 is ectopically expressed in breast cancer and its level is negatively correlated with the prognosis, especially in basal‐like breast cancer patients. Additionally, miR‐9 is essential for breast cancer progression by promoting the expansion and activity of tumour‐initiating cells (TICs) in preneoplastic glands, established tumours and xenograft modes. Mechanistically, the activity of TICs hinges on a positive TGF‐β/miR‐9 regulatory loop mediated by the STARD13/YAP axis. Conclusions and Implications: These findings demonstrate that miR‐9 is an oncogenic miRNA rather than a tumour‐suppressor in breast cancer, calling for rectification of the model for this conserved and highly abundant miRNA. [ABSTRACT FROM AUTHOR]

Liver fibrosis is caused by chronic liver injury. There are limited treatments for it, and the pathogenesis is unclear. Therefore, there is an urgent need to explore the pathogenesis of liver fibrosis, and to try to identify new potential therapeutic targets. For this study we used the carbon tetrachloride abdominal injection induced liver fibrosis animal model in mice. Primary hepatic stellate cell isolation was performed by a density‐gradient separation method, and this was followed by immunofluorescence stain analyses. Signal pathway analysis was performed by dual‐luciferase reporter assay and western blotting. Our results showed that RUNX1 was upregulated in cirrhotic liver tissues compared with normal liver tissues. Besides, overexpression of RUNX1 caused more severe liver fibrosis lesions than control group under CCl4‐induced conditions. Moreover, α‐SMA expression in the RUNX1 overexpression group was significantly higher than in the control group. Interestingly, we found that RUNX1 could promote the activation of TGF‐β/Smads in a dual‐luciferase reporter assay. Thus we demonstrated that RUNX1 could be considered as a new regulator of hepatic fibrosis by activating TGF‐β/Smads signalling. Based on this, we concluded that RUNX1 may be developed as a new therapeutic target in the treatment of liver fibrosis in the future. In addition, this study also provides a new insight about the aetiology of liver fibrosis. [ABSTRACT FROM AUTHOR]

Aim: Methicillin‐resistant Staphylococcus aureus (MRSA) is a pathogenic bacterium that causes severe skin diseases with ulceration. Since MRSA are not susceptible to multiple antibiotics, novel therapeutic agents are desired. Jumihaidokuto, a combination of 10 crude drugs, is a Japanese traditional Kampo medicine that is used for the treatment of skin diseases. In this study, we investigated the effects of jumihaidokuto extract (JHT) on MRSA‐infected mice. Methods: Mice infected subcutaneously with MRSA FPR3757 were administered JHT orally to analyze mouse survival, the condition of the mouse skin ulcer area, and transforming growth factor (TGF)‐β levels in the ulcer tissue. To evaluate the role of crude drug components, modified JHTs were prepared by removing one crude drug component at a time from the formula and the same experiment was conducted. Results: JHT treatment significantly increased the survival rate. Ulcerated areas on the skin of MRSA‐infected mice were observed to be significantly reduced in size following JHT treatment. Furthermore, MRSA was not detected at the site of the ulcer in the JHT‐treated group. Additionally, TGF‐β levels decreased in the mouse skin ulcer tissues on treatment with JHT. All the crude drug components in JHT were found to partially contribute to the survival rate and healing of skin ulcers. Among the 10 crude drugs, Bupleurum root, cherry bark, Aralia rhizome, ginger, and Schizonepeta spike exhibited higher contribution than the other crude drugs. Conclusion: Our findings indicate the efficacy of JHT in promoting the healing of MRSA‐induced murine skin ulcers. [ABSTRACT FROM AUTHOR]

Background: Lung adenocarcinoma (LUAD) is highly malignant and associated with poor prognoses in patients worldwide. There has been widespread recognition that lncRNAs are tightly linked to LUAD tumorigenesis and development. Here, we identified that the LINC00621 level was increased in LUAD tissues and concerned with the poor prognoses in LUAD patients. Methods: Bioinformatical analysis and RT‐qPCR determined the level of LINC00621 in LUAD tissues and cell lines. The admeasurement of the proliferation, migration, and invasion abilities of LUAD cells was utilized in the CCK8 and Transwell formulas. Luciferase reporter assay was used to corroborate the downstream target genes of LINC00621. The phosphorylated SMAD3 protein was tested by Western blotting assay. The impression of LINC00621 knockdown on LUAD tumor growth and metastasis put into effect by murine models. ChIP‐qPCR assay was carried out to verify the transcriptional regulation by FOXA1 on LINC00621. Results: In vitro, the knockdown of LINC00621 significantly reduced the proliferative, migrating, and invasive abilities, the same was true for tumorigenesis and metastasis in vivo. MiR‐34a‐5p as a straight target of LINC00621 was ascertained, and LUAD patients with inferior miR‐34a‐5p levels had undesirable prognoses. Furthermore, TGFBR1 is an immediate and functional connection site of miR‐34a‐5p. Collectively, LINC00621 can sponge miR‐34a‐5p and upregulate TGFBR1 levels, which further sensitized TGF‐β signaling pathway. Finally, it was revealed that FOXA1 transcriptionally upregulated LINC00621. Conclusion: This study uncovered that FOXA1‐induced LINC00621 promotes LUAD progression via the miR‐34a‐5p/TGFBR1/TGF‐β axis, and is one novel therapeutic target that may be used in LUAD treatment. [ABSTRACT FROM AUTHOR]

Pancreatic ductal adenocarcinoma (PDAC) is marked by molecular heterogeneity and poor prognosis. Among the stemness‐related transcription factors, Spalt‐like Transcription Factor 4 (SALL4) is correlated with unfavorable outcomes; however, its roles in PDAC remain unclear. SALL4high expression defines a PDAC subpopulation characterized by a shortened patient survival. Although SALL4 expression was mostly evaluated in tumor cells, our findings identify this embryonic transcription factor as a new biomarker in PDAC‐derived stroma. Gene expression analysis reveals that the SALL4high PDAC subset is enriched in cancer stem cell properties and stromal enrichment pathways; notably, an interaction with cancer‐associated fibroblasts (CAF) activated by TGF‐β. A particular oncogenic network was unraveled where Netrin‐1 and TGF‐β1 collaborate to induce SALL4 expression in CAF and drive their cancer‐stemness‐promoting functions. A 7‐gene stromal signature related to SALL4high PDAC samples was highlighted and validated by immunochemistry for prognosis and clinical application. This SALL4‐related stroma discriminated pancreatic preinvasive from invasive lesions and was enriched in short‐term survivors. Our results show that SALL4 transcriptional activity controls a molecular network defined by a specific stromal signature that characterizes PDAC invasiveness and worse clinical outcomes. [ABSTRACT FROM AUTHOR]

Severe fever with thrombocytopenia syndrome virus (SFTSV) and severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) can cause the hyperproduction of inflammatory cytokines, which have pathological effects in patient including severe or fatal cytokine storms. To characterize the effect of SFTSV and SARS‐CoV‐2 infection on the production of cytokines in severe fever with thrombocytopenia syndrome (SFTS) and COVID‐19 patients, we performed an analysis of cytokines in SFTS and COVID‐19 patients and also investigated the role of interleukin‐10 (IL‐10) in vitro studies: lipopolysaccharide‐induced THP‐1‐derived macrophages, SFTSV infection of THP‐1 cells, and SARS‐CoV‐2 infection of THP‐1 cells. In this study, we found that levels of both IL‐10 and IL‐6 were significantly elevated, the level of transforming growth factor‐β (TGF‐β) was significantly decreased and IL‐10 was elevated earlier than IL‐6 in severe and critical COVID‐19 and fatal SFTS patients, and inhibition of IL‐10 signaling decreased the production of IL‐6 and elevated that of TGF‐β. Therefore, the hyperproduction of IL‐10 and IL‐6 and the low production of TGF‐β have been linked to cytokine storm‐induced mortality in fatal SFTS and severe and critically ill COVID‐19 patients and that IL‐10 can play an important role in the host immune response to severe and critical SARS‐CoV‐2 and fatal SFTSV infection. [ABSTRACT FROM AUTHOR]

Fibrosis is an important phenomenon as it can occur early in the pathogenesis of asthma; it may be associated with disease severity and resistance to therapy. There is a strong evidence that infection caused by human rhinovirus (HRV) contributes to remodelling process, but there is lack of studies clearly explaining this pathway. Synthetic peroxisome proliferator‐activated receptor (PPAR) γ presents immunomodulatory and anti‐inflammatory features. In this study, we examined immunomodulatory properties of ciglitazone – PPAR‐γ agonist, in development and modulation of airway remodelling. Epithelial cells (NHBE) and two lines of fibroblasts (WI‐38, HFL1) were stimulated with ciglitazone and rhinovirus. The expression of genes related to airway remodelling process were analysed in the cells; moreover NF‐κB, c‐Myc and STAT3 were silenced in order to estimate potential pathways involved. Ciglitazone decreased mRNA expression of MMP‐9 and TGF‐β. It also modified the expression of α‐SMA and collagen after rhinovirus infection. Transcription factors knockdown altered the levels of expression. The results suggest possible anti‐fibrotic activity of PPAR‐γ agonist in human airway cells. Ciglitazone has been shown to be dependent on NF‐κB‐ and STAT3‐related pathways, thus, the PPAR‐γ agonist may have therapeutic potential for the treatment of airway remodelling in asthma. [ABSTRACT FROM AUTHOR]

Introduction: The implications of infiltrative compared to non‐infiltrative growth of cutaneous basal cell carcinoma (BCC) on the tumor stroma and immune cell landscape are unknown. This is of clinical importance, because infiltrative BCCs, in contrast to other BCC subtypes, are more likely to relapse after surgery and radiotherapy. Materials and Methods: This descriptive cross‐sectional study analyzed 38 BCCs collected from 2018 to 2021. In the first cohort (n = 28), immune cells were characterized by immunohistochemistry and multiplex immunofluorescence staining for CD3, CD8, CD68, Foxp3, and α‐SMA protein expression. In the second cohort (n = 10) with matched characteristics (age, sex, location, and BCC subtype), inflammatory parameters, including TGF‐β1, TGF‐β2, ACTA2, IL‐10, IL‐12A, and Foxp3, were quantified via RT‐qPCR after isolating mRNA from BCC tissue samples and perilesional skin. Results: Infiltrative BCCs showed significantly increased levels of α‐SMA expression in fibroblasts (p = 0.0001) and higher levels of Foxp3+ (p = 0.0023) and CD3+ (p = 0.0443) T‐cells compared to non‐infiltrative BCCs. CD3+ (p = 0.0171) and regulatory T‐cells (p = 0.0026) were significantly increased in α‐SMA‐positive tumor stroma, whereas CD8+ T‐cells (p = 0.1329) and CD68+ myeloid cells (p = 0.2337) were not affected. TGF‐β1 and TGF‐β2 correlated significantly with ACTA2/α‐SMA mRNA expression (p = 0.020, p = 0.005). Conclusion: Infiltrative growth of BCCs shows a myofibroblastic stroma differentiation and is accompanied by an immunosuppressive tumor microenvironment. [ABSTRACT FROM AUTHOR]

Background: Lymphocyte antigen 6 complex locus K (LY6K), a glycosylphosphatidylinositol‐anchored protein, plays a dynamic role in cancer metastasis. In the current study, we deciphered the effects of LY6K on transforming growth factor‐β (TGF‐β) and epidermal growth factor (EGF) signaling through clathrin‐ and caveolin‐1 (CAV‐1)‐mediated endocytosis. Methods: Analysis of the TCGA and GTEx dataset were performed to explore the expression and survival of LY6K in cancer patients. Short interfering RNA (siRNA) was used to knockdown the expression of LY6K in human cervical cancer patients. The effect of lack of LY6K on cell proliferation, migration, and invasion was performed, and RT‐qPCR and immunoblotting were performed to identify LY6K‐affected TGF‐β and EGF signaling pathways. Additionally, Immunofluorescence (IF) and transmission electron microscope (TEM) were performed to identify the role of LY6K in CAV‐1‐ and Clathrin‐mediated endocytosis. Results: Lymphocyte antigen 6 complex locus K expression level is elevated in higher grade cervical cancer patients correlating with poor overall survival, progression‐free survival, and disease‐free survival. LY6K‐depletion in HeLa and SiHa cancer cells suppressed EGF‐induced proliferation and TGF‐β‐enhanced migration and invasion. Both TGF‐β receptor‐I (TβRI) and EGF receptor (EGFR) localized at the plasma membrane regardless of LY6K expression, and LY6K bound TβRI irrespective of the presence of TGF‐β; however, LY6K did not bind EGFR. LY6K‐depleted cells showed impaired Smad2 phosphorylation upon TGF‐β treatment and lower proliferation rates following long‐term treatment with EGF. We revealed the atypical movement of TβRI and EGFR from plasma membrane upon ligand stimulation in LY6K‐depleted cells and an impaired movement of the endocytic proteins clathrin and CAV‐1. Conclusions: Our study demonstrates the key role of LY6K in both clathrin‐ and CAV‐1‐mediated endocytic pathways regulated by TGF‐β and EGF, and it suggests a correlation between LY6K overexpression in cervical cancer cells and poor overall survival. [ABSTRACT FROM AUTHOR]

Septic arthritis is a joint disease caused by Staphylococcus aureus. Different macrophage populations contribute in various ways to control blood‐borne infections and induce inflammatory responses. Macrophage tissue–resident niche is necessary for the suppression of chronic inflammation and may contribute to the pathogenesis of septic arthritis. Thus, to obtain a resolution of the disease and restoration of synovial homeostasis, it needs the activation of macrophages that further regulate the inflammatory consequences. The aim of this study was to find out the mechanism by which neutralization of transforming growth factor‐beta (TGF‐β) and/or interleukin (IL)‐6 after induction of septic arthritis could alter the specific macrophage responses in spleen and synovial joints via different cytokines (osteoprotegerin (OPG), osteopontin (OPN), IL‐10, IL‐12 and CXCL8) cross‐talking, and how the response could be modulated by reactive oxygen species vs antioxidant enzyme activities. Dual neutralization of TGF‐β and IL‐6 is notably effective in eliciting splenic and synovial tissue–resident macrophage responses. Synovial macrophage‐derived IL‐10 can elicit protection against septic arthritis via regulating receptor‐activated nuclear factor Kappa‐B ligand (RANKL)/OPG interaction. They also reduced oxidative stress by increasing the activity of antioxidant enzymes including SOD and catalase. Histopathological analysis revealed that dual neutralization of TGF‐β and IL‐6 prevented bone destruction and osteoclastic activity in septic arthritis by promoting the differential functional response of the splenic and synovial macrophages. Additionally, the macrophage‐derived IL‐10 can elicit protection against S. aureus‐induced septic arthritis via regulating RANKL/OPG interaction. Further studies on STAT3 and STAT4 are needed for the understanding of such cross‐talking in resident macrophages of arthritic mice. [ABSTRACT FROM AUTHOR]