Cortez, R. | Search bioRxiv

bioRxiv2025

Kasugamycin inhibits melanoma lung metastasis and regulates CHI3L1-driven M2-like tumor-associated macrophage differentiation

CHI3L1, a chitinase-like protein, is a potent immune modulator involved in various diseases, including lung cancer. While recent studies have demonstrated that kasugamycin (KSM) is a pan-chitinase inhibitor with strong anti-fibrotic activity, its effects on specific chitinase-like proteins remain undefined. This study shows that KSM effectively abrogates CHI3L1-stimulated cellular signaling and bioactivities. In a B16/F10 melanoma lung metastasis model, where CHI3L1 plays a critical role, KSM treatment significantly reduced melanoma lung metastasis dose-dependently. The anti-tumor effect of KSM was found to be CHI3L1-specific, as CHI3L1 overexpression enhanced melanoma lung colony formation, which was effectively blocked by KSM. In melanoma-challenged lungs, KSM treatment significantly reduced the elevation of M2 macrophages expressing CD206, CD163, and PD-L1. In studies using human monocytic THP-1 cells, CHI3L1 promoted M2 macrophage differentiation, which KSM significantly suppressed. Bulk RNA sequencing of differentiated macrophages revealed that CHI3L1 highly induced the expression of epidermal growth factor receptor (EGFR), and this induction was counter-regulated by KSM, and CHI3L1-driven M2 macrophage activation was reduced with EGFR blocker treatment. These findings reveal a novel anti-tumor mechanism of KSM, which inhibits M2-like tumor-associated macrophage differentiation, potentially through the CHI3L1-EGFR axis.

immunology↗

bioRxiv2024

Co-occurrence of Direct and Indirect Extracellular Electron Transfer Mechanisms during Electroactive Respiration in a Dissimilatory Sulfate Reducing Bacterium

Extracellular electron transfer (EET) propels microbial fuel cell (MFC) technology and contributes to the mobility of redox active minerals and microbial syntrophy in nature. Sulfate-reducing bacteria (SRB), especially the genus Desulfovibrio corrode metal electrodes but are of interest for sulfate-containing MFCs providing wastewater treatment. Although extensive studies on SRB-mediated metal electrode corrosion have been done, there remain knowledge gaps on SRB EET to electrodes. We aimed to determine SRB EET mechanisms towards improving SRB performance in MFC wastewater treatment. Our MFCs with Desulfovibrio vulgaris Hildenborough (DvH), a model SRB, indicated that DvH can harvest and send electrons to the carbon cloth electrode. Electricity production with a maximum power density of [~]0.074 W/m2 was observed when the ratio of lactate (electron and carbon donor) to sulfate (electron acceptor) was 60:20 and 0:10 in the anodic and cathodic chamber, respectively. Patterns in current production compared to variations of electron donor/acceptor ratios in the anode and cathode suggested that attachment of DvH to the electrode and biofilm density were critical for effective electricity generation. Analysis of DvH biofilms at different conditions (planktonic dissimilatory sulfate reduction respiration vs. electroactive respiration) by electron microscopy indicated DvH utilized filaments that resemble nano-pili to attach on electrodes and facilitate EET from cell-to-cell and to the electrode. Proteomics profiling of electroactive respiration proteins indicated DvH adapted to electroactive respiration by presenting more pili-, flagellar-related proteins and histidine kinases on electrodes. To investigate the role of pili and biofilm, we grew two DvH mutants in MFCs under the same conditions. The mutant with a deletion of the major pilus-producing gene yielded less voltage and far less attachment to the electrode, suggesting the importance of pili in EET. The mutant with a deficiency in biofilm formation, however, did not eliminate current production indicating the existence of indirect EET. Untargeted metabolomics profiling showed flavin-based metabolites, potential electron shuttles, were dysregulated between respiration modes. This work revealed the metabolic flexibility of DvH to thrive in less than ideal conditions with solid surfaces as both an electron acceptor (growth on anode) and donor (growth on cathode) by using a combination of direct and indirect EET mechanisms. Understanding DvH EET mechanism could enhance the application of DvH in MFCs treating wastewater. ImportanceWe explored the application of Desulfovibrio vulgaris Hildenborough in microbial fuel cells (MFC) and investigated its potential extracellular electron transfer (EET) mechanism. We also conducted untargeted proteomics and metabolomics profiling, offering insights into how DvH adapts metabolically to different electron donors and acceptors. An understanding of the EET mechanism and metabolic flexibility of DvH holds promise for future uses including bioremediation or enhancing efficacy in MFCs for wastewater treatment applications.

microbiology↗