Β1 Expression Research Articles

Integrin β2 Plays a Significant Role in Therapeutic Angiogenesis Through Hematopoietic Stem Cell Transplantation

The efficacy of hematopoietic stem cell (HSC) therapy for cerebral infarction has been previously demonstrated. However, the lack of response in some patients has hindered its widespread use. To establish HSC therapy as a standard treatment, it is important to examine the causes of non-responsiveness. In this study, we aimed to identify the specifications of transplanted cells based on their therapeutic mechanisms to predict treatment success. We found that HSC therapy activates injured cerebral endothelial cells via gap junctions because cell adhesion between HSCs and the endothelium plays an essential role in cellular communication via gap junctions. The expression of the adhesion molecule integrin β2 (CD18) in CD34-positive (CD34+) cells was identified as critical for the therapeutic effect on cerebral infarction in a murine model. Cells with low CD18 expression exhibited a weaker therapeutic effect than cells with high CD18 expression, even when the same number of HSCs was administered. The expression of CD18 in CD34+ cells can be used as a specification marker for transplanted HSCs and is useful for identifying non-responders. Furthermore, quantification of CD18 expression is crucial for evaluating the cellular potential of cell-based therapies for diseases where therapeutic effects are mediated through cell adhesion.

TPMS-Gyroid Scaffold-Mediated Up-Regulation of ITGB1 for Enhanced Cell Adhesion and Immune-Modulatory Osteogenesis.

The porous structure is crucial in bone tissue engineering for promoting osseointegration. Among various structures, triply periodic minimal surfaces (TPMS) -Gyroid has been extensively studied due to its superior mechanical and biological properties. However, previous studies have given limited attention to the impact of unit cell size on the biological performance of scaffolds. In this research, four TPMS-Gyroid titanium scaffolds with different unit cell sizes (TG15, TG20, TG25, and TG30) are fabricated using Selective Laser Melting (SLM) to explore their effects on osseointegration. Mechanical tests revealed that TG15 and TG20 exhibited superior compressive strength. In vitro experiments demonstrated that TG20 facilitated better cell adhesion through robust integrin protein expression initially, which subsequently enhanced cell proliferation and osteogenic differentiation. Furthermore, macrophages on TG20 showed higher Integrin β1 (ITGB1) expression, promoting their polarization to the M2 phenotype, which suppressed inflammation, fostered bone integration, and angiogenesis. In vivo studies confirmed TG20's effectiveness in promoting bone ingrowth by reducing inflammation. This study highlights TG20's structural advantages, making it a promising bone scaffold with exceptional osteogenic and angiogenic properties through osteoimmune microenvironment modulation. Therefore, TG20 holds significant potential for applications in bone tissue engineering.

Structural elucidation and anti-asthmatic effects of semi-crystalline polysaccharides from Lignosus rhinocerotis (Cooke) Ryvarden.

Asthma is a chronic respiratory disease characterized by airway inflammation. Lignosus rhinocerotis (LR), a medicinal mushroom rich in polysaccharide, has been traditionally used to treat various diseases, including asthma. This study aimed to fractionate, characterize and evaluate the anti-asthmatic effects of polysaccharides from LR (LRP). LRP was isolated and characterized using high-performance liquid chromatography (HPLC), x-ray diffraction analysis (XRD), fourier transform infrared spectrometry (FTIR) and nuclear magnetic resonance (NMR) spectroscopy. In an OVA-induced asthma model, BALB/c mice were sensitized, challenged, and intranasally treated with LRP. Experimental findings demonstrated that LRP exhibits a semi-crystalline nature with predicted structure of → 4)-α-D-Glcp-(1 → and → 3)-β-D-Glcp-(1→. LRP significantly reduced eosinophilia, Th2 cytokines, and IgE levels. Histological analysis revealed LRP's ability to decrease epithelial damage and epithelial and smooth muscle thickness in lung. Reductions in inflammatory cell infiltration, mucus production, and transforming growth factor (TGF)-β1 expression were observed, although not statistically significant. Gene expression analysis indicated that LRP significantly downregulated the inducible nitric oxide synthase (iNOS) expression. This study highlights a detailed structural analysis of LRP and its potential as an alternative for the management of asthma. Further research is needed to elucidate the precise mechanisms of action and optimize its therapeutic application.

Exploring the Active Constituents of Andrographis paniculata in Protecting the Skin Barrier and the Synergistic Effects with Collagen XVII.

Andrographis paniculata is mainly used to treat skin inflammations, wounds, and infections. In this study, Andrographis Herba, the aerial part of the plant, was proven to increase the viability of UVB-damaged HaCat cells and reduce reactive oxygen species levels. The chemical composition of Andrographis Herba extract (AHE) was analyzed using UPLC-Q-TOF-MS, and diterpene lactones were identified as its primary constituents. Then, the fraction of diterpene lactones was prepared and exhibited similar effects to AHE. AHE, its diterpene lactones component, and its representative constituent andrographolide all decreased the expression of IL-1β, IL-6, and CDKN1A. Furthermore, the protective effects of AHE and its active ingredients on UVB-damaged epidermal stem cells were investigated. Notably, the combined treatment with andrographolide and collagen XVII enhanced the viability of UVB-damaged epidermal stem cells, increased the expression of stemness markers integrin β1 and p63, and decreased the expression of the differentiation marker keratin 10. This combination demonstrated significant synergy in maintaining skin homeostasis, which provides evidences for the development of skin-protective products.

High matrix stiffness accelerates migration of hepatocellular carcinoma cells through the integrin β1-Plectin-F-actin axis

BackgroundAbundant research indicates that increased extracellular matrix (ECM) stiffness significantly enhances the malignant characteristics of hepatocellular carcinoma (HCC) cells. Plectin, an essential cytoskeletal linker protein, has recently emerged as a promoter of cancer progression, particularly in the context of cancer cell invasion and metastasis. However, the responsiveness of plectin to changes in ECM stiffness and its impact on HCC progression remain unclear. In this study, we aimed to investigate whether plectin responds to variations in ECM stiffness and to explore its involved molecular mechanisms in regulating HCC cell migration.ResultsOur results showed that, when compared with control group (7 kPa), high ECM stiffness (53 kPa) boosts HCC cell migration by upregulating plectin and integrin β1 expression and increasing F-actin polymerization. Knockdown of integrin β1 negated the high stiffness-upregulated plectin expression. Furthermore, reducing either plectin or integrin β1 levels, or using latrunculin A, effectively prevented the high ECM stiffness-induced F-actin polymerization and HCC cell migration.ConclusionsThese findings demonstrate that integrin β1-plectin-F-actin axis is necessary for high matrix stiffness-driven migration of HCC cells, and provide evidence for the critical role of plectin in mechanotransduction in HCC cells.

Gingival keratinocyte adhesion on atomic layer-deposited hydroxyapatite coated titanium.

This study aimed to evaluate the effects of the atomic layer deposited hydroxyapatite (ALD-HA) coating of the titanium (Ti) surface on human gingival keratinocyte (HGK) cell adhesion, spreading, viability, and hemidesmosome (HD) formation. Grade 2 square-shaped Ti substrates were used (n = 62). Half of the substrates were ALD-HA coated, while the other half were used as non-coated controls (NC). The ALD-HA surface was characterized with scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS) analysis. The initial cell adhesion and HD formation of HGKs were evaluated after a 24-h cultivation period. The cell proliferation was assessed by cultivating cells for 1, 3, and 7d. The expression levels of the integrin mediating cell adhesion were detected with the Western Blot method. In addition, cell spreading and expression of the proteins mediating cell adhesion were imaged using a confocal microscope. SEM-EDS analysis demonstrated the formation of HA on the ALD-HA surfaces. The relative cell attachment was significantly higher (p < .05) on the ALD-HA compared to the NC surface after 1 and 3 d of cell culture. No significant difference was found in integrin α6 or β4 expression. The microscope evaluation showed significantly increased cell spreading with peripheral HD expression on ALD-HA compared to the NC surfaces (p = .0001). Moreover, laminin γ2 expression was significantly higher on the ALD-HA than on the NC surfaces (p < .001). Compared to the NC Ti surface, the ALD-HA coating has favorable effects on HGK proliferation, growth, and cell spreading. This indicates that the ALD-HA coating has good potential for improving mucosal attachment on implant surfaces.

Regeneration of the skin wound by two different crosslinkers: In vitro and in vivo studies.

For designing a suitable hydrogel, two crosslinked Alginate/ Carboxymethyl cellulose (Alg/CMC) hydrogel, using calcium chloride (Ca2+) and glutaraldehyde (GA) as crosslinking agents were synthesized and compared. All samples were characterized by Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM), Blood compatibility (BC), Blood clotting index (BCI), weight loss (WL), water absorption (WA), pH, and Electrochemical Impedance Spectroscopy (EIS). Cell viability and cell migration were investigated using the MTT assay and the wound scratch test, respectively. Besides, the wound healing potential of prepared hydrogels was evaluated on the rat models with full-thickness skin excision. To further investigation, TGF β1, IGF-I, COL1, ACT-A (alfa-SMA), and GAPDH expression levels were also reported by RT-PCR. Water absorption and weight loss properties were compared between different crosslinker agents, and the most nontoxic crosslinker concentration was determined. We have shown that GA (20 µl/ml) and Ca2+ (50 or 75 mM) enhanced the physical stability of Alg-CMC hydrogel, and they are nontoxic and suitable crosslinkers for wound dressing applications. Although in vivo assessments indicated that the GA (20 µl/ml) had a cytotoxic effect on tissue repair, Ca2+ (75 mM) boosted the wound healing process. Further, RT-PCR results revealed that TGF β1, IGF-I, COL1, ACT-A (alfa-SMA), and GAPDH expression levels were increased in GA (20 µl/ml). Moreover, this trend is the opposite in the Ca2+ (75 mM) treatment groups. This research shows that Ca2+ (75 mM) boosts tissue regeneration and wound healing process.

Donkey milk supplementation alleviates renal fibrosis of chronic kidney disease by enhancing anti-inflammatory ability.

Donkey milk, rich in essential nutrients, is not only able to meet the nutritional needs of patients with chronic kidney disease (CKD) according to the National Kidney Foundation's Kidney Disease Outcomes Quality Initiative Clinical Practice Guideline for Nutrition in CKD, but also has strong anti-inflammatory properties. Inflammation is the main driving force of renal fibrosis, which directly determines renal function in CKD. However, the impact of donkey milk on CKD and the underlying mechanisms remain unclear. We hypothesized that donkey milk supplementation could enhance anti-inflammatory abilities in individuals with CKD, thereby inhibiting renal fibrosis and improving renal function. Therefore, we investigated the influence of a 6-wk treatment with donkey milk on unilateral ureteral ligation-induced CKD rats. The results demonstrated that the level of miR-223-3p was decreased in the kidneys of CKD rats compared with normal rats. However, donkey milk supplementation activated miR-223-3p, thereby inhibiting the expression of inhibitor of kappa-B kinase (IKKα) and nuclear factor kappa-B (NF-κB) proteins. Consequently, the expression of inflammatory factors such as monocyte chemotactic protein-1 (MCP-1) and IL-6 was suppressed, leading to reduced aggregations of macrophages in the kidney and decreased transforming growth factor β1 (TGF-β1) expression. Ultimately, renal fibrosis was inhibited, which was confirmed by decreased collagen type I α 1 (COL1A1) mRNA, thereby the renal function of CKD rats was improved. These results indicate that miR-223-3p may be another potential therapeutic target for CKD. Moreover, the contribution of donkey milk to alleviating the inflammatory response and renal fibrosis of CKD is obvious, indicating that donkey milk may be a new functional food for CKD patients to promote health.

A novel solvent-extruded tacrolimus-eluting suture for attenuated inflammation and scarring in skin repair

This study aimed to design an absorbable suture that locally delivers the immunosuppressive agent, tacrolimus (FK506), to attenuate scar tissue formation by modulating inflammation. As the suture degrades, it releases FK506 directly into the repair site. The poly (l-lactide-co-caprolactone) (PLLA-PCL) polymer and 0.2 % wt FK506 were co-extruded using a solvent extrusion technique to generate the drug-eluting sutures. The properties of the generated sutures were assessed regarding FK506 release in vitro, wound closure in an in vivo murine model, murine skin histology for structural and inflammatory markers, and mechanical strength of both sutures and healed murine skin. The generated suture consistently released FK506 at ≤10 ng/day rates. Following annealing, the sutures retained 60 % of their anticipated tensile strength, exhibiting enhanced ductility. The histological examination and in vivo murine study indicated that wounds treated with FK506-eluting PLLA-PCL sutures produced a thinner epidermal layer, enhanced integrin β4 expression, increased keratinocyte presence, attenuated inflammation, and reduced scarring, demonstrating healed tissue improvements compared with those treated with the placebo, poly (glycolic-co-caprolactone) (PGCL), sutures. Mechanical evaluations of healed tissue indicated that drug-PLLA-PCL sutures did not reduce the mechanical strength of the skin compared to placebo PGCL sutures. The FK506-eluting PLLA-PCL sutures show promising results in advancing wound healing by offering targeted drug delivery. This innovative method can improve wound healing and provide a streamlined approach to scar management. Continued research, optimization, and broad-scale studies are needed to refine the manufacturing process and ensure that the properties of the suture meet clinical requirements.

Innovative 3D-Printed Titanium Specimens Favor a Modulation of Inflammation in Dental Pulp Stem Cells During Liposome-Triggered Mineralization.

developing customized titanium specimens, with innovative surfaces, is a suitable strategy to overcome implant failure. Additionally, a faster and efficient osteogenic commitment assists tissue regeneration. To investigate the interplay between inflammation and differentiation upon implantation, Dental Pulp Stem Cells (DPSCs) were cultured on 3D-printed titanium owning an internal open cell form, administering osteogenic factors by a liposomal formulation (LipoMix) compared to traditional delivery of differentiation medium (DM). osteogenic differentiation was evaluated by western blot, by measuring β1 integrin expression, and by and real-time RT-PCR, by measuring SP7 and Collagen I gene expression; while.angiogenesis was characterized by measuring VEGF secretion levels. Matrix mineralization was assessed by means of Alizarin Red Staining, cell adhesion and inflammation responses through western blot, enzymatic and ELISA assays evaluating Nrf2 expression, catalase activity and Prostaglandin E2 secretion, respectively. LipoMix enhances cell proliferation and adhesion, as revealed by increased integrin β1 expression. Mineralized matrix deposition, SP7 gene expression, Collagen I release and Alkaline Phosphatase activity appear increased in LipoMix condition. Additionally, the redox-sensitive transcription factor Nrf2 is overexpressed at the earliest experimental times, triggering the catalase activity. data reported confirm that internal topography and post-production treatments on titanium surfaces dynamically and positively condition the DPSC progress towards the osteogenic phenotype, moreover, the combination with LipoMix fastens the positive modulation of inflammation under osteogenic conditions. Therefore, the development of customized surfaces along with the administration of differentiating factors enclosed in a liposomal delivery system, could represent a promising and innovative tool in regenerative dentistry.

Expression of interleukin-8 and integrin β3 predicts prognosis of patients with hepatocellular carcinoma after hepatectomy.

As important components in the tumor microenvironment, interleukin-8 (IL-8) and integrin β3 play a key role in the progression and metastasis of hepatocellular carcinoma (HCC). This study aimed to determine the expression of IL-8 and integrin β3 and their prognostic value in patients with HCC after hepatectomy. We investigated the expression of IL-8 and integrin β3, their clinical significance, as well as their correlation in the cancer tissue of 130 patients with HCC using immunohistochemistry. The prognostic value of IL-8 and integrin β3 was investigated through the follow-up of patients with HCC after hepatectomy. In HCC, IL-8 expression had a positive correlation with integrin β3 expression. Increased expressions of IL-8 and integrin β3 were indicators of tumor progression and poor prognosis in patients with HCC after hepatectomy. IL-8 positive specimens exhibited a higher proportion of macrovascular invasion, larger tumor size, poor differentiation, and advanced tumor-node-metastasis (TNM) stage (P < .05, respectively). Integrin β3 positive group exhibited a higher proportion of TNM III-staged tumors (P < .05). The results indicated that macrovascular invasion, advanced TNM stage, and integrin β3 expression were independent unfavorable prognostic factors in HCC after hepatectomy. Integrin β3 expression was proved to be an independent unfavorable prognostic factor in HCC after hepatectomy. Targeting integrin β3 might be a potential therapeutic approach in preventing tumor progression in HCC.

YKL40/Integrin β4 Axis Induced by the Interaction between Cancer Cells and Tumor-Associated Macrophages Is Involved in the Progression of High-Grade Serous Ovarian Carcinoma.

Macrophages in the tumor microenvironment, termed tumor-associated macrophages (TAMs), promote the progression of various cancer types. However, many mechanisms related to tumor-stromal interactions in epithelial ovarian cancer (EOC) progression remain unclear. High-grade serous ovarian carcinoma (HGSOC) is the most malignant EOC subtype. Herein, immunohistochemistry was performed on 65 HGSOC tissue samples, revealing that patients with a higher infiltration of CD68+, CD163+, and CD204+ macrophages had a poorer prognosis. We subsequently established an indirect co-culture system between macrophages and EOC cells, including HGSOC cells. The co-cultured macrophages showed increased expression of the TAM markers CD163 and CD204, and the co-cultured EOC cells exhibited enhanced proliferation, migration, and invasion. Cytokine array analysis revealed higher YKL40 secretion in the indirect co-culture system. The addition of YKL40 increased proliferation, migration, and invasion via extracellular signal-regulated kinase (Erk) signaling in EOC cells. The knockdown of integrin β4, one of the YKL40 receptors, suppressed YKL40-induced proliferation, migration, and invasion, as well as Erk phosphorylation in some EOC cells. Database analysis showed that high-level expression of YKL40 and integrin β4 correlated with a poor prognosis in patients with serous ovarian carcinoma. Therefore, the YKL40/integrin β4 axis may play a role in ovarian cancer progression.

Elevated MMP-9, Survivin, TGB1 and Downregulated Tissue Inhibitor of TIMP-1, Caspase-3 Activities are Independent of the Low Levels miR-183 in Endometriosis.

This study aimed to measure the correlation between miR-183 and gene expression that regulates apoptosis and adhesion mechanism that may be linked to the pathogenesis of endometriosis. Forty-four subjects, including 22 control subjects, participated in this study. We collected ectopic endometriosis and endometrial samples. For the control, the sample was taken from endometrial tissue through pipelle biopsy. RNA was extracted from all tissues using RNA mini kit, and the expression was assessed using quantitative-real time PCR. Relative mRNA and miRNA expression were presented using the formula of the Livak method. The data were statistically analyzed using GraphPad Prism 8. The expression of Caspase-3, Survivin, Integrin β1 (ITGB1), matrix metalloproteinase-9 (MMP-9), and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) (adhesion- and apoptosis-related gene) were calculated using the relative expression method. We found significant differences in Caspase-3, Survivin, ITGB1, MMP-9, and TIMP-1 expression between ectopic endometriosis tissues of women with endometriosis compared to healthy endometrium. MMP-9, Survivin, and ITGB1 was significantly increased in the endometriosis group, while Caspase-3, TIMP-1, and miR-183 were significantly reduced in the endometriosis group. No correlation was found between the expression level of miR-183 and Caspase3, Survivin, ITGB1, and Cadherin in both tissue types. Despite the difference in expression levels of miR-183 and associated adhesion- and apoptosis-related genes, there was no significant association between miR-183 with specific adhesion and apoptosis genes in endometriosis tissue.

Short-Term Exposure to Foodborne Xenoestrogens Affects Breast Cancer Cell Morphology and Motility Relevant for Metastatic Behavior In Vitro.

Breast cancer is highly susceptible to metastasis formation. During the time of disease progression, tumor pathophysiology can be impacted by endogenous factors, like hormonal status, as well as by environmental exposures, such as those related to diet and lifestyle. New lines of evidence point toward a potential role for foodborne endocrine disruptive chemicals in this respect; however, mechanistic understanding remains limited. At the molecular level, crucial steps toward metastasis formation include cell structural changes, alteration of adhesion, and reorganization of cytoskeletal proteins involved in motility. Hence, this study investigates the potential of dietary xenoestrogens to impact selected aspects of breast cancer cell mechanotransduction. Taking the onset of the metastatic cascade as a model, experiments focused on cell-matrix adhesion, single-cell migration, and adaptation of cell morphology. Dietary mycoestrogens alternariol (AOH, 1 μM) and α-zearalenol (α-ZEL, 10 nM), soy isoflavone genistein (GEN, 1 μM), and food packaging plasticizer bisphenol A (BPA, 10 nM) were applied as single compounds or in mixtures. Pursuing the hypothesis that endocrine active molecules could affect cell functions beyond the estrogen receptor-dependent cascade, experiments were performed comparing the MCF-7 cell line to the triple negative breast cancer cells MDA MB-231. Indeed, the four compounds functionally affected the motility and the adhesion of both cell types. These responses were coherent with rearrangements of the actin cytoskeleton and with the modulation of the expression of integrin β1 and cathepsin D. Mechanistically, molecular dynamics simulations confirmed a potential interaction with fragments of the α1 and β1 integrin subunits. In sum, dietary xenoestrogens proved effective in modifying the motility and adhesion of breast cancer cells, as predictive end points for metastatic behavior in vitro. These effects were measurable after short incubation times (1 or 8 h) and contribute to shed novel light on the activity of compounds with hormonal mimicry potential in breast cancer progression.

Integrin profle of circulating tumour cells in breast cancer patients

Background. Integrins, as adhesion molecules, play a key role in the interaction of cells with the basal membrane and intercellular matrix. Numerous studies demonstrate evidence of increased expression of integrins on tumor cells in different types of cancer. Thus, β3 and αV integrins are associated with stem-like features of tumor cells, and β4 integrin as α6β4 heterodimer provides anchorage-independent survival of malignant mammary epithelial cells. However, all the described functions of integrins have been investigated exclusively on primary tumor cells. The functional significance and expression pattern of integrins on circulating tumor cells (CTCs) remains unclear. The aim of the study was to evaluate the β3, β4 and αvβ5 integrin expression on CTCs and its association with molecular subtype, stage and lymph node metastasis in breast cancer patients Material and Methods. The study included 22 patients with T1–4N0–3M0 invasive ductal breast carcinoma. Venous blood was taken from patients without neoadjuvant chemotherapy (group 1) and after neoadjuvant chemotherapy (group 2) in the volume of 12 ml into vacuum tubes with EDTA. The expression of CTC integrins including stemness features CD44/CD24, CD133 and ALDH1, and epithelial-mesenchymal transition (EMT) (N-cadherin) was evaluated by flow cytometry. Results. CTCs with β3+β4-αvβ5- and β3-β4+αvβ5+ phenotypes and stemness properties were associated with larger tumor size (T4) in breast cancer patients. The β3 integrin expression was associated with more aggressive molecular subtypes of breast cancer. Administration of neoadjuvant chemotherapy did not affect the expression pattern of β3, β4 and αvβ5 integrins in CTCs. Conclusion. In breast cancer, most CTCs expressed β3, β4 and αvβ5 integrins despite the lack of attachment to the basal membrane and intercellular matrix. The expression of the above integrins on CTCs was associated with breast cancer molecular subtype, stage and lymph node metastasis, and therefore its evaluation can be considered as one of the objectives of liquid biopsy study.

Abstract 64: Smooth Muscle Cullin-3 Regulates the Renal Baroreceptor Mechanism: a Link Between CUL3 and Integrin β1

Cullin-3 (CUL3) is a critical component of the CUL3-Ring-Ligase (CRL) ubiquitin ligase complex. CUL3 mutants in smooth muscle cells (SMCs) cause vascular dysfunction, arterial stiffness, and severe hypertension (HTN). We hypothesized that conditional deletion of CUL3 in SMCs (S-CUL3KO) led to HTN via 1) a renin-angiotensin system (RAS)-dependent mechanism, and 2) impaired renal baroreceptor mechanism regulating renin expression. CUL3 Flox mice were bred to mice carrying tamoxifen-inducible CRE-recombinase under a smooth muscle promoter (ISM-CRE, control). S-CUL3KO mice exhibited augmented systolic blood pressure (SBP) (ISM-CRE: 124±1 vs. S-CUL3KO: 167±3 mmHg, p&lt;0.05) 3-weeks post tamoxifen. Once HTN was established we used an angiotensin-converting enzyme inhibitor (captopril, 10 mg/kg/day in drinking water) for 1 week to reverse HTN. We observed a greater depressor effect in S-CUL3KO (ΔSBP -49±3 mmHg) compared to ISM-CRE (ΔSBP -33±1 mmHg, p&lt;0.05). A similar effect was identified following angiotensin II type I receptor blocker (ARB, candesartan, 100 mg/kg/day in drinking water) treatment (ΔSBP, ISM-CRE: -27±2 vs. S-CUL3KO: -37±2, p&lt;0.05). We prevented HTN in S-CUL3KO by administering ARB before tamoxifen injection (SBP 120±5 mmHg). Mesenteric arteries exhibited augmented Ang-II induced vasoconstriction, which was prevented with ARB, suggesting that HTN in S-CUL3KO is RAS-dependent and AT 1 R-mediated. Despite severe HTN, S-CUL3KO did not suppress renin mRNA expression in kidneys. We hypothesize that S-CUL3KO exhibits a defective renal baroreceptor mechanism causing impaired suppression of renin expression. We propose that CUL3 participates in the degradation of integrin β1, the key mechanosensory of the renal baroreceptor, via Rab-mediated trafficking and lysosomal fusion. Supporting this, HEK293 cells lacking CUL3 exhibited lower integrin β1 protein expression (0.51±0.08 vs. 0.09±0.01 normalized RFU, p&lt;0.05) concomitant with increased Rab5 and Rab7 expression (3.6±0.3 vs. 1.9±0.1 and 0.95±0.12 vs. 0.37±0.04 normalized RFU). S-CUL3KO demonstrated severely impaired morphological distribution of integrin β1 protein in the juxtaglomerular apparatus. In conclusion, our findings support the hypothesis that 1) CUL3 regulates RAS via the renal baroreceptor, and 2) Rab proteins regulating integrin β1 expression are potential targets for the CRL3 pathway. Understanding CUL3 target proteins might contribute to the development of new strategies to treat HTN.

Iron-laden macrophage-mediated paracrine profibrotic signaling induces lung fibroblast activation.

Idiopathic pulmonary fibrosis (IPF) is a devastating condition characterized by progressive lung scarring and uncontrolled fibroblast proliferation, inevitably leading to organ dysfunction and mortality. Although elevated iron levels have been observed in patients and animal models of lung fibrosis, the mechanisms linking iron dysregulation to lung fibrosis pathogenesis, particularly the role of macrophages in orchestrating this process, remain poorly elucidated. Here we evaluate iron metabolism in macrophages during pulmonary fibrosis using both in vivo and in vitro approaches. In murine bleomycin- and amiodarone-induced pulmonary fibrosis models, we observed significant iron deposition and lipid peroxidation in pulmonary macrophages. Intriguingly, the ferroptosis regulator glutathione peroxidase 4 (GPX4) was upregulated in pulmonary macrophages following bleomycin instillation, a finding corroborated by single-cell RNA sequencing analysis. Moreover, macrophages isolated from fibrotic mouse lungs exhibited increased transforming growth factor (TGF)-β1 expression that correlated with lipid peroxidation. In vitro, iron overload in bone marrow-derived macrophages triggered lipid peroxidation and TGF-β1 upregulation, which was effectively suppressed by ferroptosis inhibitors. When cocultured with iron-overloaded macrophages, lung fibroblasts exhibited heightened activation, evidenced by increased α-smooth muscle actin and fibronectin expression. Importantly, this profibrotic effect was attenuated by treating macrophages with a ferroptosis inhibitor or blocking TGF-β receptor signaling in fibroblasts. Collectively, our study elucidates a novel mechanistic paradigm in which the accumulation of iron within macrophages initiates lipid peroxidation, thereby amplifying TGF-β1 production, subsequently instigating fibroblast activation through paracrine signaling. Thus, inhibiting iron overload and lipid peroxidation warrants further exploration as a strategy to suppress fibrotic stimulation by disease-associated macrophages. NEW & NOTEWORTHY This study investigates the role of iron in pulmonary fibrosis, specifically focusing on macrophage-mediated mechanisms. Iron accumulation in fibrotic lung macrophages triggers lipid peroxidation and an upregulation of transforming growth factor (TGF)-β1 expression. Coculturing iron-laden macrophages activates lung fibroblasts in a TGF-β1-dependent manner, which can be mitigated by ferroptosis inhibitors. These findings underscore the potential of targeting iron overload and lipid peroxidation as a promising strategy to alleviate fibrotic stimulation provoked by disease-associated macrophages.

Effects of Bushen Huoxue on integrin β3 and integrin β5 in the placental tissue of mice with repeated implantation failure caused by phospholipid antibodies.

This study aimed to investigate the effects of the Kidney-Tonifying and Blood-Activating Formula on combating the downregulation of integrin β3 and integrin β5 in mouse placental tissue induced by phospholipid antibodies. This study aimed to investigate the effects of kidney-tonifying and blood-activating formulations on combating the downregulation of integrin β3 and integrin β5 in mouse placental tissue induced by phospholipid antibodies. Mice in the phospholipid antibody group and phospholipid antibody + kidney-tonifying and blood-activating formula group underwent repeated implantation experiments, and the expression of integrin β3 and integrin β5 in placental tissue was observed. The effects of the kidney-tonifying and blood-activating formula on mouse placental tissue were evaluated through biochemical index tests, histopathological observations, and immunohistochemical staining. After intervention with the kidney-tonifying and blood-activating formula, the expression of integrin β3 and integrin β5 in placental tissue was notably upregulated, and placental structure was restored. Kidney-Tonifying and Blood-Activating Formula significantly improved abnormal biochemical indices and exerted a remarkable protective effect on placental tissue pathology. The kidney-tonifying and blood-activating formula effectively counteracted the downregulation of integrin β3 and integrin β5 in mouse placental tissue induced by phospholipid antibodies and alleviated placental tissue pathology through structural improvement and protection. Therefore, kidney-tonifying and blood-activating formulas may serve as potential therapeutic agents for recurrent implantation failure, offering new insights and approaches for clinical treatment.

Abstract We122: Suppression of defender against cell death 1 (Dad1) induces cardiomyocyte death by regulating cell adhesion proteins.

Introduction: Suppression of cardiomyocyte loss is considered a promising strategy for the prevention of the onset of heart failure (HF). Previously, defender against cell death 1 ( Dad1 ) was identified as a cytoprotective gene. Dad1 forms a complex with staurosporine and temperature sensitive 3A (Stt3A), a catalytic subunit of oligosaccharyltransferase (OST) complex which is responsible for N-glycosylation. However, the function of Dad1 in cardiomyocytes remains unknown. Hypothesis: The disruption of cell-extracellular matrix interactions results in cell death, known as anoikis. Because various cell adhesion proteins are N-glycosylated, we hypothesized that Dad1 suppressed anoikis by regulating N-glycosylation of cell adhesion proteins in cardiomyocytes. Methods: Neonatal rat cardiomyocytes were cultured and the expression of Dad1 or Stt3A was knocked down using siRNA. The protein expression was assessed with immunoblot analysis, and cell viability was evaluated using a CellTiter-Blue assay kit. Results: The suppression of Dad1 using siRNAs reduced cardiomyocyte viability ( p &lt;0.01, n=4). Interestingly, suppression of Dad1 impaired the N-glycosylation of integrin β1 and decreased the expression of mature integrin β1 ( p &lt;0.01, n=3). These changes inactivated integrin-mediated signal transduction through dephosphorylation of focal adhesion kinase ( p &lt;0.01, n=5). In addition, enhanced cell-extracellular matrix interactions by using adhesamine rescued the cardiomyocyte death by Dad1 knockdown ( p &lt;0.01, n=4). Next, we pursued the relationship between Dad1 and Stt3A. Dad1 knockdown decreased the expression of Stt3A ( p &lt;0.01, n=6). Suppression of Stt3A induced cardiomyocyte death accompanied by the changes of cell adhesion proteins ( p &lt;0.01, n=4). Finally, double knockdown of Dad1 and Stt3A did not induce a further reduction in cell viability compared with the knockdown of only Dad1 ( p &lt;0.05, n=4), suggesting that cardiomyocyte death by Dad1 knockdown was dependent on Stt3A. Conclusion: Dad1 is required for the stabilization of Stt3A and suppresses cardiomyocyte anoikis by regulating N-glycosylation of cell adhesion proteins in an OST-dependent manner. Dad1-mediated cardiomyocyte survival could be a therapeutic target against HF.

Postoperative radiotherapy with docetaxel versus cisplatin for high-risk oral squamous cell carcinoma: a randomized phase II trial with exploratory analysis of ITGB1 as a potential predictive biomarker

BackgroundOral squamous cell carcinoma (OSCC) causes significant mortality and morbidity worldwide. Surgical resection with adjuvant radiotherapy remains the standard treatment for locally advanced resectable OSCC. Results from landmark trials have established postoperative concurrent cisplatin-radiotherapy (Cis-RT) as the standard treatment for OSCC patients with high-risk pathologic features. However, cisplatin-related toxicity limits usage in clinical practice. Given the need for effective but less toxic alternatives, we previously conducted a single-arm trial showing favorable safety profiles and promising efficacy of concurrent docetaxel-radiotherapy (Doc-RT).MethodsIn this randomized phase 2 trial, we aimed to compare Doc-RT with the standard Cis-RT in postoperative OSCC patients. Eligible patients had AJCC stage III–IV resectable OSCC with high-risk pathologic features. Two hundred twenty-four patients were enrolled and randomly assigned to receive concurrent Doc-RT or Cis-RT. The primary endpoint was 2-year disease-free survival (DFS). Secondary endpoints included overall survival (OS), locoregional-free survival (LRFS), distant metastasis-free survival (DMFS), and adverse events (AEs). Integrin β1 (ITGB1) expression was analyzed as a biomarker for efficacy.ResultsAfter a median 28.8-month follow-up, 2-year DFS rates were 63.7% for Doc-RT arm and 56.1% for Cis-RT arm (p = 0.55). Meanwhile, Doc-RT demonstrated comparable efficacy to Cis-RT in OS, LRFS, and DMFS. Doc-RT resulted in fewer grade 3 or 4 hematological AEs. Low ITGB1 was associated with improved Doc-RT efficacy versus Cis-RT.ConclusionsThis randomized trial directly compared Doc-RT with Cis-RT for high-risk postoperative OSCC patients, with comparable efficacy and less toxicity. ITGB1 merits further validation as a predictive biomarker to identify OSCC patients most likely to benefit from Doc-RT. Findings indicate docetaxel may be considered as a concurrent chemoradiation option in this setting.Trial registrationwww.ClinicalTrials.gov. NCT02923258 (date of registration: October 4, 2016)