Radiation-primed TGF-ß trapping by engineered extracellular vesicles for targeted glioblastoma therapy.

The poor outcome of glioblastoma multiforme (GBM) treated with immunotherapy is attributed to the profound immunosuppressive tumor microenvironment (TME) and the lack of effective delivery across the blood-brain barrier. Radiation therapy (RT) induces an immunogenic antitumor response that is counteracted by evasive mechanisms, among which transforming growth factor-ß (TGF-ß) activation is the most prominent factor. We report an extracellular vesicle (EV)-based nanotherapeutic that traps TGF-ß by expressing the extracellular domain of the TGF-ß type II receptor and targets GBM by decorating the EV surface with RGD peptide. We show that short-burst radiation dramatically enhanced the targeting efficiency of RGD peptide-conjugated EVs to GBM, while the displayed TGF-ß trap reversed radiation-stimulated TGF-ß activation in the TME, offering a synergistic effect in the murine GBM model. The combined therapy significantly increased CD8+ cytotoxic T cells infiltration and M1/M2 macrophage ratio, resulting in the regression of tumor growth and prolongation of overall survival. These results provide an EV-based therapeutic strategy for immune remodeling of the GBM TME and eradication of therapy-resistant tumors, further supporting its clinical translation.

A predictive model for premature atherosclerosis in systemic lupus erythematosus based on clinical characteristics.

OBJECTIVE: Systemic lupus erythematosus (SLE) is associated with a significant risk of atherosclerotic cardiovascular disease, especially in the development of premature atherosclerosis. Specific prediction models for premature atherosclerosis in SLE patients are still limited. The objective of this study was to establish a predictive model for premature atherosclerosis in SLE. METHOD: The study collected clinical and laboratory data from 148 SLE patients under the age of 55, between January 2021 and June 2023. The least absolute shrinkage and selection operator logistic regression model was utilized to identify potentially relevant features. Subsequently, a nomogram was developed using multivariable logistic analysis. The performance of the nomogram was evaluated through a receiver-operating characteristic curve, calibration curve, and decision curve analysis (DCA). RESULTS: A total of 148 SLE patients who fulfilled the inclusion criteria were enrolled in the study, of whom 53 patients (35.81%) met the definition of premature atherosclerosis. Hypertension, antiphospholipid syndrome, azathioprine use, duration of glucocorticoid, and age of patients were included in the multivariable regression. The nomogram, based on the non-overfitting multivariable model, was internally validated and demonstrated sufficient clinical utility for assessing the risk of premature atherosclerosis (area under curve: 0.867). CONCLUSIONS: The comprehensive nomogram constructed in this study serves as a useful and convenient tool for evaluating the risk of premature atherosclerosis in SLE patients. It is helpful for clinicians to early identify SLE patients with premature atherosclerosis and facilitates the implementation of more effective preventive measures. Key Points • SLE patients are at a significantly higher risk of developing premature atherosclerosis compared to the general population, and this risk persists even in cases with low disease activity. Traditional models used to evaluate and predict premature atherosclerosis in SLE patients often underestimate the risk. • This study establishes a comprehensive and visually orientated predictive model of premature atherosclerosis in SLE patients, based on clinical characteristics. • The scoring system allows for convenient and effective prediction of individual incidence of premature atherosclerosis, and could provide valuable information for identification and making further intervention decision.

Optical Coherence Tomography Angiography in Vogt-Koyanagi-Harada Patients: A Systematic Review and Meta-Analysis.

PURPOSE: To describe the choroidal retinal microvascular system in Vogt-Koyanagi-Harada (VKH) subjects and furnish additional proof for the early authentication and treatment of VKH suffers. METHODS: From the beginning to July 2023, a comprehensive search for issued articles on optical coherence tomography angiography (OCTA) among VKH sufferers was implemented in Embase, PubMed, and Web of Science databases. This meta-analysis included 9 eligible studies. Primary endpoints included four kinds of vascular densities, such as superficial capillaris plexus (SCP), deep capillaris plexus (DCP), and choriocapillary (CC). In addition to these, there were foveal avascular zone (FAZ), central retinal thickness (CT), best-corrected distance visual acuity (BCVA log MAR), and subfoveal choroidal thickness (SFCT). RESULTS: SCP and DCP vessel densities in maculas were both smaller in VKH sufferers in the active stage than those normal and remission examinees (SCP vessel density, p < 0.00001, DCP vessel density, p < 0.00001). Compared to remission, CC vascular density was lower during the active phase. (p < 0.00001). SFCT and CT in the active phase exceeded those in normal and remission examinees (all of them p < 0.00001). In terms of the patients with remission, their FAZ was bigger than that in normal subjects. (MD =0.04, p < 0.0001). CONCLUSIONS: Retinal and choroidal microvasculatures are characteristically changed in active VKH patients, which suggests that OCTA can be used as a tool for VKH follow-up.

Morphometric Assessment of Sclera and Ciliary Body in Patients with Noninfectious Anterior Scleritis: A Quantitative Study Using Ultrasound Biomicroscopy.

BACKGROUND: At present, the severity and grade of anterior scleritis are judged mainly based on the area and location of involvement, whether there is necrosis, etc. Quantitative measurement of sclera and surrounding tissues will help to accurately assess the severity of scleritis and provide quantitative indicators for the choice of treatment. METHODS: We retrospectively analyzed the thickness of sclera and ciliary bodies detected by ultrasound biological microscopy (UBM) in noninfectious anterior scleritis patients who subsequently were treated with topical or systemic treatment, and visited our hospital from March 2014 to March 2021. Age- and sex-matched normal individuals were used as controls. RESULTS: A total of 185 patients (50 males and 135 females) with noninfectious anterior scleritis and 84 (31 males and 53 females) controls were included. In patients with noninfectious scleritis, the thickness of sclera and the ciliary body were significantly greater than those in the control group (p < 0.05). Before treatment, the thickness of sclera and the ciliary body in systemic treatment group was significantly higher than that in topical treatment group (p < 0.05). After treatment, both thicknesses of sclera and the ciliary body decreased significantly (p < 0.05). The ratio of ciliary body thickness from the site of inflammation to the normal position was significantly higher in the systemic treatment group than in the topical treatment group. CONCLUSIONS: UBM quantitatively shows a decrease in AST/CBT in patients with anterior scleritis after treatment. The ratio of ciliary body thickness at the site of information to that at the normal position may be a reference for the choice of treatment.

Elevated expression of hsa_circ_0000479 in neutrophils correlates with features of systemic lupus erythematosus.

OBJECTIVE: Accumulating evidence suggests that differentially expressed circular RNAs (circRNAs) play critical roles in immune cells of systemic lupus erythematosus (SLE) patients. Hsa_circ_0000479 has been studied in the field of cancer and infection, whereas seldom studied in autoimmune diseases. The aim of this study was to investigate the role and clinical value of neutrophil hsa_circ_0000479 in SLE. METHODS: The expression levels of hsa_circ_0000479 in both healthy individuals and SLE patients' neutrophils were detected by qPCR and compared with those in peripheral blood mononuclear cells (PBMCs) . In addition, the correlation of hsa_circ_0000479 levels in neutrophils with the clinical and immunological features of SLE patients was also analysed. RESULTS: The expression levels of hsa_circ_0000479 in the patients with SLE were significantly higher in neutrophils than that of PBMCs, and also significantly higher than that in healthy controls (HCs). Moreover, the expression levels of hsa_circ_0000479 in neutrophils were negatively associated with absolute neutrophil count and complement 3 (C3), whereas positively correlated with anti-dsDNA and anti-nucleosome antibodies in SLE. In addition, SLE patients with higher levels of hsa_circ_0000479 demonstrated more several clinical manifestations, including Raynaud's phenomenon, alopecia and leucopenia. CONCLUSIONS: Hsa_circ_0000479 is up-regulated in neutrophils of SLE patients, and is also associated with several important laboratory indicators and clinical manifestations, suggesting that hsa_circ_0000479 in neutrophils was one of probable factors involved in the pathogenesis of SLE with potential clinical value.

Hsa_circ_0000479 was expressed in neutrophils and was considerably higher than that of PBMCs in SLE patients.The neutrophil hsa_circ_0000479 was correlated with laboratory parameters, including NEUT, C3, anti-dsDNA antibodies and AnuA, in addition to being associated with Raynaud's phenomenon, alopecia, and leucopenia in patients with SLE.Hsa_circ_0000479 in neutrophils may play an influential role in SLE patients and will be important to understand the pathogenesis, stratification and treatment in SLE.

Tyro3 receptor tyrosine kinase contributes to pathogenic phenotypes of fibroblast-like synoviocytes in rheumatoid arthritis and disturbs immune cell balance in experimental arthritis.

Rheumatoid arthritis (RA) is a systemic autoimmune disorder characterized by synovitis and joint damage, the underlying causes of which remain unclear. Our prior investigations revealed a notable correlation between the expression of Tyro3 Protein Tyrosine Kinase (Tyro3TK) and the progression of RA. To further elucidate the pathogenic role of Tyro3TK in RA, we analyzed the influence of Tyro3TK on pathogenic phenotypes of RA fibroblast like synoviocyte (FLS) in vitro and compared disease severity, joint damages and immunological parameters of K/BxN serum transfer arthritis (STA) in Tyro3TK-/- deficient mice and wild type controls. Our findings underscored the remarkable effectiveness of Tyro3TK blockade, as evidenced by diminished secretion of inflammatory cytokines and matrix metalloproteinases (MMPs), curtailed migration and invasiveness of RAFLS, and attenuated differentiation of pathogenic helper T cell subsets mediated by RAFLS. Correspondingly, our in vivo investigations illuminated the more favorable outcomes in Tyro3TK-deficient mice, characterized by reduced joint pathology, tempered synovial inflammation, and restored immune cell equilibrium. These data suggested that Tyro3TK might contribute to aggravated autoimmune arthritis and immunological pathology and act as a potential therapeutic target for RA.

OPTICAL COHERENCE TOMOGRAPHY CHARACTERISTICS OVER TIME IN BEHÇET'S UVEITIS.

PURPOSE: To analyze the characteristics of macular optical coherence tomography in patients with Behçet's uveitis (BU). METHODS: Retrospectively analyzing optical coherence tomography images and clinical data of patients with Behçet's uveitis who visited our hospital from January 2010 to July 2022. RESULTS: One hundred and one patients (174 eyes) were included. We analyzed the development of optical coherence tomography changes in these patients and its relationship with acuity vision and found that cystic macular edema, hyperreflexive retinal spots, inner nuclear layer edema, and outer nuclear layer edema appeared at any time during the disease course. Epiretinal membranes appeared beginning 1 to 2 weeks after onset and worsened over time, and foveal atrophy occurred beginning at 2 to 4 weeks. Foveal atrophy, foveal layers disappear, ellipsoid zone (EZ) disruption, retinal pigment epithelium (RPE) disruption, RPE hyperreflection, and choroidal hyperreflection were correlated with visual acuity. Kaplan‒Meier survival analysis showed at 60 months of follow-up, and almost all patients with foveal atrophy, EZ disruption, RPE disruption, RPE hyperreflection, and choroidal hyperreflection had visual acuity less than logMAR 1.0. EZ disruption and RPE disruption were reversible with aggressive treatment from 4 to 12 months after onset, but was less recoverable after 2 years. In advanced stages, optical coherence tomography manifested as structural disturbances and atrophy of the macular area, highly reflective material deposition in the RPE layer, and a thick macular epimembrane. CONCLUSION: Optical coherence tomography showed that severe lesions occurred in macula of patients with Behçet's uveitis in early stage. Aggressive treatment can partially reverse it.

Immune Checkpoint Inhibition in GBM Primed with Radiation by Engineered Extracellular Vesicles.

The lack of safe and effective delivery across the blood-brain barrier and the profound immune suppressive microenvironment are two main hurdles to glioblastoma (GBM) therapies. Extracellular vesicles (EVs) have been used as therapeutic delivery vehicles to GBM but with limited efficacy. We hypothesized that EV delivery to GBM can be enhanced by (i) modifying the EV surface with a brain-tumor-targeting cyclic RGDyK peptide (RGD-EV) and (ii) using bursts of radiation for enhanced accumulation. In addition, EVs were loaded with small interfering RNA (siRNA) against programmed cell death ligand-1 (PD-L1) for immune checkpoint blockade. We show that this EV-based strategy dramatically enhanced the targeting efficiency of RGD-EV to murine GBM, while the loaded siRNA reversed radiation-stimulated PD-L1 expression on tumor cells and recruited tumor-associated myeloid cells, offering a synergistic effect. The combined therapy significantly increased CD8+ cytotoxic T cells activity, halting tumor growth and prolonging animal survival. The selected cell source for EVs isolation and the presented functionalization strategy are suitable for large-scale production. These results provide an EV-based therapeutic strategy for GBM immune checkpoint therapy which can be translated to clinical applications.

Targeted delivery of neural progenitor cell-derived extracellular vesicles for anti-inflammation after cerebral ischemia.

Ischemic stroke remains a major cause of death, and anti-inflammatory strategies hold great promise for preventing major brain injury during reperfusion. In the past decade, stem cell-derived extracellular vesicles (EVs) have emerged as novel therapeutic effectors in immune modulation. However, the intravenous delivery of EVs into the ischemic brain remains a challenge due to poor targeting of unmodified EVs, and the costs of large-scale production of stem cell-derived EVs hinder their clinical application. Methods: EVs were isolated from a human neural progenitor cell line, and their anti-inflammatory effects were verified in vitro. To attach targeting ligands onto EVs, we generated a recombinant fusion protein containing the arginine-glycine-aspartic acid (RGD)-4C peptide (ACDCRGDCFC) fused to the phosphatidylserine (PS)-binding domains of lactadherin (C1C2), which readily self-associates onto the EV membrane. Subsequently, in a middle cerebral artery occlusion (MCAO) mouse model, the RGD-C1C2-bound EVs (RGD-EV) were intravenously injected through the tail vein, followed by fluorescence imaging and assessment of proinflammatory cytokines expression and microglia activation. Results: The neural progenitor cell-derived EVs showed intrinsic anti-inflammatory activity. The RGD-EV targeted the lesion region of the ischemic brain after intravenous administration, and resulted in a strong suppression of the inflammatory response. Furthermore, RNA sequencing revealed a set of 7 miRNAs packaged in the EVs inhibited MAPK, an inflammation related pathway. Conclusion: These results point to a rapid and easy strategy to produce targeting EVs and suggest a potential therapeutic agent for ischemic stroke.

Fabrication of alignment polycaprolactone scaffolds by combining use of electrospinning and micromolding for regulating Schwann cells behavior.

In the present study, a new approach for fabricating micropatterned polycaprolactone (PCL) scaffolds with ridge/groove structure on the surface was developed by combining use of electrospinning and micromolding method. A series of physicochemical properties, including morphology, wettability, component, crystal pattern and mechanical properties, of prepared PCL scaffolds were characterization, respectively. Stability of the micropatterned PCL scaffolds was measured using phosphate buffer solution immersion for a certain period. Then, the regulating effects of the micropatterned PCL scaffolds on attachment, orientation and normal biological function of Schwann cells were evaluated. And the protein adsorption behavior in various PCL scaffolds was also detected. The results showed that the micropatterned PCL scaffolds demonstrated a porous micro/nano complex structure with enhanced hydrophobicity and mechanical properties as a function of electrospun flow-rate of PCL solution. The micropatterned PCL scaffolds possessed good stability and could effectively regulate the attachment and orientation of Schwann cells at the early stage after cell culture. Importantly, the electrospun flow-rate of PCL solution was found to play an important role in scaffold properties, cell behavior and protein adsorption. The micropatterned scaffolds with a flow-rate of PCL solution at 0.12 mL h-1 demonstrated the better regulation on Schwann cells attachment and alignment without negatively affect the normal biological function of the cells. To the best of our knowledge, this is the first report of combining use of electrospinning and micromolding method for preparing artificial nerve implants. The study is anticipated to have potential application in peripheral nerve and other tissue engineering. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 3123-3134, 2018.