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Iacovelli, F.

Publications and source records attributed to Iacovelli, F..

4 recordsLinked to original sources

Zinc Starvation Drives Respiratory Remodeling and Metabolic Adaptations Associated with Cystic Fibrosis in Pseudomonas aeruginosa

Host nutritional immunity restricts microbial growth by altering metal availability. In patients with cystic fibrosis (CF), microorganisms inhabiting the thick airway mucus experience severe nutrient limitation, particularly zinc (Zn) restriction, which plays a critical role in limiting lung colonization. However, Pseudomonas aeruginosa, a major contributor to morbidity and mortality in CF, employs multiple strategies to overcome Zn deficiency and persist within the airways. Understanding how P. aeruginosa adapts to Zn deprivation may facilitate the development of antimicrobial approaches targeting Zn homeostasis. In this study, we characterized the physiological and transcriptional adaptations that support P. aeruginosa survival under Zn-limited conditions. Transcriptomic analysis of a znuAzrmB mutant unable to efficiently acquire Zn revealed widespread repression of pathways involved in central carbon metabolism, motility and virulence. Notably, Zn limitation promoted extensive respiratory remodeling, characterized by a shift toward anaerobic metabolism, induction of denitrification pathways, altered terminal oxidase expression, and reduced oxygen consumption. These metabolic changes correlated with decreased ATP production, altered membrane potential, and increased aminoglycoside tolerance. Furthermore, the Zn-starved mutant exhibited reduced production of quorum-sensing molecules, redox imbalance and altered oxidative stress responses. Many of these adaptations resemble those observed in P. aeruginosa isolated from CF sputum, suggesting convergence towards a common host-adapted physiological state. Collectively, these findings identify Zn starvation as a major driver of bacterial physiological remodeling in CF conditions and reveal a previously unrecognized link between Zn limitation, respiratory reprogramming, and the emergence of persistence-associated traits in P. aeruginosa. ImportanceDuring infection, the host restricts Zn availability as part of nutritional immunity, but how this influences the physiology of Pseudomonas aeruginosa remains poorly understood. Here we show that severe Zn limitation triggers a coordinated metabolic program that extends far beyond Zn acquisition, encompassing respiratory remodeling, altered energy metabolism, redox imbalance, and increased tolerance to aminoglycosides. Remarkably, many of these changes resemble transcriptional and physiological traits previously described in bacteria adapted to cystic fibrosis airways, identifying Zn limitation as a key environmental signal that contributes to chronic infection-associated phenotypes. These findings broaden our understanding of how metal availability shapes bacterial physiology and suggest that targeting Zn homeostasis may influence both bacterial persistence and antibiotic susceptibility.

microbiology↗

Investigation of Zur-regulated metal transport systems reveals an unexpected role of pyochelin in zinc homeostasis

Limiting the availability of transition metals at infection sites serves as a critical defense mechanism employed by the innate immune system to combat microbial infections. Pseudomonas aeruginosa exhibits a remarkable ability to thrive in zinc-deficient environments, which is facilitated by intricate cellular responses governed by numerous genes regulated by the zinc-responsive transcription factor Zur. Many of these genes have unknown functions, including those within the predicted PA2911-PA2914 and PA4063-PA4066 operons. A bioinformatic analysis revealed that PA2911-PA2914 comprises a TonB-dependent outer membrane receptor and an inner membrane ABC-permease responsible for importing metal-chelating molecules, whereas PA4063-PA4066 contains genes encoding a MacB transporter, likely involved in the export of large molecules. Molecular genetics and biochemical experiments, feeding assays, and intracellular metal content measurements demonstrated that PA2911-PA2914 and PA4063-PA4066 are engaged in the import and export of the pyochelin-cobalt complex, respectively. Notably, cobalt can reduce zinc demand and promote the growth of P. aeruginosa strains unable to import zinc, highlighting pyochelin-mediated cobalt import as a novel bacterial strategy to counteract zinc deficiency. These results unveil an unexpected role for pyochelin in zinc homeostasis and challenge the traditional view of this metallophore exclusively as an iron transporter.

microbiology↗

Pleiotropic effect of Lactoferrin in the prevention and treatment of COVID-19 infection: in vivo, in silico and in vitro preliminary evidences

Lactoferrin, a multifunctional cationic glycoprotein, secreted by exocrine glands and neutrophils, possesses an antiviral activity extendable to SARS-CoV-2. We performed in vitro assays proving lactoferrin antiviral activity through direct attachment to both virus and cell surface components. This activity varied according to concentration (100/500g/ml), multiplicity of infection (0.1/0.01) and cell type (Vero E6/Caco-2 cells). Interestingly, the in silico results strongly supported the hypothesis of a direct recognition between the lactoferrin and the Spike S glycoprotein, thus hindering the viral entry into the cells. Hence, we conducted a clinical trial to investigate effect and tolerability of a liposomal lactoferrin formulation as a supplementary nutraceutical agent in mild-to-moderate and asymptomatic COVID-19 patients. A total of 92 mild-to-moderate (67/92) and asymptomatic (25/92) COVID-19 patients were recruited and divided in 3 groups according to the administered regimen. Thirty-two patients, 14 hospitalised and 18 in home-based insolation received oral and intranasal liposomal bovine lactoferrin (bLf), 32 hospitalised patients were treated with standard of care treatment (hydroxychloroquine, azitromicin and lopinavir/darunavir), and 28, in home-based isolation, did not take any medication. Furthermore, 32 COVID-19 negative, not-treated, healthy subjects were added as a control group for ancillary analysis. bLf-supplemented COVID-19 patients obtained an earlier and significant (p < 0,0001.) median rRT-PCR SARS-COV-2 RNA negative conversion than standard of care-treated and non-treated COVID-19 patients (14.25 vs 27.13 vs 32.61 days, respectively). In addition, bLf-supplemented COVID-19 patients showed significant fast clinical symptoms recovery than standard of care-treated and non-treated COVID-19 patients. Moreover, in bLf-supplemented patients, a significant decrease of either serum ferritin or IL-6 levels or host iron overload, all parameters characterizing inflammatory processes, were observed. Serum D-dimers was also found significantly decreased following bLf supplement. No adverse events were reported. These in vitro and in vivo observations led us to speculate a potential and safe supplementary role of Blf in the management of mild-to-moderate and asymptomatic COVID-19 patients.

microbiology↗

Frataxin deficiency induces lipid accumulation and affects thermogenesis in brown adipose tissue

Decreased expression of the mitochondrial protein frataxin (FXN) causes Friedreichs ataxia (FRDA). FRDA is a neurodegenerative disease also characterized by systemic metabolic alterations that increase the risk of developing type 2 diabetes thus aggravating FRDA prognosis. Brown adipose tissue (BAT) is a mitochondria-enriched and anti-diabetic tissue that, in addition to its thermoregulatory role, turns excess energy into heat to maintain energy balance. Here we report that the FXN knock-in/knock-out (KIKO) mouse shows reduced energy expenditure and VO2, hyperlipidemia, decreased insulin sensitivity and enhanced circulating levels of leptin, recapitulating diabetes-like signatures. FXN deficiency leads to alteration of mitochondrial structure and oxygen consumption, decreased lipolysis and lipid accumulation in BAT. Transcriptomic data highlighted a blunted thermogenesis response, as several biological processes related to thermogenesis (e.g. response to temperature stimuli, mitochondrial gene transcription, triglyceride metabolism, adipogenesis) resulted affected in BAT of KIKO mice upon cold exposure. Decreased adaptation to cool temperature in association with limited PKA-mediated lipolysis and downregulation of the expression of the genes controlling mitochondrial metabolism and lipid catabolism were observed in KIKO mice. T37i brown adipocytes and primary adipocytes with FXN deficiency showed reduced thermogenesis and adipogenesis markers respectively recapitulating the molecular signatures detected in KIKO mice.\n\nCollectively our data point to BAT dysfunction in FRDA and suggest BAT as a promising target to overcome metabolic complications in FRDA.

pathology↗