High-throughput screening identifies ibuprofen as an sEV PD-L1 inhibitor for synergistic cancer immunotherapy.

Programmed death-ligand 1 (PD-L1) on tumor-derived small extracellular vesicles (sEVs) limits therapeutic effectiveness by interacting with the PD-1 receptor on host immune cells. Targeting the secretion of sEV PD-L1 has emerged as a promising strategy to enhance immunotherapy. However, the lack of small-molecule inhibitors poses a challenge for clinical translation. In this study, we developed a target and phenotype dual-driven high-throughput screening strategy that combined virtual screening with nanoflow-based experimental verification. We identified ibuprofen (IBP) as a novel inhibitor that effectively targeted sEV PD-L1 secretion. IBP disrupted the biogenesis and secretion of PD-L1+ sEVs in tumor cells by physically interacting with a critical regulator of sEV biogenesis, hepatocyte growth factor-regulated tyrosine kinase substrate. Notably, the mechanism of action of IBP is distinct from its commonly known targets, cyclooxygenases. Administration of IBP stimulated antitumor immunity and enhanced the efficacy of anti-PD-1 therapy in melanoma and oral squamous cell carcinoma mouse models. To address potential adverse effects, we further developed an IBP gel for topical application, which demonstrated remarkable therapeutic efficacy when combined with anti-PD-1 treatment. The discovery of this specific small inhibitor provides a promising avenue for establishing durable, systemic antitumor immunity.

Overexpression of RAB27A in Oral Squamous Cell Carcinoma Promotes Tumor Migration and Invasion via Modulation of EGFR Membrane Stability.

Oral squamous cell carcinoma (OSCC) is the most prevalent subtype of head and neck tumors, highly prone to lymph node metastasis. This study aims to examine the expression pattern of Ras-related protein Rab-27A (RAB27A) and explore its potential implications in OSCC. The expression of RAB27A was assessed through immunohistochemical analysis utilizing tissue microarrays. In vitro experiments were conducted using RAB27A-knockdown cells to investigate its impact on OSCC tumor cells. Additionally, transcriptome sequencing was performed to elucidate potential underlying mechanisms. RAB27A was significantly overexpressed in OSCC, and particularly in metastatic lymph nodes. It was positively correlated with the clinical progression and poor survival prognosis. Silencing RAB27A notably decreased the proliferation, migration, and invasion abilities of OSCC cells in vitro. A Gene Ontology (GO) enrichment analysis indicated a strong association between RAB27A and the epidermal growth factor receptor (EGFR) signaling pathway. Further investigations revealed that RAB27A regulated the palmitoylation of EGFR via zinc finger DHHC-type containing 13 (ZDHHC13). These findings provide insights into OSCC progression and highlight RAB27A as a potential therapeutic target for combating this aggressive cancer.

Endothelial cells induce degradation of ECM through enhanced secretion of MMP14 carried on extracellular vesicles in venous malformation.

Venous malformations (VMs), featuring localized dilated veins, are the most common developmental vascular anomalies. Aberrantly organized perivascular extracellular matrix (ECM) is one of the prominent pathological hallmarks of VMs, accounting for vascular dysfunction. Although previous studies have revealed various proteins involved in ECM remodeling, the detailed pattern and molecular mechanisms underlying the endothelium-ECM interplay have not been fully elucidated. Our previous studies revealed drastically elevated extracellular vesicle (EV) secretion in VM lesions. Here, we identified increased EV-carried MMP14 in lesion fluids of VMs and culture medium of TIE2-L914F mutant endothelial cells (ECs), along with stronger ECM degradation. Knockdown of RAB27A, a required regulator for vesicle docking and fusion, led to decreased secretion of EV-carried MMP14 in vitro. Histochemical analysis further demonstrated a highly positive correlation between RAB27A in the endothelium and MMP14 in the perivascular environment. Therefore, our results proved that RAB27A-regulated secretion of EV-MMP14, as a new pattern of endothelium-ECM interplay, contributed to the development of VMs by promoting ECM degradation.

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Effective microorganisms (EM) might alleviate deterioration of soil environmental quality and yield decline of pepper (Capsicum annuum) caused by continuous replanting and imbalanced fertilizer application in Xinjiang. We investigated the effects of applying EM microbial agent on the growth of pepper plants, yield, soil nutrient content, soil enzyme activity, and rhizosphere eukaryotic community. The results showed that the application of EM microbial agent increased plant height, stem diameter, leaf length, leaf width and root length by 22.6%, 35.3%, 33.3%, 29.7% and 15.1%, respectively. It also increased fruit width, individual fruit weight, and yield by 5.3%, 42.9%, and 74.7%, respectively. After the application of EM microbial agent, the levels of soil available nitrogen increased by 10.2% and 5.8% during the flowering and maturity stages, respectively. Similarly, available phosphorus increased by 10.4% and 13.4%, respectively. The soil sucrase activity was increased by 40.7%, 14.6%, and 9.3% during the seedling, flowering, and maturity stages, respectively. Urease activity was also increased by 7.9%, 10.2%, and 11.5%, respectively. Furthermore, the application of EM microbial agent increased soil peroxidase activity by 16.8% and 44.6% at flowering and maturity stages, respectively. The application of microbial agent significantly altered the ß-diversity of the rhizosphere eukaryotic community in pepper plants. Specifically, microbial agent increased the relative abundances of populations belonging to Enchytraeus and Sminthurides genera, which could contribute to soil improvement and nutrient cycling. Compared to the CK, the relative abundance of pathogenic microorganisms including Olpidium and Aplanochytrium genera decreased by 98.0% and 89.3%, and the relative abundance of the Verticillium decreased to 0. These results demonstrated that EM microbial agent could increase soil nutrient content, enhance soil enzyme activity, and reduce soil pathogenic fungi in the pepper cultivation areas of Xinjiang, thus achieving beneficial effects on pepper growth and fruit yield.

HRS mediates tumor immune evasion by regulating proteostasis-associated interferon pathway activation.

By sorting receptor tyrosine kinases into endolysosomes, the endosomal sorting complexes required for transport (ESCRTs) are thought to attenuate oncogenic signaling in tumor cells. Paradoxically, ESCRT members are upregulated in tumors. Here, we show that disruption of hepatocyte growth factor-regulated tyrosine kinase substrate (HRS), a pivotal ESCRT component, inhibited tumor growth by promoting CD8+ T cell infiltration in melanoma and colon cancer mouse models. HRS ablation led to misfolded protein accumulation and triggered endoplasmic reticulum (ER) stress, resulting in the activation of the type I interferon pathway in an inositol-requiring enzyme-1α (IRE1α)/X-box binding protein 1 (XBP1)-dependent manner. HRS was upregulated in tumor cells with high tumor mutational burden (TMB). HRS expression associates with the response to PD-L1/PD-1 blockade therapy in melanoma patients with high TMB tumors. HRS ablation sensitized anti-PD-1 treatment in mouse melanoma models. Our study shows a mechanism by which tumor cells with high TMB evade immune surveillance and suggests HRS as a promising target to improve immunotherapy.

HRS Regulates Small Extracellular Vesicle PD-L1 Secretion and Is Associated with Anti-PD-1 Treatment Efficacy.

PD-L1 localized to immunosuppressive small extracellular vesicles (sEV PD-L1) contributes to tumor progression and is associated with resistance to immune-checkpoint blockade (ICB) therapy. Here, by establishing a screening strategy with a combination of tissue microarray (TMA), IHC staining, and measurement of circulating sEV PD-L1, we found that the endosomal sorting complex required for transport (ESCRT) member protein hepatocyte growth factor-regulated tyrosine kinase substrate (HRS) was the key regulator of circulating sEV PD-L1 in head and neck squamous cell carcinoma (HNSCC) patients. Increased HRS expression was found in tumor tissues and positively correlated with elevated circulating sEV PD-L1 in patients with HNSCC. The expression of HRS was also negatively correlated to the infiltration of CD8+ T cells. Knockdown of HRS markedly reduced PD-L1 expression in HNSCC cell-derived sEVs, and these sEVs from HRS knockdown cells showed decreased immunosuppressive effects on CD8+ T cells. Knockout of HRS inhibited tumor growth in immunocompetent mice together with PD-1 blockade. Moreover, a higher HRS expression was associated with a lower response rate to anti-PD-1 therapy in patients with HNSCC. In summary, our study reveals HRS, the core component of ESCRT-0, regulates sEV PD-L1 secretion, and is associated with the response to ICB therapy in patients with HNSCC, suggesting HRS is a promising target to improve cancer immunotherapy.

Targeted inhibition of tumor-derived exosomes as a novel therapeutic option for cancer.

Mounting evidence indicates that tumor-derived exosomes (TDEs) play critical roles in tumor development and progression by regulating components in the tumor microenvironment (TME) in an autocrine or paracrine manner. Moreover, due to their delivery of critical molecules that react to chemotherapy and immunotherapy, TDEs also contribute to tumor drug resistance and impede the effective response of antitumor immunotherapy, thereby leading to poor clinical outcomes. There is a pressing need for the inhibition or removal of TDEs to facilitate the treatment and prognosis of cancer patients. Here, in the present review, we systematically overviewed the current strategies for TDE inhibition and clearance, providing novel insights for future tumor interventions in translational medicine. Moreover, existing challenges and potential prospects for TDE-targeted cancer therapy are also discussed to bridge the gaps between progress and promising applications.

How Do Environmental Concerns and Governance Performance Affect Public Environmental Participation: A Case Study of Waste Sorting in Urban China.

Environmental pollution threatens public health and has become a social concern in recent years. Despite the conditions for public participation in environmental governance have improved considerably, the level of public engagement in government projects still falls short of expectations. Therefore, this article introduced two key variables, hoping to answer the following research question that how environmental concerns and governance performance affect public environmental participation. Through principal component analysis of the data from the "Survey of Chinese Urban Residents' Attitudes toward Environmental Protection", the findings of this article are as follows: First, public environmental concerns have no significant impact on their environmental engagement; second, the improvement of residents' confidence in the government performance of environmental management reduces their willingness to participate in official projects. The higher the confidence in the government's performance, the lower the level of public engagement is. Moreover, due to the consideration of self-interest or lack of environmental awareness, those who oppose waste incineration in waste terminal disposal tend to take a non-participatory role in waste sorting programs. Therefore, we suggest that the government have more diverse shareholders in environmental protection, so it should expand public participation through education, publicity, mobilization, and incentives.

Coexpression of VHb and MceG genes in Mycobacterium sp. Strain LZ2 enhances androstenone production via immobilized repeated batch fermentation.

Androstenone production is limited by low-efficiency substrate transport and dissolved oxygen levels during fermentation. In this study, the coexpression of the optimized Vitreoscilla hemoglobin (VHb) and sterol transporter ATPase (MceG) genes in Mycobacterium sp. LZ2 (Msp) was investigated to alleviate dissolved oxygen and mass transfer limitations. Results revealed that Msp-vgb/mceG effectively improved the growth, production, and adaptation to dissolved oxygen compared with those of Msp. The increased catalase activity and reduced intracellular ROS levels enhanced cell viability and promoted transcription of genes critical for phytosterol metabolism. Bagasse as an immobilization carrier increased the productivity of Msp-vgb/mceG by 56%. Immobilized repeat batch fermentation reduced the biotransformation period from 60 days to 37 days and improved the productivity from 0.039 g/L/h to 0.069 g/L/h. To the best of our knowledge, this work is the first study on the immobilization of recombinant mycobacteria on bagasse for androstenone production.

Cloning, expression and characterization of metalloproteinase HypZn from Aspergillus niger.

A predicted metalloproteinase gene, HypZn, was cloned from Aspergillus niger CGMCC 3.7193 and expressed in Pichia pastoris GS115, and the physicochemical characteristics of recombinant HypZn were investigated after separation and purification. The results showed that the specific activity of the purified HypZn reached 1859.2 U/mg, and the optimum temperature and pH value of HypZn were 35°C and 7.0, respectively. HypZn remained stable both at 40°C and at pH values between 5.0 and 8.0. The preferred substrate of HypZn was soybean protein isolates, and the Km and Vmax values were 21.5 µmol/mL and 4926.6 µmol/(mL∙min), respectively. HypZn was activated by Co2+ and Zn2+ and inhibited by Cu2+ and Fe2+. The degree of soybean protein isolate hydrolysis reached 14.7%, and the hydrolysates were of uniform molecular weight. HypZn could tolerate 5000 mM NaCl and completely lost its activity after 30 min at 50°C. The enzymological characterizations indicated that HypZn has great application potential in the food industry, especially in fermented food processing.

Evaluation of the Low Inclusion of Full-Fatted Hermetia illucens Larvae Meal for Layer Chickens: Growth Performance, Nutrient Digestibility, and Gut Health.

Substitution of feed protein source with defatted black soldier fly larvae meal (BSFM) has been evaluated intensively in poultry, but information about full-fatted BSFM is still very limited. The aim of the present study was to investigate the effect of dietary low inclusion of full-fatted BSFM on the growth performance, plasma antioxidant ability, nutrient digestibility, and gut health of layer chickens during 1-42 days of age. A total of 480 female 1-day-old Hy-Line Brown chickens were divided into four dietary treatments, with the inclusion of 0, 3, 6, and 9% of full-fatted BSFM. Each treatment included six replicates and 20 birds per replicate. As dietary full-fatted BSFM inclusion levels increased, there was a quadratic increase in final weight and average daily gain and a quadratic decrease in feed/gain ratio. Dietary full-fatted BSFM inclusion levels increased the digestibility of crude protein and ether extract quadratically as well as ileum mucosal sIgA concentration linearly, but these had no effect on intestinal morphology. Additionally, an increase in dietary full-fatted BSFM inclusion levels resulted in a linear increase in glutathione peroxidase and total superoxide dismutase activities and a linear decrease in malondialdehyde content in plasma. The encouraging results of the improvement of growth performance, nutrient digestibility, antioxidant ability, and gut health parameters suggested that partially full-fatted BSFM inclusion can be suitable protein ingredients for layer chickens' diets at the starter period.

Genome shuffling to improve thermotolerance, ethanol tolerance and ethanol productivity of Saccharomyces cerevisiae.

Genome shuffling is a powerful strategy for rapid engineering of microbial strains for desirable industrial phenotypes. Here we improved the thermotolerance and ethanol tolerance of an industrial yeast strain SM-3 by genome shuffling while simultaneously enhancing the ethanol productivity. The starting population was generated by protoplast ultraviolet irradiation and then subjected for the recursive protoplast fusion. The positive colonies from the library, created by fusing the inactivated protoplasts were screened for growth at 35, 40, 45, 50 and 55 degrees C on YPD-agar plates containing different concentrations of ethanol. Characterization of all mutants and wild-type strain in the shake-flask indicated the compatibility of three phenotypes of thermotolerance, ethanol tolerance and ethanol yields enhancement. After three rounds of genome shuffling, the best performing strain, F34, which could grow on plate cultures up to 55 degrees C, was obtained. It was found capable of completely utilizing 20% (w/v) glucose at 45-48 degrees C, producing 9.95% (w/v) ethanol, and tolerating 25% (v/v) ethanol stress.