tbuckley/cabot-search huggingface dataset:
Poly (I:C)-induced inflammation requires the activation of toll-like receptor 3/Ca<sup>2+</sup>/CaMKII/pannexin 1-dependent signaling
Authors: Magdiel Salgado, Vania Sepúlveda-Arriagada, Macarena Konar-Nié, María A García-Robles, Juan C Sáez
Journal: Theranostics (2025)

Pannexin1 (Panx1) is a glycoprotein, ubiquitously expressed throughout vertebrate tissues. In the cell membrane, it forms non-selective hemichannels (Panx1 HCs) that allow the release of ATP. This extracellular ATP triggers purinergic signaling relevant to the immune responses to pathogens, including viruses. While the activity of Panx1 HCs is known to be elevated by some viruses, the underlying molecular mechanism remains elusive. Methods: In this study, we used Poly(I:C), a double-stranded RNA analog that constitutes a hallmark of viral infections. Peritoneal macrophages were obtained from wild-type and Panx1 knock-out mice. The mRNA levels of proinflammatory cytokines were quantified by RT-qPCR. We also evaluated hemichannel activity through dye uptake assays, whereas Ca2+ signals were studied using Fura-2 and GcamP6. Panx1-P2X7R interaction was studied by proximity ligation assays. Results: Panx1 expression and activity were crucial for the proinflammatory response induced by Poly(I:C) in RAW264.7 cells and peritoneal macrophages. In HeLa cells transfected with mPanx1 (HeLa-mPanx1) and RAW264.7 cells, Poly (I:C) increased Panx1 HC activity in a concentration-dependent manner, which was inhibited by 10Panx1, a peptide that selectively blocks Panx1 HCs. Furthermore, the Poly(I:C)-induced rise in Panx1 HC activity correlated with a rapid increase in intracellular Ca2+ signal, dependent on TLR3 and P2X7R activity. Interestingly, lasting exposure to Poly (I:C) promoted the interaction and internalization of the Panx1-P2X7R complex, which depended on CaMKII, Panx1 HC, and P2X7R activities. The Poly (I:C)-induced increase in Panx1 HC activity was entirely prevented by Ca2+ chelation with BAPTA-AM, CaMKII blockage with KN-62, or PKA activation with db-cAMP. These findings were consistent with data from Panx1 mutants that either avoid or mimic phosphorylation at kinase target sites. Supporting this finding, we demonstrated that CaMKII activity is essential for the inflammatory response triggered by Poly (I:C) in macrophages. Conclusion: A TLR3/Ca2+/CaMKII/Panx1 HC pathway is crucial in orchestrating the cellular response to viral patterns and presents a potential novel target for preventing infections and alleviating the harmful effects associated with RNA-based viral infections.
Autophagy-enhanced nanosonosensitizer mediated sonodynamic therapy for post-myocardial infarction neuromodulation and arrhythmia prevention
Authors: Haoyuan Hu, Songyun Wang, Qian Li, Jiahui Zhao, Yida Pang, Jiale Wang, Huijun Wu, Xinqi Wang, Ye Cheng, Mengran Yu, Xinyue Yin, Yan Zhang, Lilei Yu, Yao Sun, Hong Jiang
Journal: Theranostics (2025)

Rationale: Sympathetic hyperactivation and neuroinflammation are the main triggers of malignant ventricular arrhythmias (VAs) after myocardial infarction (MI). Previous studies proved that photothermal therapy (PTT) and photodynamic therapy (PDT) could reduce MI-induced VAs by inhibiting neuroinflammation. However, the limited penetration depth and potential phototoxicity of phototherapy impose constraints on its further application. As a treatment strategy derived from phototherapy, sonodynamic therapy (SDT) offers exceptional advantages, including excellent penetration capability, temporal-spatial controllability, superior efficacy and minimal side effects. Therefore, it is worthwhile to investigate the effects of sonodynamic modulation on neuroinflammation and arrhythmia prevention. Methods: We designed a long-wavelength emissive sonosensitizer (named BBTD-TPA) based on donor-acceptor-donor scaffold. Subsequently, the compound was encapsulated in DSPE-PEG5000 to form BBTD-TPA nanoparticles (NPs). In vitro experiments were conducted to determine the optimal concentration of BBTD-TPA NPs-mediated SDT and to verify the effects and pathways on autophagy in BV2 cells. The distribution and metabolism of BBTD-TPA NPs in vivo were assessed by NIR-II fluorescence imaging. Finally, in vivo studies were performed to assess the effect of BBTD-TPA NPs-mediated SDT on post-MI sympathetic neuroinflammation and the occurrence of VAs. Results: In vitro studies demonstrated that BBTD-TPA NPs combined with LIFU could promote microglial autophagy via the ROS-AMPK-mTOR pathway. BBTD-TPA NPs were further microinjected into the paraventricular nucleus (PVN), real-time NIR-II fluorescence imaging showed that BBTD-TPA NPs could remain in the PVN for up to 12 h and be metabolized through the liver and kidney. Further in vivo results verified that BBTD-TPA NPs-mediated SDT could inhibit sympathetic nervous activity, and inflammatory responses, thus preventing MI-induced VAs. Conclusion: BBTD-TPA NPs-mediated SDT can promote microglial autophagy and inhibit sympathetic neuroinflammation, thus reducing MI-induced VAs. The current research may inspire a novel strategy for neuromodulation and arrhythmia prevention, providing broader prospects for clinical translation of nanomedical technology.
Ultrasmall radical metal organic cage as cascade antioxidant nanozyme for renal injury
Authors: Cheng Huang, Ziyu Liu, Yucen Deng, Xiaoyan Wang, Qing Miao, Demei Sun, Xinyuan Zhu, Jinghui Yang, Youfu Wang
Journal: Theranostics (2025)

Rationale: As substitutes for natural enzymes, nanozymes offer tunable enzyme-like activities and remarkable structural stability, granting them the potential to treat various diseases, including renal ischemia-reperfusion (I/R) injury. However, the majority of developed nanozymes suffer from unclear structures and limited activity profiles, which hinder the study of their structure-activity relationships, catalytic diversity, mass production, and clinical application. Methods: Herein, we introduce an atomically precise and ultrasmall cascade nanozyme based on a radical-functionalized metal-organic cage (MOC-R). This nanozyme is synthesized through the coordination of radical ligands with copper ions, resulting in a cuboctahedral structure. Results: The MOC-R exhibits cascade antioxidant activities, mimicking the functions of superoxide dismutase (SOD) and catalase (CAT), owing to the synergism between the external radicals and internal copper clusters. The MOC-R nanozyme demonstrates exceptional radical scavenging and anti-inflammatory properties. It mitigates immune cell infiltration, promotes macrophage polarization towards the M2-like phenotype, reduces inflammatory cytokine secretion, and suppresses excessive autophagy and apoptosis. Conclusions: This study not only presents an atomically precise cascade nanozyme but also highlights its promising therapeutic potential for renal I/R injury.
3D bioprintable Mg<sup>2+</sup>-incorporated hydrogels tailored for regeneration of volumetric muscle loss
Authors: Moon Sung Kang, Jeong Min Kim, Hyo Jung Jo, Hye Jin Heo, Yun Hak Kim, Kyung Min Park, Dong-Wook Han
Journal: Theranostics (2025)

Rationale: Current therapeutic approaches for volumetric muscle loss (VML) face challenges owing to limited graft availability and insufficient bioactivity. Three-dimensional (3D) bioprinting has become an alternative technology for fabricating native tissue-mimetic grafts, allowing for tailored structures and complex designs. Methods: We developed an Mg2+-incorporated bioink composed of thiolated gelatin (GtnSH) and maleimide-conjugated gelatin (GtnMI) decorated with magnesium peroxide (MgO2), referred to as a GtnSH/GtnMI/MgO2 bioink. We designed in situ crosslinking between GtnSH and GtnMI to prepare cytocompatible bioink for 3D bioprinting of muscle mimetics. Results: The incorporated MgO2 particles provided oxygen supplementation and myogenic cues. In vitro assays demonstrated that C2C12 myoblasts encapsulated in the GtnSH/GtnMI/MgO2 bioink exhibited high viability, intrinsic proliferation rate, and increased expression of key myogenic markers. In vivo transplantation of the 3D bioprinted GtnSH/GtnMI/MgO2 constructs facilitated muscle mass restoration and M2 macrophage polarization. Additionally, they downregulate the activities of CD4+ and CD8+ lymphocytes, inducing a transition from the initial inflammatory to the restoration phase. Conclusion: The GtnSH/GtnMI/MgO2 bioink is a potential therapeutic strategy for enhancing myogenesis and skeletal muscle tissue regeneration.
Trehalose activates autophagy to alleviate cisplatin-induced chronic kidney injury by targeting the mTOR-dependent TFEB signaling pathway
Authors: Jingchao Yang, Longhui Yuan, Lan Li, Fei Liu, Jingping Liu, Younan Chen, Ping Fu, Yanrong Lu, Yujia Yuan
Journal: Theranostics (2025)

Rationale: Cisplatin is a potent chemotherapeutic agent limited by significant nephrotoxicity. Multiple cycles of cisplatin administration are necessary to confer chronic disease. Autophagy is a lysosomal degradation pathway that enables the clearance and reuse of cytoplasmic components and is essential for maintaining the integrity and normal physiological function of tissues and organs. However, the precise role of autophagy in renal fibrosis has been controversial. Trehalose, a well-known autophagy inducer, plays a cytoprotective role under various stress conditions, such as oxidative damage, dehydration, and temperature changes. In this study, we established a model of cisplatin-induced chronic kidney disease (CKD) and human renal tubular epithelial cells (HK2) injury to investigate the nephroprotective effects of trehalose on cisplatin-induced CKD and the underlying mechanisms involved. Methods: Firstly, we measured the role of autophagy in cisplatin-induced injury models both in vivo and in vitro by western blot and immunofluorescence staining, combined with transcriptomics. Then, biomedical, cellular, and molecular approaches were utilized to evaluate the potential protective effect of trehalose intervention in regulating autophagy. Mechanistically, we performed this study using proximal tubular epithelial cells-specific transcription factor EB (TFEB) knockout mice and TFEB small-interfering RNA technology to determine whether TFEB deficiency affects the pharmacological effected of trehalose in cisplatin-induced injury models. Results: Due to the activation of autophagy, trehalose inhibited mitochondrial dysfunction (mitochondrial fragmentation, depolarization, reactive oxygen species) and cellular senescence induced by cisplatin both in vitro and in vivo. Moreover, renal dysfunction, pathological changes and fibrosis were alleviated in CKD mice after trehalose treatment. Mechanistic investigations revealed that trehalose accumulated in lysosomes and inhibited mTORC1 activity, which triggered TFEB and TFEB-mediated autophagy. In addition, siRNA-mediated knockdown of TFEB in HK2 cells or renal proximal tubular epithelial cells-specific (TECs-specific) TFEB deficiency in mice markedly abolished the beneficial effects of trehalose. Conclusion: Our findings suggested that trehalose induced autophagy to alleviate cisplatin-induced chronic kidney injury by targeting the mTOR-dependent TFEB signaling pathway.
Targeted delivery of Grem1 and IL-10 separately by mesenchymal stem cells effectively mitigates SETD2-deficient inflammatory bowel disease
Authors: Rebiguli Aji, Yue Xu, Ziyi Wang, Wenxin Feng, Liming Gui, Hanyu Rao, Wei Zhang, Ningyuan Liu, Wei-Qiang Gao, Li Li
Journal: Theranostics (2025)

Rationale: Inflammatory bowel disease (IBD) is a relapsing and idiopathic disorder. The low therapeutic efficacy of IBD urgently prompts us to seek new treatment methods. Methods and Results: In this study, we report an adipose-derived mesenchymal stem cell (AT-MSC)-based treatment strategy in which AT-MSCs specifically deliver BMP inhibitor Grem1 and anti-inflammatory factor IL-10 to inflammatory colon tissues in SETD2 deficient dextran sulfate sodium (DSS)-induced colitis mouse models. Targeted delivery of Grem1 reduced colitis by promoting intestinal stem cell regeneration and enhancing mucosal regenerative capacity. Furthermore, targeted delivery of IL-10 reduced colitis by reducing inflammatory cytokines. Conclusion: Our AT-MSCs based therapeutic strategy effectively mitigated IBD. This study has deepened our understanding of IBD therapy and provided a theoretical foundation for its clinical treatment.
Extracellular matrix proteins refine microenvironments for pancreatic organogenesis from induced pluripotent stem cell differentiation
Authors: Ming Hu, Tianzheng Liu, Hui Huang, Derek Ogi, Yinfei Tan, Kaiming Ye, Sha Jin
Journal: Theranostics (2025)

Rationale: The current understanding on manipulating signaling pathways to generate mature human islet organoids with all major hormone-secreting endocrine cell types (i.e., α, β, δ, and γ cells) from induced pluripotent stem cells (iPSCs) is insufficient. However, donor islet shortage necessitates that we produce functional islets in vitro. In this study, we aimed to find decellularized pancreatic extracellular matrix (dpECM) proteins that leverage signaling pathways and promote functional iPSC islet organogenesis. Methods: We performed proteomic analysis to identify key islet promoting factors from porcine and rat dpECM. With this, we identified collagen type II (COL2) as a potential biomaterial cue that endorses islet development from iPSCs. Using global transcriptome profiling, gene set enrichment analysis, immunofluorescence microscopy, flow cytometry, Western blot, and glucose-stimulated hormonal secretion analysis, we examined COL2's role in regulating iPSC pancreatic lineage specification and signaling pathways, critical to islet organogenesis and morphogenesis. Results: We discovered COL2 acts as a functional biomaterial that augments islet development from iPSCs, similar to collagen type V (COL5) as reported in our earlier study. COL2 substantially stimulates the formation of endocrine progenitors and subsequent islet organoids with significantly elevated expressions of pancreatic signature genes and proteins. Furthermore, it enhances islets' glucose sensitivity for hormonal secretion. A cluster of gene expressions associated with various signaling pathways, including but not limited to oxidative phosphorylation, insulin secretion, cell cycle, the canonical WNT, hypoxia, and interferon-γ response, were considerably affected by COL2 and COL5 cues. Conclusion: We demonstrated dpECM's crucial role in refining stem cell differentiation microenvironments for organoid development and maturation. Our findings on biomaterial-stimulated signaling for stem cell specification, organogenesis, and maturation open up a new way to increase the differentiation efficacy of endocrine tissues that can contribute to the production of biologically functional islets.
Nano-fiber/net artificial bionic dura mater promotes neural stem cell differentiating by time sequence external-oral administration to repair spinal cord injury
Authors: Dapeng Zhang, Wenzhao Wang, Shuwei Han, Huiquan Duan, Mengfan Hou, Xiaolong Zhou, Xianzheng Guo, Haosheng Chen, Xiaohong Kong, Xingshuang Zhang, Hengxing Zhou, Shiqing Feng
Journal: Theranostics (2025)

Rationale: The inflammatory microenvironment and resulting neuronal loss following spinal cord injury (SCI) impede the repair process. Endogenous neural stem cells (NSCs), which are constrained by neuroinflammation and slow differentiation rates, are unable to effectively facilitate recovery. Method: In this study, coaxial electrospinning technology was employed to fabricate a nano-fiber/net artificial bionic dura mater (NABDM). This structure featured a core-shell structure, with nafamostat mesylate (NM) encapsulated in the outer layer of poly (lactic-co-glycolic acid) (PLGA) and neurotrophins-3 (NT3) encapsulated in the inner layer of poly (l-lactic acid) (PLLA). Results: The NABDM modifies the polarization direction of microglia in vitro, and promotes the differentiation of NSCs by activating the cGMP-PKG and cAMP signaling pathways. In a mouse SCI model, NABDM effectively reduces local neuroinflammation, accelerates the differentiation of endogenous NSCs, increases the number of mature neurons, and enhances motor, sensory, and autonomic nerve functions in mice. Conclusion: NABDM promotes the differentiation of NSCs and facilitates the repair of SCI in a time sequence external-oral way. This approach represents a rapid and effective new treatment method.
USP14 modulates stem-like properties, tumorigenicity, and radiotherapy resistance in glioblastoma stem cells through stabilization of MST4-phosphorylated ALKBH5
Authors: Xiao Zhou, Qiaoxi Xia, Botao Wang, Junjun Li, Bing Liu, Sisi Wang, Min Huang, Ronghui Zhong, Shi-Yuan Cheng, Xuan Wang, Xiaobing Jiang, Tianzhi Huang
Journal: Theranostics (2025)

Rationale: Glioblastoma (GBM) is the most aggressive type of primary brain cancer and contains self-renewing GBM stem cells (GSCs) that contribute to tumor growth and therapeutic resistance. However, molecular determinants governing therapeutic resistance of GSCs are poorly understood. Methods: We performed genome-wide analysis of deubiquitylating enzymes (DUBs) in patient-derived GSCs and used gene-specific shRNAs to identify an important DUB gene contributing to GSC survival and radioresistance. Subsequently, we employed mass spectrometry and immunoprecipitation to show the interaction between USP14 and ALKBH5, and identified the upstream kinase MST4, which is essential for the deubiquitylation and stabilization of ALKBH5. Additionally, we performed integrated transcriptome and m6A-seq analyses to uncover the key downstream pathways of ALKBH5 that influence GSC radioresistance. Results: Our study demonstrates the essential role of the deubiquitinase USP14 in maintaining the stemness, tumorigenic potential, and radioresistance of GSCs. USP14 stabilizes the m6A demethylase ALKBH5 by preventing its K48-linked ubiquitination and degradation through HECW2. The phosphorylation of ALKBH5 at serine 64 and 69 by MST4 increases its interaction with USP14, promoting ALKBH5 deubiquitylation. Furthermore, ALKBH5 directly interacts with the USP14 transcript in a manner dependent on YTHDF2, establishing a positive feedback loop that sustains the overexpression of both proteins in GSCs. The MST4-USP14-ALKBH5 signaling pathway is crucial for enhancing stem cell-like traits, facilitating homologous recombination repair of DNA double-strand breaks, and promoting radioresistance and tumorigenicity in GSCs. This signaling cascade is further stimulated in GSCs following exposure to ionizing radiation (IR). Inhibiting USP14 with the small molecule IU1 disrupts ALKBH5 deubiquitylation and increases the effectiveness of IR therapy on GSC-derived brain tumor xenografts. Conclusion: Our results identify the MST4-USP14-ALKBH5 signaling pathway as a promising therapeutic target for treating GBM.
The sentinel against brain injury post-subarachnoid hemorrhage: efferocytosis of erythrocytes by leptomeningeal lymphatic endothelial cells
Authors: Hong-Ji Deng, Yun-Huo Xu, Kun Wu, Yun-Cong Li, Yong-Jin Zhang, Han-Fu Yu, Chong Li, Dan Xu, Fei Wang
Journal: Theranostics (2025)

Rationale: The clearance of extravasated erythrocytes represents the most reasonable strategy against brain injury post-subarachnoid hemorrhage (SAH). There is little knowledge about the autologous clearance of extravasated erythrocytes post-SAH. The leptomeningeal lymphatic endothelial cells (LLECs) have been less studied functionally, which were firstly harvested and cultured in vitro by our group previously and are probably related to the clearance of extravasated erythrocytes post-SAH for they closely surround subarachnoid space. Methods: We established a SAH animal model, employed primary LLECs in vitro, mimicked the conditions of the SAH in vitro, performed RNA sequencing, and transfected LLECs with adenovirus and adeno-associated virus both in vivo and in vitro to reveal the molecular mechanisms of efferocytosis of erythrocytes by LLECs and its neuroprotection post-SAH. Results: Firstly, we demonstrated the eryptosis-initiated degradation of extravasated erythrocytes in vitro. Furthermore, we found LLECs preferentially adhered and engulfed apoptotic erythrocytes in vivo and in vitro while sparing from intact erythrocytes, suggesting their novel capacity in the efferocytosis of erythrocytes. Additionally, the efferocytosis of erythrocytes by LLECs plays a role on neuroprotection via improving neurological functions, maintaining neurostructural integrity, and alleviating neuropathological consequences post-SAH. During efferocytosis, phosphatidylserine (PS) and phosphatidylserine receptor (PSR) mediated the recognition of apoptotic erythrocytes by LLECs. We also confirmed that NHL repeat-containing 2 (NHLRC2) positively regulated the efferocytosis of erythrocytes by LLECs to serve as a central regulator in it mediated neuroprotection post-SAH. Conclusions: This study elucidated the efferocytosis of erythrocytes by LLECs and subsequently neuroprotection post-SAH. These findings highlight a prompt, efficient, and regulable pathway for the autologous clearance of extravasated erythrocytes that performs as a sentinel against brain injury post-SAH.
YBX1-driven TUBB6 upregulation facilitates ocular angiogenesis via WNT3A-FZD8 pathway
Authors: Ye-Ran Zhang, Wei-Qi Li, Zhong-Hong Zhang, Ru-Xu Sun, Hong-Jing Zhu, Hui-Ming Qian, Song-Tao Yuan, Yu-Liang Wang
Journal: Theranostics (2025)

Background: Pathological ocular neovascularization, a characteristic feature of proliferative ocular diseases, is a primary contributor to global vision impairment. The dynamics of tubulin are crucial in maintaining ocular homeostasis, closely linked to cellular proliferation and angiogenesis. Elucidating the molecular mechanisms driving this process is vital for formulating effective therapeutic strategies. Methods: Multiple transcriptome analyses revealed upregulation of endothelial tubulin beta-6 chain (Tubb6) in oxygen-induced retinopathy (OIR) and laser-induced choroidal neovascularization (CNV) mice models. Transwell migration assay, wound healing assay, tube formation assay, flow cytometry, and immunofluorescent staining were employed to identify the role of TUBB6 knockout (KO) in vitro. The effects of Tubb6 silencing on retinal angiogenesis and choroidal neovascularization were subsequently evaluated. Results: We identified upregulated Tubb6 expression in retinas from OIR mice through combination analyses of single-cell RNA sequencing (scRNA-Seq) and bulk RNA-Seq. The RNA expression profiles of endothelial cells (ECs) from proliferative diabetic retinopathy (PDR) patients and neovascular age-related macular degeneration (nAMD) patients also exhibited an elevation in TUBB6. Notably, Tubb6 was abundantly expressed in ECs and pericytes, and was predominantly localized to proliferative ECs and vascular tip cells. Functional studies demonstrated that TUBB6 knockdown reduced the expression of proliferative and tip cell markers in vitro. Tubb6 deficiency decreased vascular sprouting and tip cell formation of OIR mice retina and retarded CNV progression in vivo. Mechanistically, YBX1, an RNA-binding protein, was identified as an upstream regulator of TUBB6 via binding to its 3' untranslated region (3'UTR) and maintaining mRNA stability. Transcriptome analysis further linked TUBB6 to the activity of WNT pathway. TUBB6 silencing suppressed the WNT signaling pathway, with WNT3A and FZD8 identified as downstream targets. Conclusions: Collectively, our research shed light on the pivotal function of TUBB6 in maintaining ocular homeostasis and uncovered the YBX1-TUBB6-WNT3A/FZD8 pathway's involvement in sprouting angiogenesis. Targeting TUBB6 and developing its specific inhibitor could pioneer new approaches for treating ocular microvascular diseases.