Adhesion molecule with Ig-like domain 1 regulates stability of carotid plaque via TGFβ/Smad signaling pathway by interaction with TGFRII.

Reproductive characteristics like ovulation frequency and litter number significantly impact the sheep industry. Transforming growth factor beta 1 ( TGF-β1 ) functions as a multifunctional regulator of key reproductive processes, plays a pivotal role in mammalian ovarian development. However, research on the role of the TGF-β1 gene in the reproduction of Ujimqin and Sonid sheep breeds remains scarce. Thus, this study identified 11 new variants of the TGF-β1 gene in these breeds by direct sequencing. In these 11 variants, the Sonid-linkage disequilibrium (LD)1 and Ujimqin-LD1 (comprising the g.50044287 G > A and g.50044526 G > C polymorphisms) in the 3′ untranslated region of the TGF-β1 gene were significantly associated with litter size in Sonid and Ujimqin ewes ( p < 0.01, p < 0.05, respectively), and the single g.50044837 C > T polymorphism in the 3′UTR was also associated with litter size in Sonid sheep ( p < 0.01) through association analyses. These findings may provide potential genetic markers for improving prolificacy in sheep.

Mucosal delivery (in ovo and oral) of chitosan nanoparticle-based subunit vaccine enhances resistance against Campylobacter jejuni colonization in broiler chickens.

Background Obesity is accompanied by low-grade and chronic pathological development that can worsen pulmonary inflammation and fibrosis. This study investigated the potential of lunasin, a naturally occurring seed peptide with multiple bioactive properties, to attenuate lung inflammation in A549 pulmonary epithelial cells and in C57BL6/J mice fed with the high-fat diet. Methods In vitro , palmitic acid (PA) and lipopolysaccharide (LPS) were used to mimic an obese inflammatory microenvironment. The cultured supernatants were collected for cytokine analysis and cells were collected for specific protein analysis. In vivo , mice were fed a high-fat (HF) diet or an HF diet supplemented with lunasin-enriched soy protein isolated (HFL) from 6 until 22 weeks of age. The lung and spleen samples were collected for future analysis. Results Lunasin inhibited PA- or LPS-induced interleukin (IL)-6, monocyte chemoattractant protein (MCP)-1, and transforming growth factor (TGF)-β secretion. While LPS reduced surfactant protein D (SP-D) expression, lunasin restored SP-D by inhibiting the nuclear factor kappa B (NF-κB) signaling pathway. Additionally, pulmonary fibrosis was induced by TGF-β-induced epithelial-mesenchymal transition (EMT), as indicated by reduced vimentin and preserved E-cadherin expression. However, lunasin did not affect the TGF-β-induced EMT marker in A549 cells. In vivo , HFL-fed mice exhibited lower tumor necrosis factor (TNF)-α and TGF-β levels in lung homogenate compared with HF-fed controls. Lunasin supplementation also enhanced the secretion of T helper cell type 1 (Th1) cytokines, including IL-2 and interferon (IFN)-γ, increased the Th1 (IL-2)/Th2 (IL-4) ratio, and reduced the IL-17A level in splenocytes. Conclusion In summary, in vitro , lunasin attenuated pro-inflammatory cytokines, possibly through enhancing SP-D expression and inhibiting NF-κB signaling in A549 cells. In vivo , dietary lunasin supplementation reduced pulmonary inflammation and modulated splenic cytokine balance. This study reveals for the first time that lunasin is a promising candidate for mitigating obesity-related pulmonary inflammation.

The tumor microenvironment (TME) in breast cancer is shaped by reciprocal interactions between cancer cells and their surrounding stromal populations. Here, we show that breast adipose tissue-derived stromal/stem cells (bASCs) undergo distinct state transitions in response to tumor cues and systemic metabolic status. Using primary bASCs derived from tumor-adjacent and tumor-distant adipose tissues of breast cancer patients with or without obesity, we identify two functionally distinct, tumor-educated stromal phenotypes: a cytokine-rich inflammatory CAF-like (iCAF) state predominating in lean-adjacent bASCs (ln-aT), and a myofibroblastic CAF-like (myCAF) state emerging in obese-adjacent bASCs (ob-aT). Importantly, transforming growth factor β (TGFβ) is sufficient to induce myCAF-like reprogramming in obesity-primed bASCs, while interleukin 1 (IL1)-Janus kinase (JAK) signaling promotes iCAF features. Re-analysis of single-cell RNA-seq data of breast cancer samples reveals an increased TGFβ expression across stromal and immune cell types in individuals with obesity. Mechanistically, IL1 receptor blockade (anakinra) or JAK inhibition (AZD1480) reverses both iCAF and myCAF phenotypes and functionally suppresses stromal-driven epithelial-mesenchymal transition as well as cancer stemness in breast cancer cells. These findings establish a mechanistic link between obese cues, stromal plasticity, and breast cancer progression, and reveal IL1/JAK signaling as a tractable axis to therapeutically reprogram the breast cancer stroma.

Chronic liver injury is accompanied by coordinated disturbances in lipid trafficking and inflammatory-fibrogenic signaling. Transforming growth factor beta 1 (TGF-β1) signaling has been implicated in hepatic fibrogenesis and tumor-associated remodeling and may co-vary with disturbances in lipid trafficking. Lisosan G (LG), a fermented wheat-derived nutraceutical, has reported antioxidant and anti-inflammatory activity and may influence these interconnected pathways. This study evaluated whether dietary LG alters the lipid composition of plasma lipoprotein fractions and hepatic TGF-β1 levels across distinct liver contexts. Seventy-two female Wistar rats were randomized into nine groups (n = 8/group) defined by liver condition, consisting of healthy control (Control), non-neoplastic liver (PH), and neoplastic liver injury (HCC; PH followed by diethylnitrosamine, DEN), and diet (standard diet, SD + 2.5% LG, or SD + 5% LG). Plasma lipoproteins (VLDL, LDL, HDL1, HDL2) were isolated by stepwise KBr density-gradient ultracentrifugation, and cholesterol (TC), phospholipids (PL), and triacylglycerols (TG) were quantified in each fraction. Hepatic TGF-β1 was measured by ELISA and normalized to total protein. LG effects depended strongly on baseline liver status, with significant Condition × Diet interactions for most lipid endpoints and for hepatic TGF-β1. In healthy rats, LG produced fraction-selective remodeling rather than uniform lipid lowering, including increased VLDL-TG at both doses and non-linear changes in cholesterol distribution across LDL and HDL subfractions. After PH, LG broadened lipid remodeling, including reduced VLDL-PL, increased VLDL-TG (both doses), and an increase in LDL-TC at 5% LG, accompanied by marked changes in HDL1/HDL2 cholesterol partitioning. In HCC, LG induced pronounced, often dose-dependent increases in LDL-associated lipids (LDL-PL, LDL-TG, LDL-TC) and increased HDL1-TC while decreasing HDL2-TC. Hepatic TGF-β1 was elevated in PH and further increased in HCC versus controls; LG reduced hepatic TGF-β1 in a condition-dependent manner, with the strongest reduction at 5% LG in HCC. Dietary Lisosan G remodels circulating lipoprotein lipid composition in a liver-status-dependent manner and is associated with reduced hepatic TGF-β1 abundance in injured liver, most prominently in neoplastic injury. These findings are consistent with the notion that nutraceutical interventions may show stronger phenotypic effects under perturbed metabolic-fibrogenic states than under stable physiology, while highlighting the need for mechanistic work to distinguish altered lipoprotein secretion from changes in peripheral clearance and to assess pathway-level TGF-β signaling.

Matrix metalloproteinases (MMPs) and their endogenous inhibitors, tissue inhibitors of metalloproteinases, regulate extracellular matrix remodeling and neural tissue homeostasis. Transforming growth factor β1 modulates inflammation, ECM deposition, and tissue repair. Dysregulation of these pathways may contribute to congenital spinal dysraphism. This study quantified cerebrospinal fluid MMP-9, TIMP-1, and TGF-β1 in neonates with myelomeningocele and evaluated their potential as pathogenic mediators or biomarkers. Preoperative CSF samples were collected from 48 neonates with isolated myelomeningocele (Group 1) and 24 age-matched controls with sterile CSF (Group 2). MMP-9, TIMP-1, and TGF-β1 concentrations were measured via ELISA. Intergroup differences were analyzed, and Pearson correlation assessed relationships among biomarkers. CSF MMP-9 levels were significantly elevated in neonates with myelomeningocele compared to controls (57.12 ± 64.19ng/mL vs. 11.29 ± 6.79ng/mL; p < 0.05). TIMP-1 (368.78 ± 220.08ng/mL vs. 450.04 ± 390.22ng/mL) and TGF-β1 (2.32 ± 1.44ng/mL vs. 2.524 ± 1.07ng/mL) did not differ significantly. MMP-9 correlated positively with TIMP-1 across all subjects (p < 0.05) and with TGF-β1 within the myelomeningocele group (p < 0.05), indicating coordinated regulation of ECM degradation and fibrogenic signaling. Elevated CSF MMP-9 in neonates with myelomeningocele suggests dysregulated ECM remodeling may be associated with neural tube malformation. MMP-9 may serve as a biomarker of ECM dysregulation, providing insights into the pathogenesis of congenital spinal dysraphism and identifying possible targets for future therapeutic interventions.

Investigating the effect of alpha-lipoic acid and retinoic acid eye drops on re-epithelialization, inflammation, and angiogenesis in a rabbit model of corneal wound healing through ICAM-1 and MMP-9 pathways.

Exosomes derived from ovarian cancer promote the progression of ovarian cancer through macrophage M2 polarization mediated by the THBS1/TGFBI signaling axis.

Advances in medicine lead to an increase in the number of patients with concurrent heart disease and cancer. We investigated the effect of cardiac surgery under cardiopulmonary bypass (CPB) on the blood levels of growth factor cytokines, and assessed whether it might affect the proliferation of pancreatic cancer cells. Ten consenting patients undergoing elective cardiac surgery with the use of CPB under general anesthesia. We obtained patients' serum at each of the following time points: between median sternotomy and before activating CPB (pre-CPB), and after activating CPB (post-CPB). Growth factor cytokines in plasma samples were analyzed using cytokine array. Proliferation assay was performed using pancreatic cancer cells (PANC-1 and PK-1) incubated in culture medium containing patients' serum at a concentration of 30%. The levels of cytokines and proliferation of cancer cells were compared between pre-CPB and post-CPB samples. Plasma and serum from eight patients were analyzed. PANC-1 and PK-1 cells proliferated significantly more following incubation with post-CPB (125%, P = 0.014 and 120%, P = 0.030, respectively) than pre-CPB samples. The following cytokines were significantly increased post-CPB compared to pre-CPB: amphiregulin (42 fold), basic fibroblast growth factor (5 fold), heparin binding-epidermal growth factor (9199 fold), hepatocyte growth factor (9 fold), insulin-like growth factor-binding protein (IGFBP)-1 (11 fold), IGFBP-4 (15 fold), insulin-like growth factor-2 (2 fold), macrophage colony-stimulating factor (2 fold), neurotrophin-4 (10 fold), transforming growth factor beta-3 (2 fold), vascular endothelial growth factor (VEGF)-A (3 fold), and VEGF receptor 2 (3 fold). These results suggest that cardiac surgery with CPB increases the plasma levels of several growth factors, thus potentially promoting pancreatic cancer cell proliferation, as seen in the in vitro study using serum from non-cancer patients. registration number UMIN000053046, date of registration December 12th, 2023.

Transforming growth factor beta (TGF-β) is a pleiotropic cytokine and participates in multiple cellular processes, such as cell development, proliferation, epithelialmesenchymaltransition (EMT), and immune responses through SMAD-dependent or SMAD-independent signaling pathways. Notably, TGF-β signaling plays a dual role in tumors, acting as a potent tumor suppressor during early tumorigenesis by inducing apoptosis or cell-cycle arrest while promoting tumor transformation, progression and metastasis in advanced stage through multidimensional mechanisms. Moreover, it is abundant and functions as a master immune checkpoint in the tumor microenvironment (TME), fostering the development of numerous targeted therapies to rectify its aberrant activity in tumors in the past decades. Thus, a comprehensive overview of the pathologic roles, molecular mechanisms and therapeutic potentials of TGF-β signaling in tumors will benefit both the basic and clinical cancer research. Here, we review the complex biology and context-dependent functions of the TGF-β superfamily in regard to tumor, highlighting how it regulates the latter's development, growth, and dissemination by mainly targeting tumor cells, tumor-associated fibroblasts and various immune cells. We also summarize recent advances in the preclinical and clinical development of different types of TGF‑β‑targeting agents, and discuss their therapeutic potentials and challenges as well as approaches to improve the safety and efficacy of TGF-β pathway-targeted therapy in cancers. Through the summary of known knowledge and the latest updates, this review may provide a general picture on the biological functions of TGF-β in tumors, and facilitate the clinical implications of TGF-β-targeted therapy in tumor patients.

Pirfenidone as a Pleiotropic Antifibrotic Agent in Metabolic Steatohepatitis: From Mechanisms to Clinical Evidence.

Bioactive compounds and multitarget action mechanism of Erzhi pills in alleviating metabolic dysfunction-associated steatohepatitis.

A genome-wide association study (GWAS) identified neuron navigator 3 (NAV3) as a potential genetic determinant of myocardial recovery in dilated cardiomyopathy (DCM). This study aimed to understand its functional role in cardiac pathophysiology by leveraging omics approaches. Single-cell RNA-seq transcriptomic data from previously published adult human hearts indicate that NAV3 expression is highest in cardiac fibroblasts, suggesting its functional role in these cells. In vitro, stimulation of primary human ventricular cardiac fibroblasts with transforming growth factor β1 (TGF-β1) induced NAV3 expression in a dose and time-dependent manner. Small-interfering-RNA-mediated knockdown of NAV3 significantly attenuated TGF-β1-induced fibroblast activation, reducing the expression of α-smooth muscle actin (α-SMA), collagens, and fibronectin. RNA sequencing of NAV3-silenced fibroblasts, confirmed by Western blot, revealed upregulation of cell cycle regulators and downregulation of profibrotic markers, suggesting that NAV3 facilitates TGF-β1-induced cell cycle arrest and fibroblast-to-myofibroblast transition. Notably, NAV3 silencing did not alter canonical SMAD2/3 phosphorylation, implying a role for NAV3 in modulating fibrotic signaling through other pathways. Our findings provide functional and mechanistic insights into NAV3's novel role in cardiac fibrosis, showing that reduced NAV3 expression attenuates TGF-β1-mediated fibroblast activation by regulating cell cycle signaling. These results support further investigation of NAV3 as a potential modulator of cardiac fibrosis and myocardial recovery in DCM.NEW & NOTEWORTHY This study uncovers a previously unrecognized role for NAV3 in TGF-β1-driven cardiac fibroblast activation. We show that NAV3 facilitates profibrotic remodeling through noncanonical signaling and cell cycle arrest, independently of SMAD2/3. These findings position NAV3 as a novel regulator of fibroblast phenotype and a potential modulator of cardiac fibrosis.

Cancer-associated fibroblasts (CAFs) transdifferentiated from hepatic stellate cells (HSCs) are a critical determinant of liver metastasis of colorectal cancer (CRC). However, the mechanisms behind transforming growth factor β (TGF-β)-stimulated activation of HSCs into CAFs remain poorly understood. Immunoprecipitation coupled with mass spectrometry identified tuftelin 1 (TUFT1) as a novel TGF-β receptor II (TβRII) binding protein in primary human HSCs and immortalized LX2 cells. TUFT1 interacts with TβRII via its fragments (amino acids 1-86, 87-157), protecting TβRII from lysosomal degradation to facilitate TGF-β signaling and myofibroblastic activation of HSCs. Mechanistically, TUFT1 competes with caveolin-1 for TβRII binding, retrieving TβRII from the lipid rafts/caveolae-mediated degradation pathway and sorting it into the endosome-mediated trafficking and signaling pathway. Clinically, TUFT1 expression was confirmed in the CAFs of patient-derived colorectal cancer liver metastasis (CRCLM) tissues. Both protein and transcript analyses revealed higher TUFT1 expression in the CAFs of CRCLM than in HSCs. Furthermore, bulk RNA sequencing indicated that knocking down TUFT1 altered the TGF-β transcriptome of HSCs and suppressed HSC expression of tumor-promoting factors. In HSC/CRC co-implantation and portal vein tumor injection mouse models, targeting TUFT1 of HSCs inhibited HSC activation and restricted CRC growth in both subcutaneous and hepatic sites. Taken together, our findings uncover the novel function of TUFT1 in the hepatic tumor microenvironment, highlighting its role as a critical regulator of HSC activation and the pro-metastatic hepatic niche via promoting TβRII protein stability. Targeting TUFT1 in HSCs presents a promising therapeutic approach for combating CRCLM.

Fibrosis is a hallmark of organ failure observed after chronic epithelial injury and inflammation. The transforming growth factor beta (TGF-β) is the master regulator of fibrogenesis, so blockade of the TGF-β pathway is a potential treatment strategy for fibrosis; however, the therapeutic potential of pan-TGF-β blockade is limited by side effects. We generated SOF10, a humanized antibody that targets latent TGF-β1 and selectively blocks protease- and integrin αvβ8-mediated latent TGF-β1 activation. We conducted gene expression and histological analyses in nonalcoholic steatohepatitis (NASH)/liver fibrosis and renal interstitial fibrosis models. We also evaluated the combination effect of SOF10 with an immune checkpoint inhibitor in a syngeneic mouse model and performed safety studies in mice and monkeys. Here we show that SOF10 reduces fibrosis in NASH/liver fibrosis and renal interstitial fibrosis models and improves renal function in a chronic kidney disease model. Furthermore, the combination of SOF10 with an anti-PD-L1 antibody decreases tumor growth in a syngeneic mouse model. SOF10 demonstrates safety in both mice and monkeys. Selective blockade of latent TGF-β1 activation represents a promising approach for treating a broad range of fibrotic diseases and cancers. By specifically targeting TGF-β1, SOF10 may offer a safer and more effective therapeutic option compared to non-selective TGF-β inhibitors. This strategy has the potential to transform the treatment paradigm for fibrosis-related conditions.

BACKGROUNDProprotein convertase subtilisin/kexin type 9 (PCSK9) is abundantly expressed by hepatocytes and regulates the uptake of low-density lipoprotein by these parenchymal cells. Little research has been conducted on PCSK9 expression in non-hepatocyte liver cells.AIMTo investigate PCSK9 levels in the supernatant of different liver cells and its regulation in hepatic stellate cells (HSCs).METHODSPCSK9 levels were measured in the cell culture medium of primary human hepatocytes, HepG2 and Huh7 cells, primary human HSCs, the HSC cell line LX-2, primary human Kupffer cells, and primary human sinusoidal endothelial cells. The effects of cytokines, adipokines, lipopolysaccharide, transforming growth factor beta (TGF-β) and ligands of nuclear receptors on PCSK9 levels in LX-2 cells during 24 hours of culture were determined using enzyme-linked immunosorbent assay.RESULTSPrimary human hepatocytes, HepG2, Huh7, HSCs, and LX-2 cells secreted significant levels of PCSK9. There were low levels of PCSK9 in the supernatant of Kupffer cells and sinusoidal endothelial cells. Interleukin-6 reduced PCSK9 in LX-2 cells to 86% of controls and lipopolysaccharide increased it by 7%, whereas tumor necrosis factor, as well as exogenous adiponectin and leptin had no effect. Chemerin-156, but not chemerin-155 or chemerin-157 isoform overexpressed in LX-2 cells, reduced PCSK9 to 84% of the controls. TGF-β reduced PCSK9 in LX-2 cell culture media to 68% of controls and lowered its cellular level. Activation of liver X receptor but not farnesoid X receptor or peroxisome proliferator-activated receptor gamma, reduced PCSK9 levels by 42% in LX-2 cell culture medium.CONCLUSIONProfibrotic TGF-β and the antifibrotic liver X receptor ligand both reduced PCSK9 in LX-2 medium, showing that PCSK9 is not a marker of HSC activation.

BACKGROUNDThe incidence of diabetic atherosclerosis (DMA) is increasing worldwide, but its pathogenesis remains incompletely understood. In addition to cardiovascular complications, bladder dysfunction is one of the common comorbidities associated with DMA but is often refractory to current treatments.AIMTo investigate the therapeutic effect of human amniotic fluid stem cell-derived extracellular vesicles (hAFSC-EVs) on the recovery of bladder dysfunction in DMA rats.METHODSEighty rats were divided into normal control, streptozotocin-induced diabetic rats, diabetic rats subjected to arterial balloon endothelial injury of common iliac artery (DMA), and DMA rats treated with hAFSC-EVs (DMA + hAFSC-EVs). At 4 weeks and 12 weeks after DMA induction, levels of blood glucose, total cholesterol, triglyceride, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, homeostasis model assessment (HOMA)-insulin resistance, and HOMA-β were measured. Cystometry, common iliac artery wall thickness, and bladder tumor necrosis factor (TNF)-α, interleukin (IL)-6, transforming growth factor (TGF)-β1, Smad3, connective tissue growth factor (CTGF) and fibronectin were also evaluated.RESULTSBladder weight and blood glucose, triglyceride, HOMA-insulin resistance, common iliac artery intima thickness, voided volume, intercontraction interval, bladder capacity, and mRNA expression of TNF-α, IL-6, TGF-β1, Smad3, CTGF and fibronectin were significantly increased at 4 weeks and 12 weeks after induction, while the HOMA-β level decreased at 4 weeks and 12 weeks, and the high-density lipoprotein cholesterol level decreased at 12 weeks. hAFSC-EVs treatment in DMA rats significantly reduced bladder weight and blood glucose, thickness of common iliac arterial intima, voided volume, intercontraction interval and bladder capacity at 4 weeks. The mRNA expression of TNF-α, TGF-β1, and CTGF in DMA rats treated with hAFSC-EVs were significantly decreased at 4 weeks, while the mRNA expressions of IL-6 and Smad3 were significantly decreased 12 weeks.CONCLUSIONhAFSC-EVs treatment can help restore DMA-induced bladder dysfunction, which is associated with lowered blood glucose levels, reduced arterial wall thickness, and decreased TNF-α, IL-6, TGF-β1, Smad3, and CTGF expression.

Beyond their canonical role in promoting G1/S progression, the complexes formed by cyclin D and cyclin-dependent kinase (CDK) 4/6 have emerged as contributors to enhanced cell migration. However, a direct link between this complex and cytoskeletal remodeling during cell motility has remained poorly understood. Here, we show that CDK4/6 inhibition in HeLa cells disrupts lamellipodia formation and subsequent focal adhesion assembly, leading to a reduction in cell migration and invasion. Notably, CDK4, but not CDK6, in complex with cyclin D1/D2, localizes to membrane ruffles to facilitate cytoskeletal reorganization. Mechanistically, proteomic and phosphoproteomic analyses revealed that CDK4 inhibition attenuates the transforming growth factor β (TGFβ) pathway via reduced Smad3 phosphorylation at Thr8, downregulating integrin subunits (α5, α6, and β1). Furthermore, CDK4 inhibition significantly decreased focal adhesion kinase (FAK) phosphorylation at Tyr397 and Rac1-GTP levels. Importantly, the resulting migration defect was largely restored by activation of either Rac1 or FAK. Thus, our data support a model in which cyclin D1/D2-CDK4 promotes phosphorylation of Smad3, leading to upregulation of integrin subunits, activation of FAK and Rac1, and consequent lamellipodia formation and cell migration. These findings provide direct evidence that CDK4 regulates actin cytoskeletal reorganization during cell migration and suggest that CDK4/6 inhibitors may dampen cytoskeleton-dependent tumor invasion, in addition to their antiproliferative effects.

Extracellular Vesicle-Mediated Communication Between Anterior Cruciate Ligament and Bone Marrow Cells Modulates Hamstring Tenocyte Behavior and Apoptosis.