The “double‐edged sword” effect of macrophages under the influence of different microenvironments determines the outcome and prognosis of tissue injury. Accurate and stable reprogramming macrophages (Mφ) are the key to rapid wound healing. In this study, an immunized microsphere‐engineered GelMA hydrogel membrane is constructed for oral mucosa treatment. The nanoporous poly(lactide‐co‐glycolide) (PLGA) microsphere drug delivery system combined with the photo‐cross‐linkable hydrogel is used to release the soybean lecithin (SL)and IL‐4 complexes (SL/IL‐4) sustainedly. In this way, it is realized effective wound fit, improvement of drug encapsulation, and stable triphasic release of interleukin‐4 (IL‐4). In both in vivo and in vitro experiments, it is demonstrated that the hydrogel membrane can reprogram macrophages in the microenvironment into M2Mφ anti‐inflammatory types, thereby inhibiting the local excessive inflammatory response. Meanwhile, high levels of platelet‐derived growth factor (PDGF) secreted by M2Mφ macrophages enhanced neovascular maturation by 5.7‐fold, which assisted in achieving rapid healing of oral mucosa. These findings suggest that the immuno‐engineered hydrogel membrane system can re‐modulating the biological effects of Mφ, and potentiating the maturation of neovascularization, ultimately achieving the rapid repair of mucosal tissue. This new strategy is expected to be a safe and promising immunomodulatory biomimetic material for clinical translation.
Background Glioblastomas (GBMs) are grade IV central nervous system tumors characterized by a poor prognosis and a short median overall survival. Effective induction of GBM cell death is difficult because the GBM cell population is genetically unstable, resistant to chemotherapy and highly angiogenic. In recent studies, ubiquitin-specific protease 7 (USP7) is shown to scavenge ubiquitin from oncogenic protein substrates, so effective inhibition of USP7 may be a potential key treatment for GBM. Methods Immunohistochemistry and western blotting were used to detect the expression of USP7 in GBM tissues. In vitro apoptosis assay of USP7 inhibition was performed by western blotting, immunofluorescence, and flow cytometry. Anti-apoptotic substrates of USP7 were defined by Co-IP and TMT proteomics. Western blotting and IP were used to verify the relationship between USP7 and its substrate. In an in vivo experiment using an intracranial xenograft model in nude mice was constructed to assess the therapeutic effect of target USP7. Results Immunohistochemistry and western blotting confirmed that USP7 was significantly upregulated in glioblastoma samples. In in vitro experiments, inhibition of USP7 in GBM induced significant apoptosis. Co-IP and TMT proteomics identified a key anti-apoptotic substrate of USP7, ADP-ribosylation factor 4 (ARF4). Western blotting and IP confirmed that USP7 interacted directly with ARF4 and catalyzed the removal of the K48-linked polyubiquitinated chain that binded to ARF4. In addition, in vivo experiments revealed that USP7 inhibition significantly suppressed tumor growth and promoted the expression of apoptotic genes. Conclusions Targeted inhibition of USP7 enhances the ubiquitination of ARF4 and ultimately mediates the apoptosis of GBM cells. In a clinical sense, P5091 as a novel specific inhibitor of USP7 may be an effective approach for the treatment of GBM.
Background: Glioblastomas (GBMs) are grade IV central nervous system tumors characterized by a poor prognosis and a median overall survival of 15 months. The glioblastoma cell population is more genetically unstable, resistant to chemotherapy, more angiogenic, and more malignant. Because ubiquitin-specific proteases (USPs) remove ubiquitin from oncogenic protein substrates, effective inhibition of ubiquitin-specific protease 7 (USP7), which is highly expressed in GBMs, is a potentially critical therapeutic approach. Methods: Immunohistochemistry and Western blotting were used to detect high expression of USP7 in GBM. in vitro studies were performed by Western blotting, immunofluorescence, and flow cytometry to detect apoptosis following inhibition of USP7. Anti-apoptotic substrates of USP7 were defined by Co-IP and TMT proteomics. Nude mouse intracranial xenograft models were constructed to verify whether inhibition of USP7 inhibited the proliferation rate of tumours.Results: USP7 is significantly upregulated in glioblastoma samples. Interfering with USP7 in GBMs induced significant apoptosis, which also occurred after treatment with P5091, a novel small molecule inhibitor of USP7. Mechanistically, apoptosis of GBMs after interference with USP7 function is achieved by stabilizing a key anti-apoptotic protein, ADP-ribosylation factor 4 (ARF4).Furthermore, USP7 interacts directly with ARF4 and catalyzes the removal of the K48-linked polyubiquitinated chain that binds to ARF4, thereby stabilizing the protein. In in vivo experiments, P5091 significantly inhibited tumor growth and promoted the expression of apoptotic genes. Conclusion: Targeted inhibition of USP7 enhances the ubiquitination of ARF4 and ultimately mediates the apoptosis of GBM cells. In a clinical sense, P5091 as a specific inhibitor of USP7 may be an effective approach for the treatment of GBM.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.