PDGFRα+ITGA11+ fibroblasts foster early-stage cancer lymphovascular invasion and lymphatic metastasis via ITGA11-SELE interplay.

Cancer-associated fibroblasts (CAFs) exhibit considerable heterogeneity in advanced cancers; however, the functional annotation and mechanism of CAFs in early-stage cancers remain elusive. Utilizing single-cell RNA sequencing and spatial transcriptomic, we identify a previously unknown PDGFRα+ITGA11+ CAF subset in early-stage bladder cancer (BCa). Multicenter clinical analysis of a 910-case cohort confirms that PDGFRα+ITGA11+ CAFs are associated with lymphovascular invasion (LVI) and poor prognosis in early-stage BCa. These CAFs facilitate LVI and lymph node (LN) metastasis in early-stage BCa, as evidenced in a PDGFRα+ITGA11+ CAFs-specific deficient mouse model. Mechanistically, PDGFRα+ITGA11+ CAFs promote lymphangiogenesis via recognizing ITGA11 surface receptor SELE on lymphatic endothelial cells to activate SRC-p-VEGFR3-MAPK pathway. Further, CHI3L1 from PDGFRα+ITGA11+ CAFs aligns the surrounding matrix to assist cancer cell intravasation, fostering early-stage BCa LVI and LN metastasis. Collectively, our study reveals the crucial role of PDGFRα+ITGA11+ CAFs in shaping metastatic landscape, informing the treatment of early-stage BCa LVI.

SUMOylation-Driven mRNA Circularization Enhances Translation and Promotes Lymphatic Metastasis of Bladder Cancer.

Aberrant gene expression is a prominent feature of metastatic cancer. Translational initiation is a vital step in fine-tuning gene expression. Thus, exploring translation initiation regulators may identify therapeutic targets for preventing and treating metastasis. Herein, we identified that DHCR24 was overexpressed in lymph node (LN) metastatic bladder cancer and correlated with poor prognosis of patients. DHCR24 promoted lymphangiogenesis and LN metastasis of bladder cancer in vitro and in vivo. Mechanistically, DHCR24 mediated and recognized the SUMO2 modification at lysine 108 of hnRNPA2B1 to foster TBK1 mRNA circularization and eIF4F initiation complex assembly by enhancing hnRNPA2B1-eIF4G1 interaction. Moreover, DHCR24 directly anchored to TBK1 mRNA 3'-untranslated region to increase its stability, thus forming a feed forward loop to elevate TBK1 expression. TBK1 activated PI3K/Akt signaling to promote VEGFC secretion, resulting in lymphangiogenesis and LN metastasis. DHCR24 silencing significantly impeded bladder cancer lymphangiogenesis and lymphatic metastasis in a patient-derived xenograft model. Collectively, these findings elucidate DHCR24-mediated translation machinery that promotes lymphatic metastasis of bladder cancer and supports the potential application of DHCR24-targeted therapy for LN-metastatic bladder cancer. SIGNIFICANCE: DHCR24 is a SUMOylation regulator that controls translation initiation complex assembly and orchestrates TBK1 mRNA circularization to activate Akt/VEGFC signaling, which stimulates lymphangiogenesis and promotes lymph node metastasis in bladder cancer.

SUMOylation-triggered ALIX activation modulates extracellular vesicles circTLCD4-RWDD3 to promote lymphatic metastasis of non-small cell lung cancer.

Lymph node (LN) metastasis is one of the predominant metastatic routes of non-small cell lung cancer (NSCLC) and is considered as a leading cause for the unsatisfactory prognosis of patients. Although lymphangiogenesis is well-recognized as a crucial process in mediating LN metastasis, the regulatory mechanism involving lymphangiogenesis and LN metastasis in NSCLC remains unclear. In this study, we employed high-throughput sequencing to identify a novel circular RNA (circRNA), circTLCD4-RWDD3, which was significantly upregulated in extracellular vesicles (EVs) from LN metastatic NSCLC and was positively associated with deteriorated OS and DFS of patients with NSCLC from multicenter clinical cohort. Downregulating the expression of EV-packaged circTLCD4-RWDD3 inhibited lymphangiogenesis and LN metastasis of NSCLC both in vitro and in vivo. Mechanically, circTLCD4-RWDD3 physically interacted with hnRNPA2B1 and mediated the SUMO2 modification at K108 residue of hnRNPA2B1 by upregulating UBC9. Subsequently, circTLCD4-RWDD3-induced SUMOylated hnRNPA2B1 was recognized by the SUMO interaction motif (SIM) of ALIX and activated ALIX to recruit ESCRT-III, thereby facilitating the sorting of circTLCD4-RWDD3 into NSCLC cell-derived EVs. Moreover, EV-packaged circTLCD4-RWDD3 was internalized by lymphatic endothelial cells to activate the transcription of PROX1, resulting in the lymphangiogenesis and LN metastasis of NSCLC. Importantly, blocking EV-mediated transmission of circTLCD4-RWDD3 via mutating SIM in ALIX or K108 residue of hnRNPA2B1 inhibited the lymphangiogenesis and LN metastasis of NSCLC in vivo. Our findings reveal a precise mechanism underlying SUMOylated hnRNPA2B1-induced EV packaging of circTLCD4-RWDD3 in facilitating LN metastasis of NSCLC, suggesting that EV-packaged circTLCD4-RWDD3 could be a potential therapeutic target against LN metastatic NSCLC.

ZEB1-mediated biogenesis of circNIPBL sustains the metastasis of bladder cancer via Wnt/ß-catenin pathway.

BACKGROUND: Circular RNAs (circRNAs) circularized by back-splicing of pre-mRNA are widely expressed and affected the proliferation, invasion and metastasis of bladder cancer (BCa). However, the mechanism underlying circRNA biogenesis in mediating the distant metastasis of BCa still unexplored. METHODS: RNA sequencing data between BCa and normal adjacent tissues was applied to identify the differentially expressed circRNAs. The functions of circNIPBL in BCa were investigated via a series of biochemical experiments. The Clinical significance of circNIPBL was examined in a cohort of larger BCa tissues. RESULTS: In the present study, we identified a novel circRNA (hsa_circ_0001472), circNIPBL, which was significantly upregulated and had great influence on the poor prognosis of patients with BCa. Functionally, circNIPBL promotes BCa metastasis in vitro and in vivo. Mechanistically, circNIPBL upregulate the expression of Wnt5a and activated the Wnt/ß-catenin signaling pathway via directly sponged miR-16-2-3p, leading to the upregulation of ZEB1, which triggers the EMT of BCa. Moreover, we revealed that ZEB1 interacted with the flanking introns of exons 2-9 on NIPBL pre-mRNA to trigger circNIPBL biogenesis, thus forming a positive feedback loop. Importantly, circNIPBL overexpression significantly facilitated the distant metastasis of BCa in the orthotopic bladder cancer model, while silencing ZEB1 remarkably blocked the effects of metastasis induced by circNIPBL overexpression. CONCLUSIONS: Our study highlights that circNIPBL-induced Wnt signaling pathway activation triggers ZEB1-mediated circNIPBL biogenesis, which forms a positive feedback loop via the circNIPBL/miR-16-2-3p/Wnt5a/ZEB1 axis, supporting circNIPBL as a novel therapeutic target and potential biomarker for BCa patients.

Biochar performance for preventing cadmium and arsenic accumulation, and the health risks associated with mustard (Brassica juncea) grown in co-contaminated soils.

Cadmium (Cd) and arsenic (As) in co-contaminated soil can enter the human body harming health via the food chain, such as vegetables. Biochar derived from waste has been used to reduce heavy metal uptake by plant, but long-term effects of biochar under Cd and As co-contaminated soil needs to be investigated. A following mustard (Brassica juncea) was grown on co-contaminated soil amended with different raw materials of biochar including biochars pyrolyzed by lignite coal (LCB), rice straw (RSB), silkworm excrement (SEB), and sugar refinery sludge (SSB). The results showed that compared to the control, Cd and As contents of mustard shoot in SSB treatment decreased by 45-49% and 19-37% in two growing seasons, respectively, which was the most effective among 4 biochars. This probably due to SSB owns more abundant Fe-O functional groups. Biochar also altered the microbial community composition, specifically SSB increased proteobacteria abundance by 50% and 80% in the first and second growing seasons, thereby promoted the simultaneous immobilization of Cd and As in soils which may reduce the potential risks to humans. In summary, considering the long-term effects and security of SSB application on mustard, not only is it an effective waste recycle option, but it should also be promoted as a promising approach for safe vegetable production in Cd and As co-contaminated soils.

Effects of foliar applications of Brassinolide and Selenium on the accumulation of Arsenic and Cadmium in rice grains and an assessment of their health risk.

Arsenic and cadmium pose a potential health risk to human beings via rice grain consumption. In the current study, a pot experiment was conducted to evaluate the effect of Br (5 mM and 20 mM) and Se (1 mM) at rice tillering and filling stages on Cd and As accumulation in rice grain and their health risk indices. The results showed that Br or Se applications at different stages of rice improved the photosynthesis, reduce MDA content in flag leaves by 17.41%-38.65%, increased rice biomass and grain yield by 10.50%-29.94% and 10.50%-36.56%, and enhanced grain N and P uptake by 3.25%-34.90%, and 22.98%-72.05%, respectively. Applications of Br and Se effectively decreased Cd and As concentration in rice grain by 31.74%-86.97% and 16.42%-81.13% respectively. Compared to the individual treatment, combined 20 mM Br and 1 mM Se at the filling stage showed the lowest accumulation of As (0.149 mg·kg-1) and Cd (0.105 mg·kg-1) in grain, and its health risk index was below the acceptable limits (HRI < 1). This implies that application of Br and Se at the filling stage is a promising strategy for the safe production of rice in As and Cd co-contaminated regions.

In this study, foliar applications of Br and Se at the grain filling and tillering stage demonstrate their effect on As and Cd accumulation. The findings showed that Br and Se resulted in the Se concentration in grains reaching the Se-enriched level, and the accumulation of As and Cd was the lowest. Furthermore, the application of Br and Se decreased lipid peroxidation, promoted N and P uptake, and increased the rate of photosynthesis in the rice plants, which resulted in increasing rice growth and grain yield. The HRI of heavy metals was below the acceptable limits after application of Br and Se.

Aberrant Nuclear Export of circNCOR1 Underlies SMAD7-Mediated Lymph Node Metastasis of Bladder Cancer.

Circular RNAs (circRNA) containing retained introns are normally sequestered in the nucleus. Dysregulation of cellular homeostasis can drive their nuclear export, which may be involved in cancer metastasis. However, the mechanism underlying circRNA nuclear export and its role in lymph node (LN) metastasis of bladder cancer remain unclear. Here, we identify an intron-retained circRNA, circNCOR1, that is significantly downregulated in LN metastatic bladder cancer and is negatively associated with poor prognosis of patients. Overexpression of circNCOR1 inhibited lymphangiogenesis and LN metastasis of bladder cancer in vitro and in vivo. Nuclear circNCOR1 epigenetically promoted SMAD7 transcription by increasing heterogeneous nuclear ribonucleoprotein L (hnRNPL)-induced H3K9 acetylation in the SMAD7 promoter, leading to inhibition of the TGFß-SMAD signaling pathway. Nuclear retention of circNCOR1 was regulated by small ubiquitin-like modifier (SUMO)ylation of DDX39B, an essential regulatory factor responsible for circRNA nuclear-cytoplasmic transport. Reduced SUMO2 binding to DDX39B markedly increased circNCOR1 retention in the nucleus to suppress bladder cancer LN metastasis. By contrast, SUMOylated DDX39B activated nuclear export of circNCOR1, impairing the suppressive role of circNCOR1 on TGFß-SMAD cascade activation and bladder cancer LN metastasis. In patient-derived xenograft (PDX) models, overexpression of circNCOR1 and inhibition of TGFß signaling significantly repressed tumor growth and LN metastasis. This study highlights SUMOylation-induced nuclear export of circNCOR1 as a key event regulating TGFß-SMAD signaling and bladder cancer lymphangiogenesis, thus supporting circNCOR1 as a novel therapeutic agent for patients with LN metastatic bladder cancer. SIGNIFICANCE: This study identifies the novel intron-retained circNCOR1 and elucidates a SUMOylation-mediated DDX39B-circNCOR1-SMAD7 axis that regulates lymph node metastasis of bladder cancer.

Application of Exogenous Iron Alters the Microbial Community Structure and Reduces the Accumulation of Cadmium and Arsenic in Rice (Oryza sativa L.).

Cadmium (Cd) and arsenic (As) contamination of soil has been a public concern due to their potential accumulation risk through the food chain. This study was conducted to investigate the performance of ferrous sulfate (FeSO4) and ferric oxide (Fe2O3) nanoparticle (Nano-Fe) to stabilize the concentrations of Cd and As in paddy soil. Both Fe treatments led to low extractable Cd and the contents of specifically sorbed As contents, increased (p < 0.05) the Shannon index and decreased (p < 0.05) the Simpson diversity indices compared with the control. Nano-Fe increased the relative abundances of Firmicutes and Proteobacteria and decreased the abundances of Acidobacteria and Chloroflexi. Moreover, the addition of both forms of Fe promoted the formation of Fe plaque and decreased the translocation factor index (TFs) root/soil, TFs shoot/root, and TFs grain/shoot of Cd and As. These results suggest that exogenous Fe may modify the microbial community and decrease the soil available Cd and As contents, inhibit the absorption of Cd and As by the roots and decrease the transport of Cd and As in rice grains and the risk intake in humans. These findings demonstrate that soil amendment with exogenous Fe, particularly Nano-Fe, is a potential approach to simultaneously remediate the accumulation of Cd and As from the soil to rice grain systems.

Cytotoxic effects of polystyrene nanoplastics with different surface functionalization on human HepG2 cells.

Nanoplastics in the environment lead to the human exposure to these particles. However, the consequences of this exposure are not yet fully understood. Here, the cytotoxicity of polystyrene nanoparticles (PS-NPs) with a uniform size (50 nm) but distinct surface functionalization (pristine polystyrene, PS; carboxy and amino functionalized, PS-COOH and PS-NH2, respectively), and at an exposure dosage of 10, 50 and 100 µg/mL, were assessed in the human hepatocellular carcinoma (HepG2) cell line. Although all PS-NPs could be internalized by the HepG2 cells, according to the fluorescent intensities, more of PS-COOH and PS-NH2 than PS, accumulated in the cells. The cell viability was significantly affected in a positively dose-related manner. Functionalized PS-NPs exhibited greater inhibition of cell viability than PS, and the viability inhibition peaked (46%) at 100 µg/mL of PS-NH2 exposure. Superoxide dismutase (SOD) activity was maximum when HepG2 cells were exposed to 10 µg/mL of PS-COOH (1.8 folds higher than that without PS-COOH exposure). The glutathione (GSH) content was maximum when the cells were treated with 50 µg/mL of PS (3.75 fold increase compared to untreated cells). Although the difference in inhibition of cell viability was not significant between PS-NH2 and PS-COOH exposure, 100 µg/mL of PS-NH2 exposure caused the most severe oxidative stress due to dramatically increased accumulation of malondialdehyde (MDA); however, a decrease in the antioxidants levels as the SOD activity and GSH content were also found. The results demonstrated that the cellular oxidative damage occurred and that the antioxidation enzymes may not be able to maintain the balance between the generation of oxidant species and the antioxidant defense. Consequently, 100 µg/mL of PS-NH2 exposure triggered the destruction of antioxidant structures. This study defines the cytotoxic effects of PS-NPs on HepG2 cells and emphasizes the significance of investigating the cytotoxic outcomes of nanoplastics in humans.

Interferon-γ induces the cell surface exposure of phosphatidylserine by activating the protein MLKL in the absence of caspase-8 activity.

Phosphatidylserine (PS), an anionic phospholipid enriched in the inner leaflet of the plasma membrane, is exposed to the outer leaflet during apoptosis. PS exposure was recently shown to be induced during tumor necrosis factor-induced necroptosis. We herein demonstrated that interferon (IFN)-γ induced necroptosis in Caspase-8-knockout mouse-derived embryonic fibroblasts (C8KO MEFs), as well as in WT MEFs co-treated with the pan-caspase inhibitor, z-VAD-fmk. PS exposure and necroptosis were significant after 6- and 24-h treatments with IFN-γ, respectively. To elucidate the molecular mechanisms underlying IFN-γ-induced PS exposure, we generated C8KO MEF-derived cell lines without the expression of RIPK3 (receptor-interacting protein kinase 3), an essential molecule in tumor necrosis factor-induced necroptosis, and IFN-γ-induced PS exposure and necrotic cell death were shown to be specifically inhibited by the loss of RIPK3 expression. Furthermore, the down-regulated expression of MLKL (mixed lineage kinase domain-like protein), a key molecule for inducing membrane rupture downstream of RIPK3 in necroptosis, abolished IFN-γ-induced PS exposure in C8KO MEFs. In human colorectal adenocarcinoma-derived HT29 cells, PS exposure and necroptosis were similarly induced by treatment with IFN-γ in the presence of Smac mimetics and z-VAD-fmk. The removal of IFN-γ from PS-exposing MEFs after a 6-h treatment completely inhibited necroptotic cell death but not the subsequent increase in the number of PS-exposing cells. Therefore, PS exposure mediated by RIPK3-activated MLKL oligomers was induced by a treatment with IFN-γ for a significant interval of time before the induction of necroptosis by membrane rupture.