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Biology subjects

Spira, B.

Publications and source records attributed to Spira, B..

4 recordsLinked to original sources

ACKR1-expressing venous endothelial cells establish a pro-fibrotic niche in pulmonary fibrosis

Idiopathic pulmonary fibrosis (IPF) is a progressive lung disease characterized by excessive extracellular matrix deposition and irreversible architectural distortion of the lung. Fibrotic remodeling is driven by dynamic interactions among endothelial, fibroblast, epithelial and immune cells. Although pulmonary endothelial cells (ECs) are increasingly recognized as important contributors to IPF pathogenesis, the molecular and cellular events underlying endothelial dysfunction remains poorly understood. Using integrative multi-omics analyses of human IPF lungs combined with functional in vitro assays, we identify ACKR1-expressing venous endothelial cells (ACKR1+ VECs) as critical regulators of a pathogenic niche that promotes lung fibrosis. Single-cell RNA sequencing and spatial transcriptomics analyses reveal that ACKR1+ VECs exhibit a distinct pro-fibrotic and pro-inflammatory transcriptional program enriched for hypoxia responses, extracellular matrix remodeling, and immune cell recruitment. In both mouse and human fibrotic lungs, ACKR1+ VECs localize adjacent to fibroblastic foci and are surrounded by pro-fibrotic CD68+/CCR5+/SPP1+ macrophages-monocytes, suggesting a spatial organized cellular crosstalk supporting fibrotic remodeling. Consistent with these findings, in vitro co-culture assays using ACKR1+ VECs isolated from IPF lungs demonstrate that these cells drive myeloid recruitment and fibroblast activation through ACKR1 dependent mechanisms. Silencing of ACKR1 in IPF-derived VECs suppressed inflammatory and fibrotic transcriptional programs, and pharmacological inhibition of ACKR1 attenuated stromal and immune remodeling and reduced bleomycin-induced lung fibrosis in vivo. Together, these findings identify ACKR1+ VECs as key orchestrators of fibrosis progression and establish ACKR1 and the pathogenic vasculature as promising therapeutic targets for IPF. Clinical RelevanceIdiopathic pulmonary fibrosis (IPF) is a progressive and fatal lung disease with limited treatment options. We identify ACKR1-expressing venous endothelial cells as key drivers of inflammatory and fibrotic remodeling and show that pharmacologic inhibition of ACKR1 attenuates experimental lung fibrosis. These findings establish endothelial ACKR1 as a promising therapeutic target and highlight the pulmonary vasculature as a novel avenue for disease-modifying therapies in IPF.

molecular biology↗

ADP-MoA: a platform for screening antibiotic activity and their mechanism of action in Pseudomonas aeruginosa

The emergence and proliferation of multidrug-resistant bacteria pose a major threat to global public health. To address an imminent crisis, it is essential to identify and characterize new antibacterial molecules. With that in mind, we developed the ADP-MoA platform, that facilitates the discovery of new antibiotics and provides preliminary insights into their mechanisms of action. The basic idea is to simultaneously visualize antibiotic activity - growth inhibition, along with one of the three classic antibiotics mechanisms of action: DNA damage/inhibition of DNA replication, protein synthesis inhibition and cell wall damage. The platform consists of three different chromosomal fusions between the promoters of recA, ampC or armZ and the luxCDABE operon. The platform was constructed and hitherto tested in the pathogenic opportunistic bacterium Pseudomonas aeruginosa. As a proof of concept we showed that the promoter fusions were each activated by the expected antibiotics with known mechanisms of action. The armZ::luxCDABE fusion responded to antibiotics that inhibit protein synthesis (macrolides, chloramphenicol, tetracyclines and aminoglycosides), ampC::luxCDABE was induced by {beta}-lactams and recA::luxCDABE was induced by quinolones. Interestingly, ciprofloxacin induced PampC and ParmZ as well, albeit at a lower level. The ADP-MoA platform offers a readily implementable, low-cost approach with significant potential for high-throughput screening of antimicrobials against P. aeruginosa and other bacterial species.

microbiology↗

A bacterial tragedy of the commons that masks the actual frequency of mutants

The frequency of mutants in a population is central to the understanding of evolution. Mutant frequency is usually assessed by plating a bacterial culture on selective medium in which only specific rare mutants can grow, assuming that all mutant cells present on the plate are able to form colonies. Here we show an exception to this rule. Wild-type Escherichia coli cells are unable to grow with glycerol-2-phosphate (G2P) as a carbon source. In contrast, PHO-constitutive mutants can hydrolyse G2P to glycerol and form colonies on plates having G2P as their sole carbon source. However, the frequency of PHO-constitutive colonies on the selective plate is exceptionally low. Here we show that such mutations occur at a relatively high rate, but the growth of the existing mutants is inhibited due to a competition with the surrounding wild-type cells for the limited amounts of glycerol produced by the mutants. This scenario in which neither the wild-type nor the majority of the mutants are able to grow constitutes an unavoidable case of the tragedy of the commons. Evidence shows that the few mutants that do form colonies derive from micro-clusters of mutants on the selective plate. In addition, a mathematical model describes the fate of the wild-type and mutant populations on the selective plate.

genetics↗

Visible Light Plasmon Excitation of Silver Nanoparticles Against Antibiotic-Resistant Pseudomonas aeruginosa

The interaction of metallic nanoparticles with light excites a local surface plasmon resonance (LSPR). This phenomenon enables the transfer of hot electrons to substrates that release Reactive Oxygen Species (ROS). In this context, the present study was aimed at enhancing the antibacterial effect of citrate-covered silver nanoparticles (AgNPs), which already possess excellent antimicrobial properties, via LSPR excitation with visible LED against Pseudomonas aeruginosa, one of the most refractory organisms to antibiotic treatment. The Minimum Inhibitory Concentration (MIC) of AgNPs was 10 g/ml under dark conditions and 5 g/ml under light conditions. The combination of light and AgNPs led to 100% cell death after 60 minutes. Quantification of ROS via flow cytometry showed that LSPR stimulated AgNPs increased intracellular ROS concentration by 4.8-fold, suggesting that light-exposed AgNPs caused cell death via ROS production. Light exposition caused a small release of silver ions (0.4%) reaching a maximum after 6 hours. This indicates that silver ions play at most a secondary role in P. aeruginosa death. Overall, the results presented here show that LSPR generation from AgNPs by visible light enhances the antimicrobial activity of silver nanoparticles and can be an alternative for the treatment of topic infections caused by antibiotic-resistant bacteria such as P. aeruginosa.

microbiology↗