The integration of precise diagnosis with mechanism-based therapy remains a pivotal challenge in colorectal oncology. Here, building on the triphenylamine (TPA) core, we employed a donor-acceptor (D-A) molecular design strategy by integrating a pyridinium acceptor to engineer TPA-FB, a derivative exhibiting aggregation-induced emission (AIE) characteristic. Both in vitro and in vivo experiments demonstrated that TPA-FB functioned as an effective theranostic agent against colorectal cancer (CRC). Prompted by its molecular structure and notable biological activity, we sought to identify its specific cellular target. We discovered that TPA-FB directly binds to and inhibits the phosphorylation of STAT3, leading to downregulation of SUCLG2. Suppression of SUCLG2 thereby disrupts the tricarboxylic acid (TCA) cycle, reducing succinate and its downstream metabolites, which also disrupts critical protein succinylation modifications. By employing our designed AIEgen to selectively disrupt the STAT3-SUCLG2 axis, we offer a transformative perspective for the development of CRC therapeutics.

Mesenchymal stem cells (MSCs) are a promising cell source for osteoarthritis and intervertebral disc degeneration, but their application is limited by inefficient and unstable chondrogenic differentiation. Here, nine Phorbazole D analogues were synthesized and evaluated for promoting chondrogenesis in human umbilical cord mesenchymal stem cells (hUC-MSCs) under TGF-β/dexamethasone based induction conditions. Among the tested analogues, IO-5a was selected for further evaluation based on its comparatively higher pro-chondrogenic activity and no detectable cytotoxicity. Image-based Alcian blue analysis showed that the positive staining area increased from approximately 17

Tobacco use is the main source of indoor air pollution and contains a variety of toxic components. The smoke from burning cigarettes is a key environmental risk factor that leads to accelerated aging and the occurrence of numerous diseases. Meanwhile, cigarette smoke and aging are both prominent risk factors for age-related macular degeneration (AMD). This study demonstrates that long-term exposure to cigarette smoke can impair retinal function and induce the aging of retinal pigment epithelium (RPE). Meanwhile, the plasma of rats after long-term exposure to cigarette smoke can trigger DNA damage and cellular senescence in vitro. In addition, naphthalene and its metabolites (1,2-dihydroxynaphthalene and 1,2-naphthoquinone) derived from cigarette smoke have been identified as an important factor contributing to RPE damage caused by cigarette exposure. Finally, we found that the aging of RPE induced by smoking can be alleviated through smoking cessation, probably because quitting smoking reduces the accumulation of these toxic chemicals in plasma and within the eyes.

Iron-induced lipid peroxidation of phosphatidylethanolamine (PE) species is a key driver of ferroptosis in retinal pigment epithelial (RPE) cells, a process closely associated with age-related macular degeneration (AMD). The previous studies have demonstrated that induced retinal pigment epithelial (iRPE) cells generated by transcription factor-mediated reprogramming exhibit superior therapeutic efficacy in treating AMD. In this study, it is found that these iRPE cells are resistant to ferroptosis and further identified phosphoethanolamine/phosphocholine phosphatase 1 (PHOSPHO1) as a critical regulator underlying ferroptosis resistance. Mechanistically, PHOSPHO1 inhibits ferroptosis through two distinct mechanisms. First, it reduces PE levels in the endoplasmic reticulum, thereby limiting PE-derived lipid peroxidation. Second, it suppresses autophagy and ferritinophagy, leading to a reduction in intracellular free iron accumulation. Experiments using an in vivo rat model confirm that PHOSPHO1 effectively protects RPE cells from ferroptotic damage. These findings highlight PHOSPHO1 as a potential therapeutic target for AMD, providing insights into novel ferroptosis-based intervention strategies.

The epithelial-mesenchymal transition (EMT) of retinal pigment epithelial (RPE) cells is one of the significant pathogenic mechanisms for the formation of subretinal fibrosis in age-related macular degeneration (AMD). Multiple signaling pathways that promote EMT have been well described, yet the endogenous signaling pathways that inhibit EMT within RPE cells remain largely elusive. In this study, we confirmed the expression of CRX in human RPE cells and human embryonic stem cell-derived RPE (ESC-RPE) cells. By employing sub-culture to disrupt intercellular connections and thereby inhibit the Hippo signaling pathway, combined with TGF-β1 treatment in vitro to mimic the microenvironment for the formation of subretinal fibrosis, it was revealed that Hippo/YAP1 and TGF-β1 synergistically promoted the nuclear translocation of β-catenin, and the latter bound to TCF7 to inhibit the expression of CRX. Overexpression of CRX was capable of suppressing the occurrence of EMT in ESC-RPE cells. CRX exerted its inhibitory effect on EMT partly by upregulating the expression of PPP2R2B. In the laser-induced choroidal neovascularization mouse model, the nuclear translocation of CRX took place in RPE cells, and overexpression of CRX played an inhibitory role in the formation of subretinal fibrosis. This study has identified CRX as an endogenous signaling molecule that inhibits EMT in RPE cells and has provided a new research target and treatment strategy for the treatment of wet AMD and the inhibition of subretinal fibrosis formation.

BackgroundTrihydroxysterols and their analogues accumulate in several pathologies, including neurodegenerative diseases, cancers, and atherosclerosis. Cholestane-3β,5α,6β-triol (CT), recognized as an apoptosis-inducing agent, also exhibits pro-inflammatory effects. Nevertheless, the mechanisms underlying CT-induced cytotoxicity and inflammation remain incompletely characterized.MethodsRNA-sequencing (RNA-seq) analysis indicated CT can stimulate pro-inflammatory cytokine expression. We then employed multiple cell death inhibitors to confirm the predominant form of CT-induced cell death. Using combined chemical inhibition and genetic editing approaches, we established the relationship between caspase 3 activation, CT-mediated gasdermin E (GSDME) cleavage, and subsequent cell death.ResultsCT promotes the expression of multiple pro-inflammatory cytokines. Among inflammatory cell death effector proteins, GSDME was exclusively highly expressed in our cell model. Notably, CT-induced cytotoxicity was abolished by either pharmacological GSDME inhibition or genetic knockdown of GSDME expression. This GSDME-dependent cell death pathway was consistently observed across multiple cell lines. Furthermore, caspase 3 silencing mitigated CT-induced GSDME cleavage, thereby enhancing cell viability.ConclusionCT specifically triggered caspase 3-dependent GSDME cleavage, resulting in pyroptosis as the predominant form of CT-induced cell death. This study establishes a direct mechanistic link between CT and inflammatory cell death execution and provides insight into the contribution of trihydroxysterols to inflammatory pathogenesis.

Cataract, with lens opacity as its feature, often cause vision loss. The main clinical treatment is lens replacement surgery, which usually works well for most of the patients, but not for all. And researching drugs to delay or treat cataracts is also very important socially and scientifically. This study explored the effect of asparagine (Asn) on cataracts. In vivo, a naphthalene-induced cataract model in rats was set up, focusing on lens opacity. In vitro, SRA01/04 cells or cultured lenses were treated with the naphthalene metabolite 1,2-dihydroxynaphthalene (1,2-DHN) to study cellular mechanisms. The results showed that Asn effectively reduced lens opacity in rats with naphthalene-induced cataracts. In vitro experiments revealed that the ATF3/GPX4 signaling pathway is involved in the mechanism by which asparagine inhibits ferroptosis in lens epithelial cells induced by 1,2-DHN, playing a crucial role in this process. When given orally, Asn could cut down the accumulation of ferrous ions caused by naphthalene, stop the production of reactive oxygen species (ROS) and malondialdehyde (MDA), and ease the depletion of glutathione (GSH). In short, our findings suggest that Asn can protect against naphthalene-induced cataracts by reducing ferroptosis. This new discovery surely creates new research directions and strategies for future cataract prevention and treatment.

Supplementary Materials and Methods; Supplementary Figure S1-S11

Localized scleroderma (LoSc) is an autoimmune disease that features extensive fibrosis of the skin. Due to its severity and limited understanding, no effective treatments have been developed to date. Bone marrow mesenchymal stem cells (BMSCs) derived extracellular vesicles (EVs) have been demonstrated promising therapeutic effects on the LoSc mouse model in our previous study. However, identifying the targets and underlying mechanisms of EVs remains a significant challenge for therapeutic applications. miR-143-3p, a critical and abundant factor in BMSC-EVs identified through miRNA sequencing, mediates antifibrotic effects in a LoSc mouse model and is significantly lacking in the dermis of LoSc patients. This microRNA inhibits myofibroblast formation and collagen synthesis, contributing to the therapeutic effects of BMSC-EVs in the LoSc mouse model. Moreover, miR-143-3p-reinforced BMSC-EVs demonstrated enhanced therapeutic efficacy compared to normal BMSC-EVs, reducing dermal thickening, collagen deposition, fibroblast differentiation into myofibroblasts, and promoting skin tissue remodeling. IGF1R, highly expressed in the skin of LoSc, was identified as a potential target of miR-143-3p and was inhibited by miR-143-3p-reinforced EVs, thereby modulating the IGF1/IGF1R-AKT/MAPK pathway. In conclusion, miR-143-3p-enriched EVs could be a more efficient candidate for treating dermal fibrosis in LoSc.

Introduction:Diabetic retinopathy (DR) is one of the most common microvascular complications of diabetes mellitus, and proliferative diabetic retinopathy (PDR) represents its advanced stage. The etiology of PDR is complex. Mitophagy, the selective degradation of dysfunctional mitochondria, is crucial for cellular homeostasis and has been implicated in PDR pathogenesis. However, its specific mechanisms require further investigation. Materials and methods:Gene Expression Omnibu (GEO) datasets (GSE102485, GSE60436) were analyzed in R software to identify differentially expressed mitophagy-related genes (DEMRGs). A PDR diagnostic model was constructed by gene ontology (GO) enrichment analysis, genome enrichment analysis (GSEA), and other relevant methods. Immune infiltration was also performed to analyze the changes in immune cells. Finally, the retinal pigment epithelial cell line (ARPE-19) was incubated with high glucose (HG) to simulate a DR model in vitro, hub-gene expression and mitophagy were assessed by qRT-PCR, Western blotting, and immunofluorescence microscopy (IF). Results:Eight DEMRGs were identified enabling construction of a PDR diagnostic model and prioritization of two hub genes (CASP8 and COL1A1). Finally, qRT-PCR, Western blotting, and IF were performed to provide preliminary validation of the PDR model and HG stimulation increased mitochondria-lysosome colocalization as well as enhanced the expression of mitophagy-related proteins. Conclusion:Integrated bioinformatics and experimental validation suggest that mitophagy contributes to PDR pathogenesis. Five DEMRGs showed up-regulated and immune cell infiltration that may affect the occurrence and PDR development by regulating mitophagy. These findings provide candidate biomarkers and mechanistic insight into PDR.

High hydrogen sulfide (H2S) expression has been shown to play a crucial role in tumor growth and proliferation in colon cancer. So far, most probes used for monitoring H2S offer single-modal imaging, which makes the collection of accurate information for tumor diagnosis difficult. In the present study, a novel dual-modal imaging probe in the second near-infrared (NIR-II, 950-1700) window is proposed and constructed by coating a uniform shell of polydopamine (PDA) on the surface of cuprous oxide (Cu2O)-conjugated downconversion nanoparticles (DCNPs). Of note, DCNP-Cu2O@PDA is in the "on" state, with emission centered at 1550 nm (irradiation using a 980nm laser) in non-neoplastic tissues. Colon tumors with endogenous H2S overexpression can induce the in situ transformation of Cu2O to copper sulfide (Cu2-XS?. Owing to the competitive absorption between Cu2-XS and DCNPs at a wavelength of 980 nm, the generated DCNP-Cu2-XS@PDA exhibits a quenched NIR-II fluorescence (FL) signal. Meanwhile, Cu2-XS exhibits a strong absorbance at 1250 nm, enhancing the NIR-II photoacoustic (PA) signal accompanied by NIR-II FL attenuation. The switching in PA and FL signals at the tumor site offered "double assurances" for the accurate monitoring of H2S. A novel dual-modal imagingprobe in the second near-infrared (NIR-II) window is developed with "double assurances" for the accurate imaging and monitoring of H2S in vivo.image

BACKGROUND:NOP2/Sun domain 2 (NSUN2) is one of the important RNA methyltransferases catalyzing 5-methylcytosine (m5C) formation and participates in many critical bioprocesses. However, the roles and underlying molecular mechanisms of NSUN2-mediated m5C modification in colorectal cancer (CRC) remain unclear.METHODS:To explore the NSUN2 expression in CRC, fresh tissue samples were collected and Nsun2 knockout mouse was constructed. In vitro and in vivo functional assays were conducted to assess the role of NSUN2. RNA array and bisulfite sequencing were used to investigate the potential targets. The mechanisms of NSUN2 function on SKIL were identified by m5C-methylated-RNA immunoprecipitation and RNA stability assays. Additionally, tissue microarray analysis was conducted and patient-derived tumour xenograft mouse (PDX) models were used to define the potential therapeutic targets.RESULTS:NSUN2 was highly expressed in CRC and correlated with poor CRC patient survival. Moreover, silencing NSUN2 suppressed CRC tumourigenesis and progression in Nsun2 knockout mouse models. In vitro and in vivo studies suggested that NSUN2 promoted colorectal cancer cell growth. Mechanistically, SKI-like proto-oncogene (SKIL) is positively regulated by NSUN2, and the NSUN2-SKIL axis is clinically relevant to CRC. NSUN2 induced m5C modification of SKIL and stabilized its mRNA, which was mediated by Y-box binding protein 1 (YBX1). Elevated SKIL levels increased transcriptional coactivator with PDZ-binding motif (TAZ) activation.CONCLUSIONS:Our findings highlight the importance of NSUN2 in the initiation and progression of CRC via m5C-YBX1-dependent stabilization of the SKIL transcript, providing a promising targeted therapeutic strategy for CRC.

Background Colorectal cancer (CRC) is the leading cancer worldwide. Microbial agents have been considered to contribute to the pathogenesis of different disease. But the underlying relevance between CRC and microbiota remain unclear. Methods We dissected the fecal microbiome structure and genomic and transcriptomic profiles of matched tumor and normal mucosa tissues from 41 CRC patients. Of which, the relationship between CRC-associated bacterial taxa and their significantly correlated somatic mutated gene was investigated by exome sequencing technology. Differentially expressed functional genes in CRC were clustered according to their correlation with differentially abundant species, following by annotation with DAVID. The composition of immune and stromal cell types was identified by XCELL. Results We identified a set of 22 microbial gut species associated with CRC and estimate the relative abundance of KEGG ontology categories. Next, the interactions between CRC-related gut microbes and clinical phenotypes were evaluated. 4 significantly mutated gene: TP53, APC, KRAS, SMAD4 were pointed out and the associations with cancer related microbes were identified. Among them, Fusobacterium nucleatum positively corelated with different host metabolic pathways. Finally, we revealed that Fusobacterium nucleatum modified the tumor immune environment by TNFSF9 gene expression. Conclusion Collectively, our multi-omics data could help identify novel biomarkers to inform clinical decision-making in the detection and diagnosis of CRC.

BackgroundGlia maturation factor beta (GMFB) is a growth and differentiation factor that acts as an intracellular regulator of signal transduction pathways. The small ubiquitin-related modifier (SUMO) modification, SUMOylation, is a posttranslational modification (PTM) that plays a key role in protein subcellular localization, stability, transcription, and enzymatic activity. Recent studies have highlighted the importance of SUMOylation in the inflammation and progression of numerous diseases. However, the relationship between GMFB and SUMOylation is unclear.ResultsHere, we report for the first time that GMFB and SUMO1 are markedly increased in retinal pigment epithelial (RPE) cells at the early stage of diabetes mellitus (DM) under hyperglycemia. The GMFΒ protein could be mono-SUMOylated by SUMO1 at the K20, K35, K58 or K97 sites. SUMOylation of GMFB led to its increased protein stability and subcellular translocation. Furthermore, deSUMOylation of GMFΒ downregulates multiple signaling pathways, including the Jak-STAT signaling pathway, p38 pathway and NF-kappa B signaling pathway.ConclusionsThis work provides novel insight into the role of SUMOylated GMFB in RPE cells and provides a novel therapeutic target for diabetic retinopathy (DR).

The permeability of the highly selective blood-brain barrier (BBB) to anticancer drugs and the difficulties in defining deep tumor boundaries often reduce the effectiveness of glioma treatment. Thus, exploring the combination of multiple treatment modalities under the guidance of second-generation near-infrared (NIR-II) window fluorescence (FL) imaging is considered a strategic approach in glioma theranostics. Herein, a hybrid X-ray-activated nanoprodrug was developed to precisely visualize the structural features of glioma microvasculature and delineate the boundary of glioma for synergistic chemo-radiotherapy. The nanoprodrug comprised down-converted nanoparticle (DCNP) coated with X-ray sensitive poly(Se-Se/DOX-co-acrylic acid) and targeted Angiopep-2 peptide (DCNP@P(Se-DOX)@ANG). Because of its ultrasmall size and the presence of DOX, the nanoprodrug could easily cross BBB to precisely monitor and localize glioblastoma via intracranial NIR-II FL imaging and synergistically administer antiglioblastoma chemo-radiotherapy through specific X-ray-induced DOX release and radiosensitization. This study provides a novel and effective strategy for glioblastoma imaging and chemo-radiotherapy.

Real-time monitoring the therapeutic process of sonodynamic therapy (SDT) is essential to optimize the treatment course in time and eventually improve the efficacy. The generation of singlet oxygen (1O2) is a quintessential characteristic of SDT, which permits non-invasive monitoring of SDT by real-time imaging of 1O2 inside the tumor. Nonetheless, the majority of probes are unable to measure 1O2 in real time because of its short half-life and strong oxidative capacity. Here, the study constructs a ratiometric nanoplatform (DTPI) utilizing two fluorescent probes and the sonosensitizer TiO2. The poisonous 1O2 generated by DTPI following ultrasonic (US) radiation efficiently destroys tumor cells. The structural disruption of fluorescent dye IR-1061 by 1O2 leads to a reduction in the DTPI fluorescence signal at 1100 nm, while US radiation has no impact on the fluorescence signal at 1550 nm. Thus, DTPI provides a precise and consistent reflection of the treatment efficacy at the tumor site, leveraging the ratiometric fluorescence signal and variations in oxygen content throughout the treatment process. This ratiometric-fluorescence-based reflection strategy establishes an effective and dependable platform for the real-time monitoring and assessment of the cancer therapeutic effect through ratiometric probes. The study develops a novel second near-infrared (NIR-II) ratiometric fluorescent nanoplatform for real-time monitoring and evaluating cancer sonodynamic therapy efficacy in vivo. image

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Background:Ferroptosis is extremely relevant to the progression of neurodegenerative pathologies such as Alzheimer's disease (AD). Ubiquitin-specific proteases (USP) can affect the NADPH oxidase family. Objective:Our study aimed to elucidate the potential role and molecular basis of a certain USP19 in reducing ferroptosis and mitochondrial injury in AD cells by targeting NOX4 stability. Methods:The deubiquitinase USP family gene USP19, which affects the stability of NOX4 protein, was first screened. The cell model of AD was constructed after interfering with SH-SY5Y cells by Aβ1-40, and then SH-SY5Y cells were infected with lentiviral vectors to knock down USP19 and overexpress NOX4, respectively. Finally, the groups were tested for cell viability, changes in cellular mitochondrial membrane potential, lipid reactive oxygen species, intracellular iron metabolism, and NOX4, Mf1, Mf2, and Drp1 protein expression. Results:5 μmol/L Aβ1-40 intervened in SH-SY5Y cells for 24 h to construct a cell model of AD. Knockdown of USP19 decreased the expression of NOX4 protein, promoted the expression of mitochondrial fusion proteins Mnf1 and Mnf2, and inhibited the expression of the splitting protein Drp1. Furthermore, USP19 knockdown decreased mitochondrial membrane potential, SOD, MDA, intracellular iron content and increased GSH/GSSG ratio in SH-SY5Y cells. Our study revealed that NOX4 protein interacts with USP19 and knockdown of USP19 enhanced ubiquitination to maintain NOX4 protein stability. Conclusions:USP19 attenuates mitochondrial damage in SH-SY5Y cells by targeting NOX4 protein with Aβ1-40.

Cholestane-3β,5α,6β-triol (CT) and its analogues are abundant in natural sources and are reported to demonstrate cytotoxicity toward different kinds of tumor cells without a deep probe into their mechanism of action. CT is also one of the major metabolic oxysterols of cholesterol in mammals and is found to accumulate in various diseases. An extensive exploration of the biological roles of CT over the past few decades has established its identity as an apoptosis inducer. In this study, the effects of CT on A549 cell death were investigated through cell viability assays. RNA-sequencing analysis and western blot of CT-treated A549 cells revealed the role of CT in inducing endoplasmic reticulum (ER) stress response and enhancing autophagy flux, suggesting a putative mechanism of CT-induced cell-death activation involving reactive oxygen species (ROS)-mediated ER stress and autophagy. It is reported for the first time that the upregulation of autophagy induced by CT can serve as a cellular cytotoxicity response in accelerating CT-induced cell death in A549 cells. This research provides evidence for the effect of CT as an oxysterol in cell response to oxidative damage and allows for a deep understanding of cholesterol in its response in an oxidative stress environment that commonly occurs in the progression of various diseases.