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Libisch B. N-Alkane Assimilation by Pseudomonas aeruginosa and Its Interactions with Virulence and Antibiotic Resistance. Antibiotics (Basel) 2024; 13:1028. [PMID: 39596723 PMCID: PMC11591199 DOI: 10.3390/antibiotics13111028] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/15/2024] [Revised: 10/24/2024] [Accepted: 10/28/2024] [Indexed: 11/29/2024] Open
Abstract
Pseudomonas aeruginosa strains with potential for degrading n-alkanes are frequently cultured from hydrocarbon-contaminated sites. The initial hydroxylation step of long-chain n-alkanes is mediated by the chromosomally encoded AlkB1 and AlkB2 alkane hydroxylases. The acquisition of an additional P. putida GPo1-like alkane hydroxylase gene cluster can extend the substrate range assimilated by P. aeruginosa to
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Affiliation(s)
- Balázs Libisch
- Institute of Genetics and Biotechnology, Hungarian University of Agriculture and Life Sciences, H-2100 Gödöllő, Hungary
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2
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Nouh HS, El-Zawawy NA, Halawa M, Shalamesh EM, Ali SS, Korbecka-Glinka G, Shala AY, El-Sapagh S. Endophytic Penicillium oxalicum AUMC 14898 from Opuntia ficus-indica: A Novel Source of Tannic Acid Inhibiting Virulence and Quorum Sensing of Extensively Drug-Resistant Pseudomonas aeruginosa. Int J Mol Sci 2024; 25:11115. [PMID: 39456896 PMCID: PMC11507641 DOI: 10.3390/ijms252011115] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/16/2024] [Revised: 10/08/2024] [Accepted: 10/11/2024] [Indexed: 10/28/2024] Open
Abstract
Pseudomonas aeruginosa is a harmful pathogen that causes a variety of acute and chronic infections through quorum sensing (QS) mechanisms. The increasing resistance of this bacterium to numerous antibiotics has created a demand for new medications that specifically target QS. Endophytes can be the source of compounds with antibacterial properties. This research is the first to examine tannic acid (TA) produced by endophytic fungus as a potential biotherapeutic agent. A novel endophytic fungal isolate identified as Penicillium oxalicum was derived from the cladodes of Opuntia ficus-indica (L.). The species identification for this isolate was confirmed through sequencing of the internal transcribed spacer region. The metabolites from the culture of this isolate were extracted using ethyl acetate, then separated and characterized using chromatographic methods. This led to the acquisition of TA, a compound that shows strong anti-QS and excellent antibacterial effects against extensively drug-resistant P. aeruginosa strains. Furthermore, it was shown that treating P. aeruginosa with the obtained TA reduced the secretion of virulence factors controlled by QS in a dose-dependent manner, indicating that TA inhibited the QS characteristics of P. aeruginosa. Simultaneously, TA significantly inhibited the expression of genes associated with QS, including rhlR/I, lasR/I, and pqsR. In addition, in silico virtual molecular docking showed that TA could efficiently bind to QS receptor proteins. Our results showed that P. oxalicum could be a new source of TA for the treatment of infections caused by extensively drug-resistant P. aeruginosa.
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Affiliation(s)
- Hoda S. Nouh
- Botany and Microbiology Department, Faculty of Science, Tanta University, Tanta 31511, Egypt; (N.A.E.-Z.); (M.H.); (E.M.S.); (S.S.A.); (S.E.-S.)
| | - Nessma A. El-Zawawy
- Botany and Microbiology Department, Faculty of Science, Tanta University, Tanta 31511, Egypt; (N.A.E.-Z.); (M.H.); (E.M.S.); (S.S.A.); (S.E.-S.)
| | - Mohamed Halawa
- Botany and Microbiology Department, Faculty of Science, Tanta University, Tanta 31511, Egypt; (N.A.E.-Z.); (M.H.); (E.M.S.); (S.S.A.); (S.E.-S.)
| | - Ebrahim M. Shalamesh
- Botany and Microbiology Department, Faculty of Science, Tanta University, Tanta 31511, Egypt; (N.A.E.-Z.); (M.H.); (E.M.S.); (S.S.A.); (S.E.-S.)
| | - Sameh Samir Ali
- Botany and Microbiology Department, Faculty of Science, Tanta University, Tanta 31511, Egypt; (N.A.E.-Z.); (M.H.); (E.M.S.); (S.S.A.); (S.E.-S.)
| | - Grażyna Korbecka-Glinka
- Department of Biotechnology and Plant Breeding, Institute of Soil Science and Plant Cultivation—State Research Institute, 24-100 Puławy, Poland
| | - Awad Y. Shala
- Medicinal and Aromatic Plants Research Department, Horticulture Research Institute, Agricultural Research Center, Giza 12619, Egypt;
| | - Shimaa El-Sapagh
- Botany and Microbiology Department, Faculty of Science, Tanta University, Tanta 31511, Egypt; (N.A.E.-Z.); (M.H.); (E.M.S.); (S.S.A.); (S.E.-S.)
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Ostovar G, Boedicker JQ. Phenotypic memory in quorum sensing. PLoS Comput Biol 2024; 20:e1011696. [PMID: 38976753 PMCID: PMC11257393 DOI: 10.1371/journal.pcbi.1011696] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/07/2023] [Revised: 07/18/2024] [Accepted: 06/19/2024] [Indexed: 07/10/2024] Open
Abstract
Quorum sensing (QS) is a regulatory mechanism used by bacteria to coordinate group behavior in response to high cell densities. During QS, cells monitor the concentration of external signals, known as autoinducers, as a proxy for cell density. QS often involves positive feedback loops, leading to the upregulation of genes associated with QS signal production and detection. This results in distinct steady-state concentrations of QS-related molecules in QS-ON and QS-OFF states. Due to the slow decay rates of biomolecules such as proteins, even after removal of the initial stimuli, cells can retain elevated levels of QS-associated biomolecules for extended periods of time. This persistence of biomolecules after the removal of the initial stimuli has the potential to impact the response to future stimuli, indicating a memory of past exposure. This phenomenon, which is a consequence of the carry-over of biomolecules rather than genetic inheritance, is known as "phenotypic" memory. This theoretical study aims to investigate the presence of phenotypic memory in QS and the conditions that influence this memory. Numerical simulations based on ordinary differential equations and analytical modeling were used to study gene expression in response to sudden changes in cell density and extracellular signal concentrations. The model examined the effect of various cellular parameters on the strength of QS memory and the impact on gene regulatory dynamics. The findings revealed that QS memory has a transient effect on the expression of QS-responsive genes. These consequences of QS memory depend strongly on how cell density was perturbed, as well as various cellular parameters, including the Fold Change in the expression of QS-regulated genes, the autoinducer synthesis rate, the autoinducer threshold required for activation, and the cell growth rate.
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Affiliation(s)
- Ghazaleh Ostovar
- Department of Physics and Astronomy, University of Southern California, Los Angeles, California, United States of America
| | - James Q. Boedicker
- Department of Physics and Astronomy, University of Southern California, Los Angeles, California, United States of America
- Department of Biological Sciences, University of Southern California, Los Angeles, California, United States of America
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Xu Q, Kang D, Meyer MD, Pennington CL, Gopal C, Schertzer JW, Kirienko NV. Cytotoxic rhamnolipid micelles drive acute virulence in Pseudomonas aeruginosa. Infect Immun 2024; 92:e0040723. [PMID: 38391248 PMCID: PMC10929412 DOI: 10.1128/iai.00407-23] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/08/2023] [Accepted: 02/07/2024] [Indexed: 02/24/2024] Open
Abstract
Pseudomonas aeruginosa is an opportunistic human pathogen that has developed multi- or even pan-drug resistance toward most frontline and last resort antibiotics, leading to increasing frequency of infections and deaths among hospitalized patients, especially those with compromised immune systems. Further complicating treatment, P. aeruginosa produces numerous virulence factors that contribute to host tissue damage and immune evasion, promoting bacterial colonization and pathogenesis. In this study, we demonstrate the importance of rhamnolipid production in host-pathogen interactions. Secreted rhamnolipids form micelles that exhibited highly acute toxicity toward murine macrophages, rupturing the plasma membrane and causing organellar membrane damage within minutes of exposure. While rhamnolipid micelles (RMs) were particularly toxic to macrophages, they also caused membrane damage in human lung epithelial cells, red blood cells, Gram-positive bacteria, and even noncellular models like giant plasma membrane vesicles. Most importantly, rhamnolipid production strongly correlated with P. aeruginosa virulence against murine macrophages in various panels of clinical isolates. Altogether, our findings suggest that rhamnolipid micelles are highly cytotoxic virulence factors that drive acute cellular damage and immune evasion during P. aeruginosa infections.
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Affiliation(s)
- Qi Xu
- Department of BioSciences, Rice University, Houston, Texas, USA
- Department of Bioengineering, Rice University, Houston, Texas, USA
| | - Donghoon Kang
- Department of BioSciences, Rice University, Houston, Texas, USA
| | - Matthew D. Meyer
- Shared Equipment Authority, Rice University, Houston, Texas, USA
| | | | - Citrupa Gopal
- Department of Biological Sciences, Binghamton University, Binghamton, New York, USA
| | - Jeffrey W. Schertzer
- Department of Biological Sciences, Binghamton University, Binghamton, New York, USA
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5
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Xu Q, Kang D, Meyer MD, Pennington CL, Gopal C, Schertzer JW, Kirienko NV. Cytotoxic rhamnolipid micelles drive acute virulence in Pseudomonas aeruginosa. BIORXIV : THE PREPRINT SERVER FOR BIOLOGY 2023:2023.10.13.562257. [PMID: 37873290 PMCID: PMC10592815 DOI: 10.1101/2023.10.13.562257] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 10/25/2023]
Abstract
Pseudomonas aeruginosa is an opportunistic human pathogen that has developed multi- or even pan-drug resistance towards most frontline and last resort antibiotics, leading to increasing infections and deaths among hospitalized patients, especially those with compromised immune systems. Further complicating treatment, P. aeruginosa produces numerous virulence factors that contribute to host tissue damage and immune evasion, promoting bacterial colonization and pathogenesis. In this study, we demonstrate the importance of rhamnolipid production in host-pathogen interactions. Secreted rhamnolipids form micelles that exhibited highly acute toxicity towards murine macrophages, rupturing the plasma membrane and causing organellar membrane damage within minutes of exposure. While rhamnolipid micelles (RMs) were particularly toxic to macrophages, they also caused membrane damage in human lung epithelial cells, red blood cells, Gram-positive bacteria, and even non-cellular models like giant plasma membrane vesicles. Most importantly, rhamnolipid production strongly correlated to P. aeruginosa virulence against murine macrophages in various panels of clinical isolates. Altogether, our findings suggest that rhamnolipid micelles are highly cytotoxic virulence factors that drive acute cellular damage and immune evasion during P. aeruginosa infections.
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Affiliation(s)
- Qi Xu
- Department of BioSciences, Rice University, Houston, Texas, USA
- Department of Bioengineering, Rice University, Houston, Texas, USA
| | - Donghoon Kang
- Department of BioSciences, Rice University, Houston, Texas, USA
| | - Matthew D. Meyer
- Shared Equipment Authority, Rice University, Houston, Texas, USA
| | | | - Citrupa Gopal
- Department of Biological Sciences, Binghamton University, Binghamton, New York, USA
| | - Jeffrey W. Schertzer
- Department of Biological Sciences, Binghamton University, Binghamton, New York, USA
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Sass G, Groleau MC, Déziel E, Stevens DA. Simple method for quantification of anionic biosurfactants in aqueous solutions. Front Bioeng Biotechnol 2023; 11:1253652. [PMID: 37885452 PMCID: PMC10598384 DOI: 10.3389/fbioe.2023.1253652] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/05/2023] [Accepted: 09/13/2023] [Indexed: 10/28/2023] Open
Abstract
Biosurfactants are microbial products that have applications as cleaning agents, emulsifiers, and dispersants. Detection and quantification of biosurfactants can be done by various methods, including colorimetric tests, high performance liquid chromatography (HPLC) coupled to several types of detectors, and tests that take advantage of biosurfactants reducing surface tension of aqueous liquids, allowing for spreading and droplet formation of oils. We present a new and simple method for quantifying biosurfactants by their ability, on paper, to reduce surface tension of aqueous solutions, causing droplet dispersion on an oiled surface in correlation with biosurfactant content. We validated this method with rhamnolipids, surfactin, sophorolipids, and ananatoside B; all are anionic microbial surfactants. Linear ranges for quantification in aqueous solutions for all tested biosurfactants were between 10 and 500 µM. Our method showed time-dependent biosurfactant accumulation in cultures of Pseudomonas aeruginosa strains PA14 and PAO1, and Burkholderia thailandensis E264. Mutants in genes responsible for surfactant production showed negligible activity on oiled paper. In summary, our simple assay provides the opportunity to quantify biosurfactant contents of aqueous solutions, for a diversity of surfactants, by means readily available in any laboratory.
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Affiliation(s)
- Gabriele Sass
- California Institute for Medical Research, San Jose, CA, United States
| | - Marie-Christine Groleau
- Institut National de la Recherche Scientific-Centre Armand-Frappier Santé Biotechnologie, Laval, QC, Canada
| | - Eric Déziel
- Institut National de la Recherche Scientific-Centre Armand-Frappier Santé Biotechnologie, Laval, QC, Canada
| | - David A. Stevens
- California Institute for Medical Research, San Jose, CA, United States
- Division of Infectious Diseases and Geographic Medicine, Department of Medicine, Stanford University School of Medicine, Stanford, CA, United States
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7
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Abd El-Aleam RH, Sayed AM, Taha MN, George RF, Georgey HH, Abdel-Rahman HM. Design and synthesis of novel benzimidazole derivatives as potential Pseudomonas aeruginosa anti-biofilm agents inhibiting LasR: Evidence from comprehensive molecular dynamics simulation and in vitro investigation. Eur J Med Chem 2022; 241:114629. [DOI: 10.1016/j.ejmech.2022.114629] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/02/2022] [Revised: 07/10/2022] [Accepted: 07/21/2022] [Indexed: 11/04/2022]
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Tuon FF, Dantas LR, Suss PH, Tasca Ribeiro VS. Pathogenesis of the Pseudomonas aeruginosa Biofilm: A Review. Pathogens 2022; 11:pathogens11030300. [PMID: 35335624 PMCID: PMC8950561 DOI: 10.3390/pathogens11030300] [Citation(s) in RCA: 163] [Impact Index Per Article: 54.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/22/2021] [Revised: 02/08/2022] [Accepted: 02/24/2022] [Indexed: 01/21/2023] Open
Abstract
Pseudomonas aeruginosa is associated with several human infections, mainly related to healthcare services. In the hospital, it is associated with resistance to several antibiotics, which poses a great challenge to therapy. However, one of the biggest challenges in treating P. aeruginosa infections is that related to biofilms. The complex structure of the P. aeruginosa biofilm contributes an additional factor to the pathogenicity of this microorganism, leading to therapeutic failure, in addition to escape from the immune system, and generating chronic infections that are difficult to eradicate. In this review, we address several molecular aspects of the pathogenicity of P. aeruginosa biofilms.
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Lichtenberg M, Jakobsen TH, Kühl M, Kolpen M, Jensen PØ, Bjarnsholt T. OUP accepted manuscript. FEMS Microbiol Rev 2022; 46:6574409. [PMID: 35472245 PMCID: PMC9438473 DOI: 10.1093/femsre/fuac018] [Citation(s) in RCA: 19] [Impact Index Per Article: 6.3] [Reference Citation Analysis] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/30/2021] [Revised: 04/04/2022] [Accepted: 04/24/2022] [Indexed: 11/13/2022] Open
Affiliation(s)
- Mads Lichtenberg
- Costerton Biofilm Center, Department of Immunology and Microbiology, University of Copenhagen, Blegdamsvej 3B, 2200, København, Denmark
| | - Tim Holm Jakobsen
- Costerton Biofilm Center, Department of Immunology and Microbiology, University of Copenhagen, Blegdamsvej 3B, 2200, København, Denmark
| | - Michael Kühl
- Marine Biological Section, Department of Biology, University of Copenhagen, Strandpromenaden 5, 3000 Helsingør, Denmark
| | - Mette Kolpen
- Department of Clinical Microbiology, Copenhagen University Hospital, Ole Maaløes vej 26, 2200, København, Denmark
| | - Peter Østrup Jensen
- Costerton Biofilm Center, Department of Immunology and Microbiology, University of Copenhagen, Blegdamsvej 3B, 2200, København, Denmark
- Department of Clinical Microbiology, Copenhagen University Hospital, Ole Maaløes vej 26, 2200, København, Denmark
| | - Thomas Bjarnsholt
- Corresponding author: Costerton Biofilm Center, Department of Immunology and Microbiology, University of Copenhagen, Blegdamsvej 3B, 2200, København, Denmark. Tel: +45 20659888; E-mail:
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10
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Franklin T, Wu Y, Lang J, Li S, Yang R. Design of Polymeric Thin Films to Direct Microbial Biofilm Growth, Virulence, and Metabolism. Biomacromolecules 2021; 22:4933-4944. [PMID: 34694768 DOI: 10.1021/acs.biomac.1c00731] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/30/2022]
Abstract
Biofilms are ubiquitous in nature, yet strategies to direct biofilm behavior without genetic manipulation are limited. Due to the small selection of materials that have been used to successfully grow biofilms, the availability of functional materials that are able to support growth and program microbial functions remains a critical bottleneck in the design and deployment of functional yet safe microbes. Here, we report the design of insoluble pyridine-rich polymer surfaces synthesized using initiated chemical vapor deposition, which led to modulated biofilm growth and virulence in Pseudomonas aeruginosa (PAO1). A variety of extracellular virulence factors exhibited decreased production in response to the functional polymer, most significantly biomolecules also associated with iron acquisition, validating the material design strategy reported here. This report signifies a rich potential for materials-based strategies to direct the behavior of naturally occurring biofilms, which complement the existing genetic engineering toolkits in advancing microbiology, translational medicine, and biomanufacturing.
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Affiliation(s)
- Trevor Franklin
- Robert F. Smith School of Chemical & Biomolecular Engineering, Cornell University, 120, Olin Hall, Ithaca, New York 14853, United States
| | - Yinan Wu
- Robert F. Smith School of Chemical & Biomolecular Engineering, Cornell University, 120, Olin Hall, Ithaca, New York 14853, United States
| | - Jiayan Lang
- Robert F. Smith School of Chemical & Biomolecular Engineering, Cornell University, 120, Olin Hall, Ithaca, New York 14853, United States
| | - Sijin Li
- Robert F. Smith School of Chemical & Biomolecular Engineering, Cornell University, 120, Olin Hall, Ithaca, New York 14853, United States
| | - Rong Yang
- Robert F. Smith School of Chemical & Biomolecular Engineering, Cornell University, 120, Olin Hall, Ithaca, New York 14853, United States
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Singh S, Bhatia S. Quorum Sensing Inhibitors: Curbing Pathogenic Infections through Inhibition of Bacterial Communication. IRANIAN JOURNAL OF PHARMACEUTICAL RESEARCH : IJPR 2021; 20:486-514. [PMID: 34567177 PMCID: PMC8457738 DOI: 10.22037/ijpr.2020.113470.14318] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Indexed: 11/18/2022]
Abstract
Currently, most of the developed and developing countries are facing the problem of infectious diseases. The genius way of an exaggerated application of antibiotics led the infectious agents to respond by bringing a regime of persisters to resist antibiotics attacks prolonging their survival. Persisters have the dexterity to communicate among themself using signal molecules via the process of Quorum Sensing (QS), which regulates virulence gene expression and biofilms formation, making them more vulnerable to antibiotic attack. Our review aims at the different approaches applied in the ordeal to solve the riddle for QS inhibitors. QS inhibitors, their origin, structures and key interactions for QS inhibitory activity have been summarized. Solicitation of a potent QS inhibitor molecule would be beneficial, giving new life to the simplest antibiotics in adjuvant therapy.
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Affiliation(s)
- Shaminder Singh
- Regional Centre for Biotechnology, NCR Biotech Science Cluster, 3 Milestone, Faridabad-Gurugram Expressway, Faridabad - 121 001, Haryana, India
| | - Sonam Bhatia
- Department of Pharmaceutical Science, SHALOM Institute of Health and Allied Sciences, Sam Higginbottom University of Agriculture, Technology and Sciences (SHUATS), Naini-211007, Prayagraj, Uttar Pradesh, India
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12
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Chadha J, Harjai K, Chhibber S. Revisiting the virulence hallmarks of Pseudomonas aeruginosa: a chronicle through the perspective of quorum sensing. Environ Microbiol 2021; 24:2630-2656. [PMID: 34559444 DOI: 10.1111/1462-2920.15784] [Citation(s) in RCA: 91] [Impact Index Per Article: 22.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/03/2021] [Revised: 09/16/2021] [Accepted: 09/17/2021] [Indexed: 12/15/2022]
Abstract
Pseudomonas aeruginosa is an opportunistic pathogen and the leading cause of mortality among immunocompromised patients in clinical setups. The hallmarks of virulence in P. aeruginosa encompass six biologically competent attributes that cumulatively drive disease progression in a multistep manner. These multifaceted hallmarks lay the principal foundation for rationalizing the complexities of pseudomonal infections. They include factors for host colonization and bacterial motility, biofilm formation, production of destructive enzymes, toxic secondary metabolites, iron-chelating siderophores and toxins. This arsenal of virulence hallmarks is fostered and stringently regulated by the bacterial signalling system called quorum sensing (QS). The central regulatory functions of QS in controlling the timely expression of these virulence hallmarks for adaptation and survival drive the disease outcome. This review describes the intricate mechanisms of QS in P. aeruginosa and its role in shaping bacterial responses, boosting bacterial fitness. We summarize the virulence hallmarks of P. aeruginosa, relating them with the QS circuitry in clinical infections. We also examine the role of QS in the development of drug resistance and propose a novel antivirulence therapy to combat P. aeruginosa infections. This can prove to be a next-generation therapy that may eventually become refractory to the use of conventional antimicrobial treatments.
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Affiliation(s)
- Jatin Chadha
- Department of Microbiology, Panjab University, Chandigarh, India
| | - Kusum Harjai
- Department of Microbiology, Panjab University, Chandigarh, India
| | - Sanjay Chhibber
- Department of Microbiology, Panjab University, Chandigarh, India
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Morin CD, Déziel E, Gauthier J, Levesque RC, Lau GW. An Organ System-Based Synopsis of Pseudomonas aeruginosa Virulence. Virulence 2021; 12:1469-1507. [PMID: 34180343 PMCID: PMC8237970 DOI: 10.1080/21505594.2021.1926408] [Citation(s) in RCA: 33] [Impact Index Per Article: 8.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/27/2022] Open
Abstract
Driven in part by its metabolic versatility, high intrinsic antibiotic resistance, and a large repertoire of virulence factors, Pseudomonas aeruginosa is expertly adapted to thrive in a wide variety of environments, and in the process, making it a notorious opportunistic pathogen. Apart from the extensively studied chronic infection in the lungs of people with cystic fibrosis (CF), P. aeruginosa also causes multiple serious infections encompassing essentially all organs of the human body, among others, lung infection in patients with chronic obstructive pulmonary disease, primary ciliary dyskinesia and ventilator-associated pneumonia; bacteremia and sepsis; soft tissue infection in burns, open wounds and postsurgery patients; urinary tract infection; diabetic foot ulcers; chronic suppurative otitis media and otitis externa; and keratitis associated with extended contact lens use. Although well characterized in the context of CF, pathogenic processes mediated by various P. aeruginosa virulence factors in other organ systems remain poorly understood. In this review, we use an organ system-based approach to provide a synopsis of disease mechanisms exerted by P. aeruginosa virulence determinants that contribute to its success as a versatile pathogen.
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Affiliation(s)
- Charles D Morin
- Centre Armand-Frappier Santé Biotechnologie, Institut National De La Recherche Scientifique (INRS), Laval, Quebec, Canada
| | - Eric Déziel
- Centre Armand-Frappier Santé Biotechnologie, Institut National De La Recherche Scientifique (INRS), Laval, Quebec, Canada
| | - Jeff Gauthier
- Département De Microbiologie-infectiologie Et Immunologie, Institut De Biologie Intégrative Et Des Systèmes (IBIS), Université Laval, Québec City, Quebec, Canada
| | - Roger C Levesque
- Département De Microbiologie-infectiologie Et Immunologie, Institut De Biologie Intégrative Et Des Systèmes (IBIS), Université Laval, Québec City, Quebec, Canada
| | - Gee W Lau
- Department of Pathobiology, University of Illinois at Urbana-Champaign, Urbana, IL, US
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14
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Qi X, Zhang L, Xu J, Tao Z, Wang X, Qiu Y, Pan T, Liu Z, Qu H, Tan R, Liu J. Association of Increased Circulating Acetic Acid With Poor Survival in Pseudomonas aeruginosa Ventilator-Associated Pneumonia Patients. Front Cell Infect Microbiol 2021; 11:669409. [PMID: 33996639 PMCID: PMC8117141 DOI: 10.3389/fcimb.2021.669409] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/18/2021] [Accepted: 04/14/2021] [Indexed: 11/13/2022] Open
Abstract
Background We previously found that microbial disruption in Pseudomonas aeruginosa ventilator-associated pneumonia (PA-VAP) patients are long-lasting. Long-term microbial dysbiosis may lead to changes in metabolites. Short-chain fatty acids (SCFAs) are microbial fermentation products and show beneficial effects in patients with pneumonia. In this study, we aimed to explore the association between circulating SCFA levels and clinical outcomes in patients with PA-VAP. Methods In this study, we analyzed SCFAs in the serum of 49 patients with PA-VAP by gas chromatography-mass spectrometry analysis. Twenty of these patients died, and 29 survived. The correlation between serum SCFAs and patient survival and immune parameters was analyzed. Results We developed a partial least squares discriminant analysis (PLS-DA) model to examine differential SCFAs in 49 patients with PA-VAP. Among the seven SCFAs, only acetic acid was increased in non-survivors (P = 0.031, VIP > 1). Furthermore, high levels of acetic acid (>1.96ug/ml) showed increased 90-day mortality compared to low levels of acetic acid (<1.96ug/ml) in Kaplan-Meier survival analyses (P = 0.027). Increased acetic acid also correlated with reduced circulating lymphocyte and monocyte counts. Conclusion Our study showed that increased circulating acetic acid is associated with 90-day mortality in PA-VAP patients. The decrease in lymphocytes and monocytes might be affected by acetic acid and involved in the poor prognosis.
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Affiliation(s)
- Xiaoling Qi
- Department of Critical Care Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
| | - Li Zhang
- Department of Critical Care Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
| | - Jing Xu
- Department of Critical Care Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
| | - Zheying Tao
- Department of Critical Care Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
| | - Xiaoli Wang
- Department of Critical Care Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
| | - Yuzhen Qiu
- Department of Critical Care Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
| | - Tingting Pan
- Department of Critical Care Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
| | - Zhaojun Liu
- Department of Critical Care Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
| | - Hongping Qu
- Department of Critical Care Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
| | - Ruoming Tan
- Department of Critical Care Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
| | - Jialin Liu
- Department of Critical Care Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
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15
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Cendra MDM, Torrents E. Pseudomonas aeruginosa biofilms and their partners in crime. Biotechnol Adv 2021; 49:107734. [PMID: 33785375 DOI: 10.1016/j.biotechadv.2021.107734] [Citation(s) in RCA: 86] [Impact Index Per Article: 21.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/26/2020] [Revised: 03/15/2021] [Accepted: 03/18/2021] [Indexed: 12/24/2022]
Abstract
Pseudomonas aeruginosa biofilms and the capacity of the bacterium to coexist and interact with a broad range of microorganisms have a substantial clinical impact. This review focuses on the main traits of P. aeruginosa biofilms, such as the structural composition and regulatory networks involved, placing particular emphasis on the clinical challenges they represent in terms of antimicrobial susceptibility and biofilm infection clearance. Furthermore, the ability of P. aeruginosa to grow together with other microorganisms is a significant pathogenic attribute with clinical relevance; hence, the main microbial interactions of Pseudomonas are especially highlighted and detailed throughout this review. This article also explores the infections caused by single and polymicrobial biofilms of P. aeruginosa and the current models used to recreate them under laboratory conditions. Finally, the antimicrobial and antibiofilm strategies developed against P. aeruginosa mono and multispecies biofilms are detailed at the end of this review.
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Affiliation(s)
- Maria Del Mar Cendra
- Bacterial Infections and Antimicrobial therapies Group, Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology, Baldiri Reixac 15-21, 08028 Barcelona, Spain.
| | - Eduard Torrents
- Bacterial Infections and Antimicrobial therapies Group, Institute for Bioengineering of Catalonia (IBEC), The Barcelona Institute of Science and Technology, Baldiri Reixac 15-21, 08028 Barcelona, Spain; Microbiology Section, Department of Genetics, Microbiology and Statistics, Faculty of Biology, University of Barcelona, 643 Diagonal Ave., 08028 Barcelona, Spain.
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16
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Jurado-Martín I, Sainz-Mejías M, McClean S. Pseudomonas aeruginosa: An Audacious Pathogen with an Adaptable Arsenal of Virulence Factors. Int J Mol Sci 2021; 22:3128. [PMID: 33803907 PMCID: PMC8003266 DOI: 10.3390/ijms22063128] [Citation(s) in RCA: 328] [Impact Index Per Article: 82.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/28/2021] [Revised: 03/16/2021] [Accepted: 03/16/2021] [Indexed: 12/13/2022] Open
Abstract
Pseudomonas aeruginosa is a dominant pathogen in people with cystic fibrosis (CF) contributing to morbidity and mortality. Its tremendous ability to adapt greatly facilitates its capacity to cause chronic infections. The adaptability and flexibility of the pathogen are afforded by the extensive number of virulence factors it has at its disposal, providing P. aeruginosa with the facility to tailor its response against the different stressors in the environment. A deep understanding of these virulence mechanisms is crucial for the design of therapeutic strategies and vaccines against this multi-resistant pathogen. Therefore, this review describes the main virulence factors of P. aeruginosa and the adaptations it undergoes to persist in hostile environments such as the CF respiratory tract. The very large P. aeruginosa genome (5 to 7 MB) contributes considerably to its adaptive capacity; consequently, genomic studies have provided significant insights into elucidating P. aeruginosa evolution and its interactions with the host throughout the course of infection.
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Affiliation(s)
| | | | - Siobhán McClean
- School of Biomolecular and Biomedical Sciences, University College Dublin, Belfield, Dublin 4 D04 V1W8, Ireland; (I.J.-M.); (M.S.-M.)
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17
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Shanon MR, Al-Marzoqi AH, Hussein HJ. Prototheca spp. co-infections and their virulence factors in human protothecosis in Hillah city, Iraq. GENE REPORTS 2021. [DOI: 10.1016/j.genrep.2020.101009] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/22/2022]
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18
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Migiyama Y, Sakata S, Iyama S, Tokunaga K, Saruwatari K, Tomita Y, Saeki S, Okamoto S, Ichiyasu H, Sakagami T. Airway Pseudomonas aeruginosa density in mechanically ventilated patients: clinical impact and relation to therapeutic efficacy of antibiotics. CRITICAL CARE : THE OFFICIAL JOURNAL OF THE CRITICAL CARE FORUM 2021; 25:59. [PMID: 33573691 PMCID: PMC7876981 DOI: 10.1186/s13054-021-03488-7] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 12/11/2020] [Accepted: 02/04/2021] [Indexed: 01/31/2023]
Abstract
Background The bacterial density of Pseudomonas aeruginosa is closely related to its pathogenicity. We evaluated the effect of airway P. aeruginosa density on the clinical course of mechanically ventilated patients and the therapeutic efficacy of antibiotics. Methods We retrospectively analyzed data of mechanically ventilated ICU patients with P. aeruginosa isolated from endotracheal aspirates. Patients were divided into three groups according to the peak P. aeruginosa density during ICU stay: low (≤ 104 cfu/mL), moderate (105‒106 cfu/mL), and high (≥ 107 cfu/mL) peak density groups. The relationship between peak P. aeruginosa density and weaning from mechanical ventilation, risk factors for isolation of high peak density of P. aeruginosa, and antibiotic efficacy were investigated using multivariate and propensity score-matched analyses. Results Four-hundred-and-sixty-one patients were enrolled. Patients with high peak density of P. aeruginosa had higher inflammation and developed more severe respiratory infections. High peak density of P. aeruginosa was independently associated with few ventilator-free days on day 28 (P < 0.01) and increased ICU mortality (P = 0.047). Risk factors for high peak density of P. aeruginosa were prolonged mechanical ventilation (odd ratio [OR] 3.07 95% confidence interval [CI] 1.35‒6.97), non-antipseudomonal cephalosporins (OR 2.17, 95% CI 1.35‒3.49), hyperglycemia (OR 2.01, 95% CI 1.26‒3.22) during ICU stay, and respiratory diseases (OR 1.9, 95% CI 1.12‒3.23). Isolation of commensal colonizer was associated with lower risks of high peak density of P. aeruginosa (OR 0.43, 95% CI 0.26‒0.73). Propensity score-matched analysis revealed that antibiotic therapy for patients with ventilator-associated tracheobronchitis improved weaning from mechanical ventilation only in the high peak P. aeruginosa group. Conclusions Patients with high peak density of P. aeruginosa had worse ventilator outcome and ICU mortality. In patients with ventilator-associated tracheobronchitis, antibiotic therapy was associated with favorable ventilator weaning only in the high peak P. aeruginosa density group, and bacterial density could be a good therapeutic indicator for ventilator-associated tracheobronchitis due to P. aeruginosa.![]()
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Affiliation(s)
- Yohei Migiyama
- Department of Respiratory Medicine, Kumamoto University Hospital, Kumamoto, Japan. .,Demachi Naika, Kumamoto, Japan.
| | - Shinya Sakata
- Department of Respiratory Medicine, Kumamoto University Hospital, Kumamoto, Japan
| | - Shinji Iyama
- Department of Respiratory Medicine, Kumamoto University Hospital, Kumamoto, Japan
| | - Kentaro Tokunaga
- Department of Respiratory Medicine, Kumamoto University Hospital, Kumamoto, Japan.,Department of Critical Care Medicine, Kumamoto University Hospital, Kumamoto, Japan
| | - Koichi Saruwatari
- Department of Respiratory Medicine, Kumamoto University Hospital, Kumamoto, Japan
| | - Yusuke Tomita
- Department of Respiratory Medicine, Kumamoto University Hospital, Kumamoto, Japan
| | - Sho Saeki
- Department of Respiratory Medicine, Kumamoto University Hospital, Kumamoto, Japan
| | - Shinichiro Okamoto
- Department of Respiratory Medicine, Kumamoto University Hospital, Kumamoto, Japan
| | - Hidenori Ichiyasu
- Department of Respiratory Medicine, Kumamoto University Hospital, Kumamoto, Japan
| | - Takuro Sakagami
- Department of Respiratory Medicine, Kumamoto University Hospital, Kumamoto, Japan
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19
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Alonso B, Fernández-Barat L, Di Domenico EG, Marín M, Cercenado E, Merino I, de Pablos M, Muñoz P, Guembe M. Characterization of the virulence of Pseudomonas aeruginosa strains causing ventilator-associated pneumonia. BMC Infect Dis 2020; 20:909. [PMID: 33261585 PMCID: PMC7706020 DOI: 10.1186/s12879-020-05534-1] [Citation(s) in RCA: 48] [Impact Index Per Article: 9.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/16/2020] [Accepted: 10/21/2020] [Indexed: 12/14/2022] Open
Abstract
Background The objective of this study was to evaluate the virulence of P. aeruginosa ventilator-associated pneumonia (VAP) strains (cases) in terms of biofilm production and other phenotypic and genotypic virulence factors compared to P. aeruginosa strains isolated from other infections (controls). Methods Biofilm production was tested to assess biomass production and metabolic activity using crystal violet binding assay and XTT assay, respectively. Pigment production (pyocyanin and pyoverdine) was evaluated using cetrimide agar. Virulence genes were detected by conventional multiplex PCR and virulence was tested in an in vivo model in Galleria mellonella larvae. Results We did not find statistically significant differences between VAP and no-VAP strains (p > 0.05) regarding biofilm production. VAP strains had no production of pyocyanin after 24 h of incubation (p = 0.023). The distribution of virulence genes between both groups were similar (p > 0.05). VAP strains were less virulent than non-VAP strains in an in vivo model of G. mellonella (p < 0.001). Conclusion The virulence of VAP-Pseudomonas aeruginosa does not depend on biofilm formation, production of pyoverdine or the presence of some virulence genes compared to P. aeruginosa isolated from non-invasive locations. However, VAP strains showed attenuated virulence compared to non-VAP strains in an in vivo model of G. mellonella. Supplementary Information Supplementary information accompanies this paper at 10.1186/s12879-020-05534-1.
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Affiliation(s)
- Beatriz Alonso
- Department of Clinical Microbiology and Infectious Diseases, Hospital General Universitario Gregorio Marañón, Madrid, Spain. .,Instituto de Investigación Sanitaria Gregorio Marañón, Madrid, Spain.
| | - Laia Fernández-Barat
- Centro de Investigación Biomedica En Red-Enfermedades Respiratorias (CibeRes, CB06/06/0028) and Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain.,Center for Biomedical Research CELLEX, School of Medicine, University of Barcelona, Barcelona, Spain
| | | | - Mercedes Marín
- Department of Clinical Microbiology and Infectious Diseases, Hospital General Universitario Gregorio Marañón, Madrid, Spain
| | - Emilia Cercenado
- Department of Clinical Microbiology and Infectious Diseases, Hospital General Universitario Gregorio Marañón, Madrid, Spain
| | - Irene Merino
- Servicio de Microbiología, Hospital Universitario Ramón y Cajal and Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS), Madrid, Spain.,Red Española de Investigación en Patología Infecciosa (REIPI), Madrid, Spain.,Group For Biomedical Research in Sepsis (BioSepsis) Hospital Clínico Universitario de Valladolid, Valladolid, Spain.,Centro de Investigación Biomedica En Red - Enfermedades Respiratorias (CibeRes, CB06/06/0028), Barcelona, Spain.,National Health System, SACYL/IECSCYL, Valladolid, Spain
| | - Manuela de Pablos
- Servicio de Microbiología y Parasitología Hospital Universitario La Paz, Madrid, Spain
| | - Patricia Muñoz
- Department of Clinical Microbiology and Infectious Diseases, Hospital General Universitario Gregorio Marañón, Madrid, Spain.,Instituto de Investigación Sanitaria Gregorio Marañón, Madrid, Spain.,CIBER Enfermedades Respiratorias-CIBERES (CB06/06/0058), Madrid, Spain.,Medicine Department, School of Medicine, Universidad Complutense de Madrid, Madrid, Spain
| | - María Guembe
- Department of Clinical Microbiology and Infectious Diseases, Hospital General Universitario Gregorio Marañón, Madrid, Spain.,Instituto de Investigación Sanitaria Gregorio Marañón, Madrid, Spain
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20
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Guzzo F, Scognamiglio M, Fiorentino A, Buommino E, D’Abrosca B. Plant Derived Natural Products against Pseudomonas aeruginosa and Staphylococcus aureus: Antibiofilm Activity and Molecular Mechanisms. Molecules 2020; 25:E5024. [PMID: 33138250 PMCID: PMC7663672 DOI: 10.3390/molecules25215024] [Citation(s) in RCA: 49] [Impact Index Per Article: 9.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/30/2020] [Revised: 10/24/2020] [Accepted: 10/27/2020] [Indexed: 12/13/2022] Open
Abstract
Bacteria are social organisms able to build complex structures, such as biofilms, that are highly organized surface-associated communities of microorganisms, encased within a self- produced extracellular matrix. Biofilm is commonly associated with many health problems since its formation increases resistance to antibiotics and antimicrobial agents, as in the case of Pseudomonas aeruginosa and Staphylococcus aureus, two human pathogens causing major concern. P. aeruginosa is responsible for severe nosocomial infections, the most frequent of which is ventilator-associated pneumonia, while S. aureus causes several problems, like skin infections, septic arthritis, and endocarditis, to name just a few. Literature data suggest that natural products from plants, bacteria, fungi, and marine organisms have proven to be effective as anti-biofilm agents, inhibiting the formation of the polymer matrix, suppressing cell adhesion and attachment, and decreasing the virulence factors' production, thereby blocking the quorum sensing network. Here, we focus on plant derived chemicals, and provide an updated literature review on the anti-biofilm properties of terpenes, flavonoids, alkaloids, and phenolic compounds. Moreover, whenever information is available, we also report the mechanisms of action.
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Affiliation(s)
- Francesca Guzzo
- Dipartimento di Scienze e Tecnologie Ambientali Biologiche e Farmaceutiche–DiSTABiF, Università degli Studi della Campania “Luigi Vanvitelli”, via Vivaldi 43, I-81100 Caserta, Italy; (F.G.); (M.S.); (A.F.)
| | - Monica Scognamiglio
- Dipartimento di Scienze e Tecnologie Ambientali Biologiche e Farmaceutiche–DiSTABiF, Università degli Studi della Campania “Luigi Vanvitelli”, via Vivaldi 43, I-81100 Caserta, Italy; (F.G.); (M.S.); (A.F.)
| | - Antonio Fiorentino
- Dipartimento di Scienze e Tecnologie Ambientali Biologiche e Farmaceutiche–DiSTABiF, Università degli Studi della Campania “Luigi Vanvitelli”, via Vivaldi 43, I-81100 Caserta, Italy; (F.G.); (M.S.); (A.F.)
- Dipartimento di Biotecnologia Marina, Stazione Zoologica Anton Dohrn, Villa Comunale, 80121 Naples, Italy
| | - Elisabetta Buommino
- Dipartimento di Farmacia, Università degli Studi di Napoli “Federico II”, Via Domenico Montesano 49, 80131 Napoli, Italy;
| | - Brigida D’Abrosca
- Dipartimento di Scienze e Tecnologie Ambientali Biologiche e Farmaceutiche–DiSTABiF, Università degli Studi della Campania “Luigi Vanvitelli”, via Vivaldi 43, I-81100 Caserta, Italy; (F.G.); (M.S.); (A.F.)
- Dipartimento di Biotecnologia Marina, Stazione Zoologica Anton Dohrn, Villa Comunale, 80121 Naples, Italy
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Clinical Significance of Carbapenem-Tolerant Pseudomonas aeruginosa Isolated in the Respiratory Tract. Antibiotics (Basel) 2020; 9:antibiotics9090626. [PMID: 32967210 PMCID: PMC7558279 DOI: 10.3390/antibiotics9090626] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/31/2020] [Revised: 09/18/2020] [Accepted: 09/18/2020] [Indexed: 12/22/2022] Open
Abstract
We often come across difficult to treat infections—even after administering appropriate antibiotics according to the minimal inhibitory concentration of the causative bacteria. Antibiotic tolerance has recently started to garner attention as a crucial mechanism of refractory infections. However, few studies have reported the correlation between clinical outcomes and antibiotic tolerance. This study aims to clarify the effect of antibiotic tolerance on clinical outcomes of respiratory tract infection caused by Pseudomonas aeuginosa (P. aeruginosa). We examined a total of 63 strains isolated from sputum samples of different patients and conducted a retrospective survey with the medical records of 37 patients with imipenem-sensitive P. aeruginosa infections. Among them, we selected 15 patients with respiratory infections, and they were divided into high-tolerance minimal bactericidal concentration for adherent bacteria (MBCAD)/minimal inhibitory concentration for adherent bacteria (MICAD) ≥ 32 (n = 9) group and low-tolerance MBCAD/MICAD ≤ 16 (n = 6) group for further investigations. The findings indicated that the high-tolerance group consisted of many cases requiring hospitalization. Chest computed tomography findings showed that the disease was more extensive in the high-tolerance group compared to the low-tolerance group. Regarding the bacterial phenotypic characterization, the high-tolerance group significantly upregulated the production of the virulence factors compared to the low-tolerance group. Our study provided evidence that carbapenem tolerance level is a potent prognostic marker of P. aeruginosa infections, and carbapenem tolerance could be a potential target for new antimicrobial agents to inhibit the progression of persistent P. aeruginosa infections.
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22
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Mishra R, Kushveer JS, Khan MIK, Pagal S, Meena CK, Murali A, Dhayalan A, Venkateswara Sarma V. 2,4-Di-Tert-Butylphenol Isolated From an Endophytic Fungus, Daldinia eschscholtzii, Reduces Virulence and Quorum Sensing in Pseudomonas aeruginosa. Front Microbiol 2020; 11:1668. [PMID: 32849344 PMCID: PMC7418596 DOI: 10.3389/fmicb.2020.01668] [Citation(s) in RCA: 29] [Impact Index Per Article: 5.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/11/2020] [Accepted: 06/25/2020] [Indexed: 02/01/2023] Open
Abstract
Pseudomonas aeruginosa is among the top three gram-negative bacteria according to the WHO’s critical priority list of pathogens against which newer antibiotics are urgently needed and considered a global threat due to multiple drug resistance. This situation demands unconventional antimicrobial strategies such as the inhibition of quorum sensing to alleviate the manifestation of classical resistance mechanisms. Here, we report that 2,4-di-tert-butylphenol (2,4-DBP), isolated from an endophytic fungus, Daldinia eschscholtzii, inhibits the quorum-sensing properties of P. aeruginosa. We have found that treating P. aeruginosa with 2,4-DBP substantially reduced the secretion of virulence factors as well as biofilm, and its associated factors that are controlled by quorum sensing, in a dose-dependent manner. Concomitantly, 2,4-DBP also significantly reduced the expression of quorum sensing-related genes, i.e., lasI, lasR, rhlI, and rhlR significantly. Importantly, 2,4-DBP restricted the adhesion and invasion of P. aeruginosa to the A549 lung alveolar carcinoma cells. In addition, bactericidal assay with 2,4-DBP exhibited synergism with ampicillin to kill P. aeruginosa. Furthermore, our computational studies predicted that 2,4-DBP could bind to the P. aeruginosa quorum-sensing receptors LasR and RhlR. Collectively, these data suggest that 2,4-DBP can be exploited as a standalone drug or in combination with antibiotic(s) as an anti-virulence and anti-biofilm agent to combat the multidrug resistant P. aeruginosa infection.
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Affiliation(s)
- Rashmi Mishra
- Department of Biotechnology, Pondicherry University, Puducherry, India
| | | | - Mohd Imran K Khan
- Department of Biotechnology, Pondicherry University, Puducherry, India
| | - Sudhakar Pagal
- Department of Biotechnology, Pondicherry University, Puducherry, India
| | | | - Ayaluru Murali
- Centre for Bioinformatics, Pondicherry University, Puducherry, India
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23
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Pip serves as an intermediate in RpoS-modulated phz2 expression and pyocyanin production in Pseudomonas aeruginosa. Microb Pathog 2020; 147:104409. [PMID: 32707314 DOI: 10.1016/j.micpath.2020.104409] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/25/2020] [Revised: 07/17/2020] [Accepted: 07/17/2020] [Indexed: 11/22/2022]
Abstract
Pyocyanin, a main virulence factor that is produced by Pseudomonas aeruginosa, plays an important role in pathogen-host interaction during infection. Two copies of phenazine-biosynthetic operons on genome, phz1 (phzA1B1C1D1E1F1G1) and phz2 (phzA2B2C2D2E2F2G2), contribute to phenazine biosynthesis. In our previous study, we found that RpoS positively regulates expression of the phz2 operon and pyocyanin biosynthesis in P. aeruginosa PAO1. In this work, when a TetR-family regulator gene, pip, was knocked out, we found that pyocyanin production was dramatically reduced, indicating that Pip positively regulates pyocyanin biosynthesis. With further phenazines quantification and β-galactosidase assay, we confirmed that Pip positively regulates phz2 expression, but does not regulate phz1 expression. In addition, while the rpoS gene was deleted, expression of pip was down-regulated. Expression of rpoS in the wild-type PAO1 strain, however, was similar to that in the Pip-deficient mutant PAΔpip, suggesting that expression of pip could positively be regulated by RpoS, whereas rpoS could not be regulated by Pip. Taken together, we drew a conclusion that Pip might serve as an intermediate in RpoS-modulated expression of the phz2 operon and pyocyanin biosynthesis in P. aeruginosa.
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24
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Pourmbarak Mahnaie M, Mahmoudi H. Effect of glutathione-stabilized silver nanoparticles on expression of las I and las R of the genes in Pseudomonas aeruginosa strains. Eur J Med Res 2020; 25:17. [PMID: 32434568 PMCID: PMC7238514 DOI: 10.1186/s40001-020-00415-4] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/28/2019] [Accepted: 05/09/2020] [Indexed: 11/10/2022] Open
Abstract
Background Biofilm formation is regarded as a significant factor in the establishment of infections caused by Pseudomonas aeruginosa. P. aeruginosa is one of the most important causes of nosocomial infections. Today silver nanoparticles (Ag-NPs) are used as antimicrobials due to their well-known, chemical, biological, and physical properties. Exposure to nanoparticles could inhibit colonization of new bacteria onto the biofilm. Methods In the present work, the green synthesis of Ag-NPs was performed using the alcoholic extract of Eucalyptus camaldulensis. Ag-NPs and glutathione-stabilized silver nanoparticles (GSH–Ag-NPs) were characterized using X-ray diffraction (XRD), dynamic light scattering (DLS), scanning electron microscope (SEM), and carbon, nitrogen, and hydrogen (CNH) and Fourier transform infrared spectroscopy (FTIR) techniques were applied to investigate the structure of the modified nanoparticles. Then, the antimicrobial and antibiofilm potential of the prepared Ag-NPs and GSH–Ag-NPs against P. aeruginosa strains was evaluated using microbroth dilution method and their effects on the expression of las I and las R genes. Results In this study, a total of 50 P. aeruginosa isolates were recovered from clinical samples. According to the results, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) value of Ag-NPs against P. aeruginosa was determined to be 512–256 μg/ml, respectively, while the MIC and MBC value of GS–Ag-NPs against P. aeruginosa clinical strains was determined in a range of 128–256 μg/ml and 256–512 μg/ml, respectively. The mean expression level in las R, las I genes in P. aeruginosa strains treated with ½ MIC of Ag-NPs was decreased by −5.7 and −8fold, respectively. The mean expression levels of las R, las I genes in P. aeruginosa strains treated with ½ MIC of GS–Ag-NPs were decreased by −8.7 and −10fold, respectively (P < 0.05). Conclusions The results of our study showed that Ag-NPs and GS–Ag-NPs are highly effective against P. aeruginosa strains. Moreover, this study also proves the promising potential of using nanoparticles as anti-biofilm formation and antibacterial agents.
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Affiliation(s)
| | - Hassan Mahmoudi
- Department of Microbiology, Faculty of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran.
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25
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García-Contreras R, Loarca D, Pérez-González C, Jiménez-Cortés JG, Gonzalez-Valdez A, Soberón-Chávez G. Rhamnolipids stabilize quorum sensing mediated cooperation in Pseudomonas aeruginosa. FEMS Microbiol Lett 2020; 367:5837079. [PMID: 32407463 DOI: 10.1093/femsle/fnaa080] [Citation(s) in RCA: 13] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/22/2020] [Accepted: 05/12/2020] [Indexed: 12/24/2022] Open
Abstract
Pseudomonas aeruginosa is one of the main models to study social behaviors in bacteria since it synthesizes several exoproducts, including exoproteases and siderophores and release them to the environment. Exoproteases and siderophores are public goods that can be utilized by the individuals that produce them but also by non-producers, that are considered social cheaters. Molecularly exoprotease cheaters are mutants in regulatory genes such as lasR, and are commonly isolated from chronic infections and selected in the laboratory upon serial cultivation in media with protein as a sole carbon source. Despite that the production of exoproteases is exploitable, cooperators have also ways to restrict the growth and selection of social cheaters, for instance by producing toxic metabolites like pyocyanin. In this work, using bacterial competitions, serial cultivation and growth assays, we demonstrated that rhamnolipids which production is regulated by quorum sensing, selectively affect the growth of lasR mutants and are able to restrict social cheating, hence contributing to the maintenance of cooperation in Pseudomonas aeruginosa populations.
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Affiliation(s)
- Rodolfo García-Contreras
- Departamento de Microbiología y Parasitología, Facultad de Medicina, Universidad Nacional Autónoma de México, Circuto Escolar 411A, Copilco Universidad, Coyoacán, 04360, Mexico City, Mexico
| | - Daniel Loarca
- Departamento de Microbiología y Parasitología, Facultad de Medicina, Universidad Nacional Autónoma de México, Circuto Escolar 411A, Copilco Universidad, Coyoacán, 04360, Mexico City, Mexico
| | - Caleb Pérez-González
- Departamento de Microbiología y Parasitología, Facultad de Medicina, Universidad Nacional Autónoma de México, Circuto Escolar 411A, Copilco Universidad, Coyoacán, 04360, Mexico City, Mexico
| | - J Guillermo Jiménez-Cortés
- Departamento de Microbiología y Parasitología, Facultad de Medicina, Universidad Nacional Autónoma de México, Circuto Escolar 411A, Copilco Universidad, Coyoacán, 04360, Mexico City, Mexico
| | - Abigail Gonzalez-Valdez
- Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Tercer Circuito Exterior, Ciudad Universitaria, Apdo. Postal 70228, C. P. 04510, CDMX, México
| | - Gloria Soberón-Chávez
- Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Tercer Circuito Exterior, Ciudad Universitaria, Apdo. Postal 70228, C. P. 04510, CDMX, México
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26
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Mishra R, Kushveer JS, Khan MIK, Pagal S, Meena CK, Murali A, Dhayalan A, Venkateswara Sarma V. 2,4-Di-Tert-Butylphenol Isolated From an Endophytic Fungus, Daldinia eschscholtzii, Reduces Virulence and Quorum Sensing in Pseudomonas aeruginosa. Front Microbiol 2020; 11:1668. [PMID: 32849344 DOI: 10.3389/fmicb.2020.0166-8] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/11/2020] [Accepted: 06/25/2020] [Indexed: 05/20/2023] Open
Abstract
Pseudomonas aeruginosa is among the top three gram-negative bacteria according to the WHO's critical priority list of pathogens against which newer antibiotics are urgently needed and considered a global threat due to multiple drug resistance. This situation demands unconventional antimicrobial strategies such as the inhibition of quorum sensing to alleviate the manifestation of classical resistance mechanisms. Here, we report that 2,4-di-tert-butylphenol (2,4-DBP), isolated from an endophytic fungus, Daldinia eschscholtzii, inhibits the quorum-sensing properties of P. aeruginosa. We have found that treating P. aeruginosa with 2,4-DBP substantially reduced the secretion of virulence factors as well as biofilm, and its associated factors that are controlled by quorum sensing, in a dose-dependent manner. Concomitantly, 2,4-DBP also significantly reduced the expression of quorum sensing-related genes, i.e., lasI, lasR, rhlI, and rhlR significantly. Importantly, 2,4-DBP restricted the adhesion and invasion of P. aeruginosa to the A549 lung alveolar carcinoma cells. In addition, bactericidal assay with 2,4-DBP exhibited synergism with ampicillin to kill P. aeruginosa. Furthermore, our computational studies predicted that 2,4-DBP could bind to the P. aeruginosa quorum-sensing receptors LasR and RhlR. Collectively, these data suggest that 2,4-DBP can be exploited as a standalone drug or in combination with antibiotic(s) as an anti-virulence and anti-biofilm agent to combat the multidrug resistant P. aeruginosa infection.
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Affiliation(s)
- Rashmi Mishra
- Department of Biotechnology, Pondicherry University, Puducherry, India
| | | | - Mohd Imran K Khan
- Department of Biotechnology, Pondicherry University, Puducherry, India
| | - Sudhakar Pagal
- Department of Biotechnology, Pondicherry University, Puducherry, India
| | | | - Ayaluru Murali
- Centre for Bioinformatics, Pondicherry University, Puducherry, India
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Maurice NM, Bedi B, Yuan Z, Goldberg JB, Koval M, Hart CM, Sadikot RT. Pseudomonas aeruginosa Induced Host Epithelial Cell Mitochondrial Dysfunction. Sci Rep 2019; 9:11929. [PMID: 31417101 PMCID: PMC6695387 DOI: 10.1038/s41598-019-47457-1] [Citation(s) in RCA: 28] [Impact Index Per Article: 4.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/29/2019] [Accepted: 07/07/2019] [Indexed: 02/06/2023] Open
Abstract
The pathogenicity of P. aeruginosa is dependent on quorum sensing (QS), an inter-bacterial communication system that can also modulate host biology. The innate immune function of the lung mucosal barrier is dependent on proper mitochondrial function. The purpose of this study was to define the mechanism by which bacterial factors modulate host lung epithelial cell mitochondrial function and to investigate novel therapies that ameliorate this effect. 3-oxo-C12-HSL disrupts mitochondrial morphology, attenuates mitochondrial bioenergetics, and induces mitochondrial DNA oxidative injury. Mechanistically, we show that 3-oxo-C12-HSL attenuates expression of peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α), a master regulator of mitochondrial biogenesis, antioxidant defense, and cellular respiration, and its downstream effectors in both BEAS-2B and primary lung epithelial cells. Overexpression of PGC-1α attenuates the inhibition in cellular respiration caused by 3-oxo-C12-HSL. Pharmacologic activation of PGC-1α restores barrier integrity in cells treated with 3-oxo-C12-HSL. These data demonstrate that the P. aeruginosa QS molecule, 3-oxo-C12-HSL, alters mitochondrial pathways critical for lung mucosal immunity. Genetic and pharmacologic strategies that activate the PGC-1α pathway enhance host epithelial cell mitochondrial function and improve the epithelial innate response to P. aeruginosa. Therapies that rescue PGC-1α function may provide a complementary approach in the treatment of P. aeruginosa infection.
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Affiliation(s)
- Nicholas M Maurice
- Department of Medicine, Division of Pulmonary, Allergy, Critical Care, and Sleep Medicine, Emory University School of Medicine, Atlanta, GA, 30322, USA.,Atlanta Veterans Affairs Health Care System, Decatur, GA, 30033, USA
| | - Brahmchetna Bedi
- Department of Medicine, Division of Pulmonary, Allergy, Critical Care, and Sleep Medicine, Emory University School of Medicine, Atlanta, GA, 30322, USA.,Atlanta Veterans Affairs Health Care System, Decatur, GA, 30033, USA
| | - Zhihong Yuan
- Department of Medicine, Division of Pulmonary, Allergy, Critical Care, and Sleep Medicine, Emory University School of Medicine, Atlanta, GA, 30322, USA.,Atlanta Veterans Affairs Health Care System, Decatur, GA, 30033, USA
| | - Joanna B Goldberg
- Department of Pediatrics, Division of Pulmonology, Allergy/Immunology, Cystic Fibrosis, and Sleep, Emory University, Atlanta, GA, 30322, USA.,Children's Healthcare of Atlanta, Center for CF and Airways Disease Research Atlanta, Atlanta, GA, USA
| | - Michael Koval
- Department of Medicine, Division of Pulmonary, Allergy, Critical Care, and Sleep Medicine, Emory University School of Medicine, Atlanta, GA, 30322, USA.,Department of Cell Biology, Emory University School of Medicine, Atlanta, GA, 30322, USA
| | - C Michael Hart
- Department of Medicine, Division of Pulmonary, Allergy, Critical Care, and Sleep Medicine, Emory University School of Medicine, Atlanta, GA, 30322, USA.,Atlanta Veterans Affairs Health Care System, Decatur, GA, 30033, USA
| | - Ruxana T Sadikot
- Department of Medicine, Division of Pulmonary, Allergy, Critical Care, and Sleep Medicine, Emory University School of Medicine, Atlanta, GA, 30322, USA. .,Atlanta Veterans Affairs Health Care System, Decatur, GA, 30033, USA.
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Maurice NM, Bedi B, Sadikot RT. Pseudomonas aeruginosa Biofilms: Host Response and Clinical Implications in Lung Infections. Am J Respir Cell Mol Biol 2019; 58:428-439. [PMID: 29372812 DOI: 10.1165/rcmb.2017-0321tr] [Citation(s) in RCA: 235] [Impact Index Per Article: 39.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/11/2022] Open
Abstract
Pseudomonas aeruginosa is a major health challenge that causes recalcitrant multidrug-resistant infections, especially in immunocompromised and hospitalized patients. P. aeruginosa is an important cause of nosocomial and ventilator-associated pneumonia characterized by high prevalence and fatality rates. P. aeruginosa also causes chronic lung infections in individuals with cystic fibrosis. Multidrug- and totally drug-resistant strains of P. aeruginosa are increasing threats that contribute to high mortality in these patients. The pathogenesis of many P. aeruginosa infections depends on its ability to form biofilms, structured bacterial communities that can coat mucosal surfaces or invasive devices. These biofilms make conditions more favorable for bacterial persistence, as embedded bacteria are inherently more difficult to eradicate than planktonic bacteria. The molecular mechanisms that underlie P. aeruginosa biofilm pathogenesis and the host response to P. aeruginosa biofilms remain to be fully defined. However, it is known that biofilms offer protection from the host immune response and are also extremely recalcitrant to antimicrobial therapy. Therefore, development of novel therapeutic strategies specifically aimed at biofilms is urgently needed. Here, we review the host response, key clinical implications of P. aeruginosa biofilms, and novel therapeutic approaches to treat biofilms relevant to lung infections. Greater understanding of P. aeruginosa biofilms will elucidate novel avenues to improve outcomes for P. aeruginosa pulmonary infections.
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Affiliation(s)
- Nicholas M Maurice
- 1 Atlanta Veterans Affairs Medical Center, Decatur, Georgia; and.,2 Department of Medicine Division of Pulmonary, Allergy, Critical Care, and Sleep Medicine, Emory University, Atlanta, Georgia
| | - Brahmchetna Bedi
- 1 Atlanta Veterans Affairs Medical Center, Decatur, Georgia; and
| | - Ruxana T Sadikot
- 1 Atlanta Veterans Affairs Medical Center, Decatur, Georgia; and.,2 Department of Medicine Division of Pulmonary, Allergy, Critical Care, and Sleep Medicine, Emory University, Atlanta, Georgia
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Taha MN, Saafan AE, Ahmedy A, El Gebaly E, Khairalla AS. Two novel synthetic peptides inhibit quorum sensing-dependent biofilm formation and some virulence factors in Pseudomonas aeruginosa PAO1. J Microbiol 2019; 57:618-625. [PMID: 31054133 DOI: 10.1007/s12275-019-8548-2] [Citation(s) in RCA: 13] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/01/2018] [Revised: 01/10/2019] [Accepted: 01/30/2019] [Indexed: 11/28/2022]
Abstract
Quorum sensing (QS) regulates virulence factor expression in Pseudomonas aeruginosa. Inhibiting the QS-controlled virulence factors without inhibiting the growth of P. aeruginosa is a promising approach for overcoming the widespread resistance of P. aeruginosa. This study was proposed to investigate the effects of two novel synthetic peptides on the biofilm development and virulence factor production of P. aeruginosa. The tested strain was P. aeruginosa PAO1. The results indicated that both of the synthetic peptides (LIVRHK and LIVRRK) inhibited (P < 0.05) the formation of biofilms and the production of virulence factors, including pyocyanin, protease, and rhamnolipids, without inhibiting the growth of PAO1. Additionally, we detected transcriptional changes related to QS and found a significant reduction in the levels of gene expression of lasI, lasR, rhlI, and rhlR. This study demonstrates that LIVRRK and LIVRHK are novel synthetic peptides that can act as potent inhibitors of QS-regulated virulence factors in P. aeruginosa. Moreover, these synthetic peptides have potential applications in the treatment of biofilmrelated diseases. Both peptides may be able to control chronic infections and biofilm-associated problems of P. aeruginosa.
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Affiliation(s)
- Mostafa N Taha
- Department of Microbiology and Immunology, Faculty of Pharmacy, Nahda University, Beni-Suef, Egypt.
| | - Amal E Saafan
- Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Menoufia University, Menoufia, Egypt
| | - A Ahmedy
- Department of Microbiology and Immunology, Faculty of Pharmacy, Cairo University, Cairo, Egypt
| | - Eman El Gebaly
- Department of Microbiology and Immunology, Faculty of Pharmacy, Beni-Suef University, Beni-Suef, Egypt
| | - Ahmed S Khairalla
- Department of Microbiology and Immunology, Faculty of Pharmacy, Beni-Suef University, Beni-Suef, Egypt
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30
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Uncoupled Quorum Sensing Modulates the Interplay of Virulence and Resistance in a Multidrug-Resistant Clinical Pseudomonas aeruginosa Isolate Belonging to the MLST550 Clonal Complex. Antimicrob Agents Chemother 2019; 63:AAC.01944-18. [PMID: 30670423 DOI: 10.1128/aac.01944-18] [Citation(s) in RCA: 12] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/11/2018] [Accepted: 01/13/2019] [Indexed: 01/09/2023] Open
Abstract
Pseudomonas aeruginosa is a prevalent and pernicious pathogen equipped with extraordinary capabilities both to infect the host and to develop antimicrobial resistance (AMR). Monitoring the emergence of AMR high-risk clones and understanding the interplay of their pathogenicity and antibiotic resistance is of paramount importance to avoid resistance dissemination and to control P. aeruginosa infections. In this study, we report the identification of a multidrug-resistant (MDR) P. aeruginosa strain PA154197 isolated from a blood stream infection in Hong Kong. PA154197 belongs to a distinctive MLST550 clonal complex shared by two other international P. aeruginosa isolates VW0289 and AUS544. Comparative genome and transcriptome analysis of PA154197 with the reference strain PAO1 led to the identification of a variety of genetic variations in antibiotic resistance genes and the hyperexpression of three multidrug efflux pumps MexAB-OprM, MexEF-OprN, and MexGHI-OpmD in PA154197. Unexpectedly, the strain does not display a metabolic cost and a compromised virulence compared to PAO1. Characterizing its various physiological and virulence traits demonstrated that PA154197 produces a substantially higher level of the P. aeruginosa major virulence factor pyocyanin (PYO) than PAO1, but it produces a decreased level of pyoverdine and displays decreased biofilm formation compared with PAO1. Further analysis revealed that the secondary quorum-sensing (QS) system Pqs that primarily controls the PYO production is hyperactive in PA154197 independent of the master QS systems Las and Rhl. Together, these investigations disclose a unique, uncoupled QS mediated pathoadaptation mechanism in clinical P. aeruginosa which may account for the high pathogenic potentials and antibiotic resistance in the MDR isolate PA154197.
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31
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Persyn E, Sassi M, Aubry M, Broly M, Delanou S, Asehnoune K, Caroff N, Crémet L. Rapid genetic and phenotypic changes in Pseudomonas aeruginosa clinical strains during ventilator-associated pneumonia. Sci Rep 2019; 9:4720. [PMID: 30886315 PMCID: PMC6423012 DOI: 10.1038/s41598-019-41201-5] [Citation(s) in RCA: 21] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/29/2018] [Accepted: 02/27/2019] [Indexed: 11/24/2022] Open
Abstract
Treatment with antibiotics leads to the selection of isolates with increased resistance. We investigated if evolution towards resistance was associated with virulence changes, in the context of P. aeruginosa ventilator-associated pneumonia (VAP). Four patients were selected because they had multiple VAP episodes during short periods (12 days to 5 weeks), with emergence of resistance. We performed whole-genome sequencing of 12 P. aeruginosa from bronchoalveolar lavages or blood culture (3 isolates per patient). Production of quorum sensing-dependent virulence factors, serum resistance, cytotoxicity against A549 cells, biofilm production, and twitching motility were studied. Each patient was infected with a unique strain. For all patients, resistance development was explained by genetic events in ampD, mexR or oprD. Additional variations were detected in virulence- and/or fitness-associated genes (algB, gacA, groEL, lasR, mpl, pilE, pilM, rhlR) depending on the strain. We noticed a convergence towards quorum sensing deficiency, correlated with a decrease of pyocyanin and protease production, survival in serum, twitching motility and cytotoxicity. In one patient, changes in pilM and pilE were related to enhanced twitching. We show that the emergence of resistance in P. aeruginosa is associated with virulence modification, even in acute infections. The consequences of this short-term pathoadaptation need to be explored.
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Affiliation(s)
- Elise Persyn
- EA3826 Université de Nantes, IRS2 Nantes Biotech, Nantes Cedex 1, F-44100, France.
- CHU Nantes, 9 quai Moncousu, Nantes Cedex 1, F-44093, France.
| | - Mohamed Sassi
- Inserm U835, Université de Rennes, Rennes, F-35000, France
| | - Marc Aubry
- Université de Rennes, CNRS, IGDR [(Institut de génétique et développement de Rennes)] - UMR 6290, F-35000, Rennes, France
- Université de Rennes, Plateforme GEH, CNRS, Inserm, BIOSIT - UMS 3480, US_S 018, F-35000, Rennes, France
| | - Martin Broly
- CHU Nantes, 9 quai Moncousu, Nantes Cedex 1, F-44093, France
| | - Sandie Delanou
- EA3826 Université de Nantes, IRS2 Nantes Biotech, Nantes Cedex 1, F-44100, France
| | - Karim Asehnoune
- EA3826 Université de Nantes, IRS2 Nantes Biotech, Nantes Cedex 1, F-44100, France
- CHU Nantes, 9 quai Moncousu, Nantes Cedex 1, F-44093, France
| | - Nathalie Caroff
- EA3826 Université de Nantes, IRS2 Nantes Biotech, Nantes Cedex 1, F-44100, France
| | - Lise Crémet
- EA3826 Université de Nantes, IRS2 Nantes Biotech, Nantes Cedex 1, F-44100, France
- CHU Nantes, 9 quai Moncousu, Nantes Cedex 1, F-44093, France
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32
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Hendiani S, Pornour M, Kashef N. Quorum-sensing-regulated virulence factors in Pseudomonas aeruginosa are affected by sub-lethal photodynamic inactivation. Photodiagnosis Photodyn Ther 2019; 26:8-12. [PMID: 30753921 DOI: 10.1016/j.pdpdt.2019.02.010] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/12/2018] [Revised: 01/02/2019] [Accepted: 02/08/2019] [Indexed: 11/15/2022]
Abstract
BACKGROUND Photodynamic inactivation (PDI) is recognized as a new antimicrobial approach. It is likely that in human hosts receiving this therapy, pathogens may encounter sub-lethal doses of PDI (sPDI), which may affect microbial virulence. This study was aimed to evaluate the effect of sPDI using methylene blue (MB) on the expression of genes belonging to two quorum sensing (QS) operons (rhl and las systems) and two genes necessary for pyocyanin and rhamnolipid production (phzM and rhlA) under QS control in Pseudomonas aeruginosa. METHODS Ability of pyocyanin and rhamnolipid production of P. aeruginosa ATCC 27853 and clinical isolates exposed to sPDI (MB at 0.012 mM and light dose of 23 J/cm2 was evaluated. The effect of sPDI on expression of rhlI, rhlR, lasI, lasR, phzM and rhlA were also evaluated by quantitative real time polymerase chain reaction. RESULTS sPDI led to the down-regulation of the expression of all four QS genes (lasI, lasR, rhlI and rhlR) and rhamnolipid gene (rhlA). However, up-regulation of pyocyanin gene (phzM) was observed after sPDI. These results were consistent with phenotypic changes. CONCLUSION This study suggests that oxidative stress induced by sPDI can affect QS-regulated virulence factors of P. aeruginosa such as pyocyanin and rhamnolipids in different ways.
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Affiliation(s)
- Saghar Hendiani
- Department of Microbiology, School of Biology, College of Science, University of Tehran, Tehran, Iran.
| | - Majid Pornour
- Department of Photo Healing and Regeneration, Medical Laser Research Center, Yara Institute, Academic Center for Education, Culture and Research (ACECR), Tehran, Iran.
| | - Nasim Kashef
- Department of Microbiology, School of Biology, College of Science, University of Tehran, Tehran, Iran.
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Wang Y, Gao L, Rao X, Wang J, Yu H, Jiang J, Zhou W, Wang J, Xiao Y, Li M, Zhang Y, Zhang K, Shen L, Hua Z. Characterization of lasR-deficient clinical isolates of Pseudomonas aeruginosa. Sci Rep 2018; 8:13344. [PMID: 30190495 PMCID: PMC6127196 DOI: 10.1038/s41598-018-30813-y] [Citation(s) in RCA: 36] [Impact Index Per Article: 5.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/11/2018] [Accepted: 08/06/2018] [Indexed: 01/24/2023] Open
Abstract
Pseudomonas aeruginosa is a prevalent opportunistic pathogen that causes fatal infections in immunocompromised individuals. Quorum sensing (QS) is a cell-to-cell communication process that controls virulence gene expression and biofilm formation in P. aeruginosa. Here, the QS systems and the relevant virulence traits in clinical P. aeruginosa isolates were characterized. Eleven out of the ninety-four P. aeruginosa isolates exhibited a biofilm-deficient phenotype. Two biofilm-deficient isolates, one from blood and the one from pleural effusion, appeared to carry a same mutation in lasR. These two isolates differed in the ability to produce QS-regulated virulence factors, but contained the same functionally deficient LasR with the truncated C-terminal domains and belonged to the same multilocus sequence type (ST227). Chromosomal lasR complementation in these lasR mutants verified that lasR inactivation was the sole cause of las deficiency. LasR was not absolutely required for rhl signal in these lasR mutants, suggesting the presence of lasR-independent QS systems. We provided evidence that the virulence gene expression are not regulated in the same manner in these isolates. These results support the hypothesis that conventional QS hierarchy can be smashed by naturally occurring lasR mutation in clinical P. aeruginosa isolates and that complex QS hierarchy may play a role in maintaining infection of this opportunistic pathogen.
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Affiliation(s)
- Yao Wang
- Department of Neonatology, Children's Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation base of Child development and Critical Disorders, Chongqing Key Laboratory of Child Infection and Immunity, Chongqing, 40014, China
| | - Leiqiong Gao
- Department of Neonatology, Children's Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation base of Child development and Critical Disorders, Chongqing Key Laboratory of Child Infection and Immunity, Chongqing, 40014, China.,Department of Microbiology, Immunology, and Parasitology, Louisiana State University Health Sciences Center, New Orleans, LA, 70112, USA
| | - Xiancai Rao
- Department of Microbiology, Army Medical University, Chongqing, 400038, China
| | - Jing Wang
- Department of Neonatology, Children's Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation base of Child development and Critical Disorders, Chongqing Key Laboratory of Child Infection and Immunity, Chongqing, 40014, China
| | - Hua Yu
- Department of Central Laboratory, Xinqiao Hospital, Army Medical University, Chongqing, China
| | - Junru Jiang
- Department of Neonatology, Children's Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation base of Child development and Critical Disorders, Chongqing Key Laboratory of Child Infection and Immunity, Chongqing, 40014, China
| | - Wei Zhou
- Department of Neonatology, Children's Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation base of Child development and Critical Disorders, Chongqing Key Laboratory of Child Infection and Immunity, Chongqing, 40014, China
| | - Jin Wang
- Phase I Clinical Centre, Beijing Shijitan Hospital, Capital Medical University, Beijing, 100038, China
| | - Yonghong Xiao
- Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, the First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China, 300013
| | - Mengwen Li
- Department of Neonatology, Children's Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation base of Child development and Critical Disorders, Chongqing Key Laboratory of Child Infection and Immunity, Chongqing, 40014, China
| | - Yan Zhang
- Department of Neonatology, Children's Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation base of Child development and Critical Disorders, Chongqing Key Laboratory of Child Infection and Immunity, Chongqing, 40014, China
| | - Kebin Zhang
- Department of Central Laboratory, Xinqiao Hospital, Army Medical University, Chongqing, China
| | - Li Shen
- Department of Microbiology, Immunology, and Parasitology, Louisiana State University Health Sciences Center, New Orleans, LA, 70112, USA.
| | - Ziyu Hua
- Department of Neonatology, Children's Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation base of Child development and Critical Disorders, Chongqing Key Laboratory of Child Infection and Immunity, Chongqing, 40014, China.
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Wang K, Chen YQ, Salido MM, Kohli GS, Kong JL, Liang HJ, Yao ZT, Xie YT, Wu HY, Cai SQ, Drautz-Moses DI, Darling AE, Schuster SC, Yang L, Ding Y. The rapid in vivo evolution of Pseudomonas aeruginosa in ventilator-associated pneumonia patients leads to attenuated virulence. Open Biol 2018; 7:rsob.170029. [PMID: 28878043 PMCID: PMC5627047 DOI: 10.1098/rsob.170029] [Citation(s) in RCA: 33] [Impact Index Per Article: 4.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/07/2017] [Accepted: 07/26/2017] [Indexed: 01/15/2023] Open
Abstract
Pseudomonas aeruginosa is an opportunistic pathogen that causes severe airway infections in humans. These infections are usually difficult to treat and associated with high mortality rates. While colonizing the human airways, P. aeruginosa could accumulate genetic mutations that often lead to its better adaptability to the host environment. Understanding these evolutionary traits may provide important clues for the development of effective therapies to treat P. aeruginosa infections. In this study, 25 P. aeruginosa isolates were longitudinally sampled from the airways of four ventilator-associated pneumonia (VAP) patients. Pacbio and Illumina sequencing were used to analyse the in vivo evolutionary trajectories of these isolates. Our analysis showed that positive selection dominantly shaped P. aeruginosa genomes during VAP infections and led to three convergent evolution events, including loss-of-function mutations of lasR and mpl, and a pyoverdine-deficient phenotype. Specifically, lasR encodes one of the major transcriptional regulators in quorum sensing, whereas mpl encodes an enzyme responsible for recycling cell wall peptidoglycan. We also found that P. aeruginosa isolated at late stages of VAP infections produce less elastase and are less virulent in vivo than their earlier isolated counterparts, suggesting the short-term in vivo evolution of P. aeruginosa leads to attenuated virulence.
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Affiliation(s)
- Ke Wang
- Department of Respiratory Disease, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi, People's Republic of China.,Centre for Genomic and Personalized Medicine, Guangxi Medical University, Nanning 530021, Guangxi, People's Republic of China
| | - Yi-Qiang Chen
- Department of Respiratory Disease, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi, People's Republic of China
| | - May M Salido
- Singapore Centre for Environmental Life Sciences Engineering (SCELSE), Nanyang Technological University, Singapore 637551, Singapore
| | - Gurjeet S Kohli
- Singapore Centre for Environmental Life Sciences Engineering (SCELSE), Nanyang Technological University, Singapore 637551, Singapore
| | - Jin-Liang Kong
- Department of Respiratory Disease, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi, People's Republic of China
| | - Hong-Jie Liang
- Department of Clinical Laboratory, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi, People's Republic of China
| | - Zi-Ting Yao
- Centre for Genomic and Personalized Medicine, Guangxi Medical University, Nanning 530021, Guangxi, People's Republic of China
| | - Yan-Tong Xie
- The First Clinical School of Guangxi Medical University, Nanning 530021, Guangxi, People's Republic of China
| | - Hua-Yu Wu
- Department of Cell Biology and Genetics, Guangxi Medical University, Nanning 530021, Guangxi, People's Republic of China
| | - Shuang-Qi Cai
- Department of Respiratory Disease, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi, People's Republic of China
| | - Daniela I Drautz-Moses
- Singapore Centre for Environmental Life Sciences Engineering (SCELSE), Nanyang Technological University, Singapore 637551, Singapore
| | - Aaron E Darling
- The ithree Institute, University of Technology Sydney, Sydney, New South Wales, Australia
| | - Stephan C Schuster
- Singapore Centre for Environmental Life Sciences Engineering (SCELSE), Nanyang Technological University, Singapore 637551, Singapore.,School of Biological Sciences, Nanyang Technological University, Singapore 637551, Singapore
| | - Liang Yang
- Singapore Centre for Environmental Life Sciences Engineering (SCELSE), Nanyang Technological University, Singapore 637551, Singapore.,School of Biological Sciences, Nanyang Technological University, Singapore 637551, Singapore
| | - Yichen Ding
- Singapore Centre for Environmental Life Sciences Engineering (SCELSE), Nanyang Technological University, Singapore 637551, Singapore .,School of Biological Sciences, Nanyang Technological University, Singapore 637551, Singapore.,Interdisciplinary Graduate School, SCELSE, Nanyang Technological University, Singapore 639798, Singapore
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35
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Potential applications of biosurfactant rhamnolipids in agriculture and biomedicine. Appl Microbiol Biotechnol 2017; 101:8309-8319. [PMID: 29018916 DOI: 10.1007/s00253-017-8554-4] [Citation(s) in RCA: 76] [Impact Index Per Article: 9.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/21/2017] [Revised: 09/25/2017] [Accepted: 09/26/2017] [Indexed: 10/18/2022]
Abstract
Rhamnolipids have recently emerged as promising bioactive molecules due to their novel structures, diverse and versatile biological functions, lower toxicity, higher biodegradability, as well as production from renewable resources. The advantages of rhamnolipids make them attractive targets for research in a wide variety of applications. Especially rhamnolipids are likely to possess potential applications of the future in areas such as biomedicine, therapeutics, and agriculture. The purpose of this mini review is to provide a comprehensive prospective of biosurfactant rhamnolipids as potential antimicrobials, immune modulators, and virulence factors, and anticancer agents in the field of biomedicine and agriculture that may meet the ever-increasing future pharmacological treatment and food safety needs in human health.
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Rodrigues ME, Lopes SP, Pereira CR, Azevedo NF, Lourenço A, Henriques M, Pereira MO. Polymicrobial Ventilator-Associated Pneumonia: Fighting In Vitro Candida albicans-Pseudomonas aeruginosa Biofilms with Antifungal-Antibacterial Combination Therapy. PLoS One 2017; 12:e0170433. [PMID: 28114348 PMCID: PMC5256963 DOI: 10.1371/journal.pone.0170433] [Citation(s) in RCA: 28] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/20/2016] [Accepted: 01/04/2017] [Indexed: 11/17/2022] Open
Abstract
The polymicrobial nature of ventilator-associated pneumonia (VAP) is now evident, with mixed bacterial-fungal biofilms colonizing the VAP endotracheal tube (ETT) surface. The microbial interplay within this infection may contribute for enhanced pathogenesis and exert impact towards antimicrobial therapy. Consequently, the high mortality/morbidity rates associated to VAP and the worldwide increase in antibiotic resistance has promoted the search for novel therapeutic strategies to fight VAP polymicrobial infections. Under this scope, this work aimed to assess the activity of mono- vs combinational antimicrobial therapy using one antibiotic (Polymyxin B; PolyB) and one antifungal (Amphotericin B; AmB) agent against polymicrobial biofilms of Pseudomonas aeruginosa and Candida albicans. The action of isolated antimicrobials was firstly evaluated in single- and polymicrobial cultures, with AmB being more effective against C. albicans and PolyB against P. aeruginosa. Mixed planktonic cultures required equal or higher antimicrobial concentrations. In biofilms, only PolyB at relatively high concentrations could reduce P. aeruginosa in both monospecies and polymicrobial populations, with C. albicans displaying only punctual disturbances. PolyB and AmB exhibited a synergistic effect against P. aeruginosa and C. albicans mixed planktonic cultures, but only high doses (256 mg L-1) of PolyB were able to eradicate polymicrobial biofilms, with P. aeruginosa showing loss of cultivability (but not viability) at 2 h post-treatment, whilst C. albicans only started to be inhibited after 14 h. In conclusion, combination therapy involving an antibiotic and an antifungal agent holds an attractive therapeutic option to treat severe bacterial-fungal polymicrobial infections. Nevertheless, optimization of antimicrobial doses and further clinical pharmacokinetics/pharmacodynamics and toxicodynamics studies underpinning the optimal use of these drugs are urgently required to improve therapy effectiveness and avoid reinfection.
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Affiliation(s)
- Maria E Rodrigues
- Centre of Biological Engineering, LIBRO-Laboratório de Investigação em Biofilmes Rosário Oliveira, University of Minho, Campus de Gualtar, Braga, Portugal
| | - Susana P Lopes
- Centre of Biological Engineering, LIBRO-Laboratório de Investigação em Biofilmes Rosário Oliveira, University of Minho, Campus de Gualtar, Braga, Portugal
| | - Cláudia R Pereira
- Centre of Biological Engineering, LIBRO-Laboratório de Investigação em Biofilmes Rosário Oliveira, University of Minho, Campus de Gualtar, Braga, Portugal
| | - Nuno F Azevedo
- LEPABE-Dep. of Chemical Engineering, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, Porto, Portugal
| | - Anália Lourenço
- Departamento de Informática-Universidade de Vigo, ESEI-Escuela Superior de Ingeniería Informática, Edificio politécnico, Campus Universitario As Lagoas, Ourense, Spain.,Centre of Biological Engineering, University of Minho, Campus de Gualtar, Braga, Portugal
| | - Mariana Henriques
- Centre of Biological Engineering, LIBRO-Laboratório de Investigação em Biofilmes Rosário Oliveira, University of Minho, Campus de Gualtar, Braga, Portugal
| | - Maria O Pereira
- Centre of Biological Engineering, LIBRO-Laboratório de Investigação em Biofilmes Rosário Oliveira, University of Minho, Campus de Gualtar, Braga, Portugal
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Zheng Y, Tsuji G, Opoku-Temeng C, Sintim HO. Inhibition of P. aeruginosa c-di-GMP phosphodiesterase RocR and swarming motility by a benzoisothiazolinone derivative. Chem Sci 2016; 7:6238-6244. [PMID: 30034764 PMCID: PMC6024209 DOI: 10.1039/c6sc02103d] [Citation(s) in RCA: 33] [Impact Index Per Article: 3.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/13/2016] [Accepted: 06/15/2016] [Indexed: 01/18/2023] Open
Abstract
Various important cellular processes in bacteria are controlled by c-di-GMP, such as motility, biofilm formation and virulence factors production. C-di-GMP is synthesized from two molecules of GTP by diguanylate cyclases (DGCs) and its actions are terminated by EAL or HD-GYP domain phosphodiesterases (PDEs), which hydrolyze c-di-GMP to linear pGpG or GMP. Thus far the majority of efforts have been dedicated to the development of inhibitors of DGCs but not PDEs. This is probably because the old view was that inhibiting any c-di-GMP PDE would lead to biofilm formation, an undesirable phenotype. Recent data however suggest that some PDEs only change the localized (not global) concentration of c-di-GMP to increase bacterial virulence and do not affect biofilm formation. A challenge therefore is to be able to develop selective PDE inhibitors that inhibit virulence-associated PDEs but not inhibit PDEs that regulate bacterial biofilm formation. Using high throughput docking experiments to screen a library of 250 000 commercially available compounds against E. coli YahA (also called PdeL), a benzoisothiazolinone derivative was found to bind to the c-di-GMP binding site of YahA with favorable energetics. Paradoxically the in silico identified inhibitor (a benzoisothiazolinone derivative) did not inhibit the hydrolysis of c-di-GMP by YahA, the model PDE that was used in the docking, but instead inhibited RocR, which is a PDE from the opportunistic pathogen P. aeruginosa (PA). RocR promotes bacterial virulence but not biofilm dispersal, making it an ideal PDE to target for anti-virulence purposes. This newly identified RocR ligand displayed some selectivity and did not inhibit other P. aeruginosa PDEs, such as DipA, PvrR and PA4108. DipA, PvrR and PA4108 are key enzymes that reduce global c-di-GMP concentration and promote biofilm dispersal; therefore the identification of an inhibitor of a PA PDE, such as RocR, that does not inhibit major PDEs that modulate global c-di-GMP is an important step towards the development of selective c-di-GMP PDEs that could have interesting biomedical applications. The identified RocR ligand could also inhibit P. aeruginosa (PAO1) swarming but not swimming or biofilm formation. Rhamnolipid production was decreased, explaining the inhibition of swarming.
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Affiliation(s)
- Yue Zheng
- Department of Chemistry , Purdue University , 560 Oval Drive , West Lafayette , IN 47907 , USA .
- Center for Drug Discovery , Purdue University , 720 Clinic Drive , West Lafayette , IN 47907 , USA
- Graduate Program in Biochemistry , University of Maryland , College Park , MD 20742 , USA
| | - Genichiro Tsuji
- Department of Chemistry , Purdue University , 560 Oval Drive , West Lafayette , IN 47907 , USA .
- Center for Drug Discovery , Purdue University , 720 Clinic Drive , West Lafayette , IN 47907 , USA
| | - Clement Opoku-Temeng
- Department of Chemistry , Purdue University , 560 Oval Drive , West Lafayette , IN 47907 , USA .
- Center for Drug Discovery , Purdue University , 720 Clinic Drive , West Lafayette , IN 47907 , USA
- Graduate Program in Biochemistry , University of Maryland , College Park , MD 20742 , USA
| | - Herman O Sintim
- Department of Chemistry , Purdue University , 560 Oval Drive , West Lafayette , IN 47907 , USA .
- Center for Drug Discovery , Purdue University , 720 Clinic Drive , West Lafayette , IN 47907 , USA
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Fourie R, Ells R, Swart CW, Sebolai OM, Albertyn J, Pohl CH. Candida albicans and Pseudomonas aeruginosa Interaction, with Focus on the Role of Eicosanoids. Front Physiol 2016; 7:64. [PMID: 26955357 PMCID: PMC4767902 DOI: 10.3389/fphys.2016.00064] [Citation(s) in RCA: 57] [Impact Index Per Article: 6.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/13/2015] [Accepted: 02/11/2016] [Indexed: 12/20/2022] Open
Abstract
Candida albicans is commonly found in mixed infections with Pseudomonas aeruginosa, especially in the lungs of cystic fibrosis (CF) patients. Both of these opportunistic pathogens are able to form resistant biofilms and frequently infect immunocompromised individuals. The interaction between these two pathogens, which includes physical interaction as well as secreted factors, is mainly antagonistic. In addition, research suggests considerable interaction with their host, especially with immunomodulatory lipid mediators, termed eicosanoids. Candida albicans and Pseudomonas aeruginosa are both able to utilize arachidonic acid (AA), liberated from the host cells during infection, to form eicosanoids. The production of these eicosanoids, such as Prostaglandin E2, by the host and the pathogens may affect the dynamics of polymicrobial infection and the outcome of infections. It is of considerable importance to elucidate the role of host-produced, as well as pathogen-produced eicosanoids in polymicrobial infection. This review will focus on in vitro as well as in vivo interaction between C. albicans and P. aeruginosa, paying special attention to the role of eicosanoids in the cross-talk between host and the pathogens.
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Affiliation(s)
- Ruan Fourie
- Pathogenic Yeast Research Group, Department of Microbial, Biochemical and Food Biotechnology, University of the Free State Bloemfontein, South Africa
| | - Ruan Ells
- Pathogenic Yeast Research Group, Department of Microbial, Biochemical and Food Biotechnology, University of the Free StateBloemfontein, South Africa; National Control Laboratory, University of the Free StateBloemfontein, South Africa
| | - Chantel W Swart
- Pathogenic Yeast Research Group, Department of Microbial, Biochemical and Food Biotechnology, University of the Free State Bloemfontein, South Africa
| | - Olihile M Sebolai
- Pathogenic Yeast Research Group, Department of Microbial, Biochemical and Food Biotechnology, University of the Free State Bloemfontein, South Africa
| | - Jacobus Albertyn
- Pathogenic Yeast Research Group, Department of Microbial, Biochemical and Food Biotechnology, University of the Free State Bloemfontein, South Africa
| | - Carolina H Pohl
- Pathogenic Yeast Research Group, Department of Microbial, Biochemical and Food Biotechnology, University of the Free State Bloemfontein, South Africa
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Losa D, Köhler T, Bacchetta M, Saab JB, Frieden M, van Delden C, Chanson M. Airway Epithelial Cell Integrity Protects from Cytotoxicity of Pseudomonas aeruginosa Quorum-Sensing Signals. Am J Respir Cell Mol Biol 2015; 53:265-75. [PMID: 25562674 DOI: 10.1165/rcmb.2014-0405oc] [Citation(s) in RCA: 26] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/22/2022] Open
Abstract
Cell-to-cell communication via gap junctions regulates airway epithelial cell homeostasis and maintains the epithelium host defense. Quorum-sensing molecules produced by Pseudomonas aeruginosa coordinate the expression of virulence factors by this respiratory pathogen. These bacterial signals may also incidentally modulate mammalian airway epithelial cell responses to the pathogen, a process called interkingdom signaling. We investigated the interactions between the P. aeruginosa N-3-oxo-dodecanoyl-L-homoserine lactone (C12) quorum-sensing molecule and human airway epithelial cell gap junctional intercellular communication (GJIC). C12 degradation and its effects on cells were monitored in various airway epithelial cell models grown under nonpolarized and polarized conditions. Its concentration was further monitored in daily tracheal aspirates of colonized intubated patients. C12 rapidly altered epithelial integrity and decreased GJIC in nonpolarized airway epithelial cells, whereas other quorum-sensing molecules had no effect. The effects of C12 were dependent on [Ca(2+)]i and could be prevented by inhibitors of Src tyrosine family and Rho-associated protein kinases. In contrast, polarized airway cells grown on Transwell filters were protected from C12 except when undergoing repair after wounding. In vivo during colonization of intubated patients, C12 did not accumulate, but it paralleled bacterial densities. In vitro C12 degradation, a reaction catalyzed by intracellular paraoxonase 2 (PON2), was impaired in nonpolarized cells, whereas PON2 expression was increased during epithelial polarization. The cytotoxicity of C12 on nonpolarized epithelial cells, combined with its impaired degradation allowing its accumulation, provides an additional pathogenic mechanism for P. aeruginosa infections.
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Affiliation(s)
| | - Thilo Köhler
- 2 Service of Infectious Diseases and Department of Microbiology and Molecular Genetics, and
| | - Marc Bacchetta
- 1 Laboratory of Clinical Investigation III.,3 Department of Cell Physiology and Metabolism, Geneva University Hospitals and Medical School of the University of Geneva, Geneva, Switzerland
| | - Joanna Bou Saab
- 1 Laboratory of Clinical Investigation III.,3 Department of Cell Physiology and Metabolism, Geneva University Hospitals and Medical School of the University of Geneva, Geneva, Switzerland
| | - Maud Frieden
- 3 Department of Cell Physiology and Metabolism, Geneva University Hospitals and Medical School of the University of Geneva, Geneva, Switzerland
| | - Christian van Delden
- 2 Service of Infectious Diseases and Department of Microbiology and Molecular Genetics, and
| | - Marc Chanson
- 1 Laboratory of Clinical Investigation III.,3 Department of Cell Physiology and Metabolism, Geneva University Hospitals and Medical School of the University of Geneva, Geneva, Switzerland
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Castillo-Juárez I, Maeda T, Mandujano-Tinoco EA, Tomás M, Pérez-Eretza B, García-Contreras SJ, Wood TK, García-Contreras R. Role of quorum sensing in bacterial infections. World J Clin Cases 2015; 3:575-598. [PMID: 26244150 PMCID: PMC4517333 DOI: 10.12998/wjcc.v3.i7.575] [Citation(s) in RCA: 126] [Impact Index Per Article: 12.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 10/09/2014] [Revised: 11/30/2014] [Accepted: 04/20/2015] [Indexed: 02/05/2023] Open
Abstract
Quorum sensing (QS) is cell communication that is widely used by bacterial pathogens to coordinate the expression of several collective traits, including the production of multiple virulence factors, biofilm formation, and swarming motility once a population threshold is reached. Several lines of evidence indicate that QS enhances virulence of bacterial pathogens in animal models as well as in human infections; however, its relative importance for bacterial pathogenesis is still incomplete. In this review, we discuss the present evidence from in vitro and in vivo experiments in animal models, as well as from clinical studies, that link QS systems with human infections. We focus on two major QS bacterial models, the opportunistic Gram negative bacteria Pseudomonas aeruginosa and the Gram positive Staphylococcus aureus, which are also two of the main agents responsible of nosocomial and wound infections. In addition, QS communication systems in other bacterial, eukaryotic pathogens, and even immune and cancer cells are also reviewed, and finally, the new approaches proposed to combat bacterial infections by the attenuation of their QS communication systems and virulence are also discussed.
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Pseudomonas aeruginosa Lon and ClpXP proteases: roles in linking carbon catabolite repression system with quorum-sensing system. Curr Genet 2015; 62:1-6. [PMID: 26045103 DOI: 10.1007/s00294-015-0499-5] [Citation(s) in RCA: 24] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/19/2015] [Revised: 05/25/2015] [Accepted: 05/26/2015] [Indexed: 12/17/2022]
Abstract
Quorum sensing (QS) plays critical roles in virulence gene expression and the pathogenesis of Pseudomonas aeruginosa, an important human pathogen. However, the regulatory effects, especially that occur directly upstream of the QS system, remain largely unknown. Here, we review recent advances in the understanding of the key component of carbon catabolite repression (CCR) system and protein quality control (PQC) system in regulating the QS system in P. aeruginosa. We propose that PQC proteases Lon and ClpXP may have an important role in linking CCR with QS, and thus contribute to the integration of nutritional cues into the regulatory network governing the virulence factors expression in P. aeruginosa.
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Yang N, Ding S, Chen F, Zhang X, Xia Y, Di H, Cao Q, Deng X, Wu M, Wong CCL, Tian XX, Yang CG, Zhao J, Lan L. The Crc protein participates in down-regulation of the Lon gene to promote rhamnolipid production and rhl quorum sensing in Pseudomonas aeruginosa. Mol Microbiol 2015; 96:526-47. [PMID: 25641250 DOI: 10.1111/mmi.12954] [Citation(s) in RCA: 37] [Impact Index Per Article: 3.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 01/28/2015] [Indexed: 12/20/2022]
Abstract
Rhamnolipid acts as a virulence factor during Pseudomonas aeruginosa infection. Here, we show that deletion of the catabolite repression control (crc) gene in P. aeruginosa leads to a rhamnolipid-negative phenotype. This effect is mediated by the down-regulation of rhl quorum sensing (QS). We discover that a disruption of the gene encoding the Lon protease entirely offsets the effect of crc deletion on the production of both rhamnolipid and rhl QS signal C4-HSL. Crc is unable to bind lon mRNA in vitro in the absence of the RNA chaperon Hfq, while Crc contributes to Hfq-mediated repression of the lon gene expression at a posttranscriptional level. Deletion of crc, which results in up-regulation of lon, significantly reduces the in vivo stability and abundance of the RhlI protein that synthesizes C4-HSL, causing the attenuation of rhl QS. Lon is also capable of degrading the RhlI protein in vitro. In addition, constitutive expression of rhlI suppresses the defects of the crc deletion mutant in rhamnolipid, C4-HSL and virulence on lettuce leaves. This study therefore uncovers a novel posttranscriptional regulatory cascade, Crc-Hfq/Lon/RhlI, for the regulation of rhamnolipid production and rhl QS in P. aeruginosa.
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Affiliation(s)
- Nana Yang
- Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zuchongzhi Road, Pudong Zhangjiang Hi-Tech Park, Shanghai, 201203, China
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Olejnickova K, Hola V, Ruzicka F. Catheter-related infections caused by Pseudomonas aeruginosa: virulence factors involved and their relationships. Pathog Dis 2014; 72:87-94. [PMID: 24842562 DOI: 10.1111/2049-632x.12188] [Citation(s) in RCA: 19] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/30/2013] [Revised: 04/17/2014] [Accepted: 05/06/2014] [Indexed: 11/26/2022] Open
Abstract
The nosocomial pathogen Pseudomonas aeruginosa is equipped with a large arsenal of cell-associated and secreted virulence factors which enhance its invasive potential. The complex relationships among virulence determinants have hitherto not been fully elucidated. In the present study, 175 catheter-related isolates were observed for the presence of selected virulence factors, namely extracellular enzymes and siderophore production, biofilm formation, resistance to antibiotics, and motility. A high percentage of the strains produced most of the tested virulence factors. A positive correlation was identified between the production of several exoproducts, and also between the formation of both types of biofilm. An opposite trend was observed between the two types of biofilm and the production of siderophores. Whereas the relationship between the submerged biofilm production (i.e. the biofilm formed on the solid surface below the water level) and the siderophore secretion was negative, the production of air-liquid interface (A-L) biofilm (i.e. the biofilm floating on the surface of the cultivation medium) and the siderophore secretion were positively correlated. All correlations were statistically significant at the level P = 0.05 with the correlation coefficient γ ≥ 0.50. Our results suggest that: (1) the co-production of the lytic enzymes and siderophores can play an important role in the pathogenesis of the catheter-related infections and should be taken into account when the virulence potential is assessed; (2) biofilm-positive strains are capable of forming both submerged and non-attached A-L biofilms; and (3) the different micro-environment in the submerged biofilm and A-L biofilm layers have opposite consequences for the production of other virulence factors.
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Affiliation(s)
- Katerina Olejnickova
- Institute for Microbiology of Faculty of Medicine, Masaryk University and St. Anne's University Hospital, Brno, Czech Republic
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Laabei M, Jamieson WD, Lewis SE, Diggle SP, Jenkins ATA. A new assay for rhamnolipid detection-important virulence factors of Pseudomonas aeruginosa. Appl Microbiol Biotechnol 2014; 98:7199-209. [PMID: 24974281 DOI: 10.1007/s00253-014-5904-3] [Citation(s) in RCA: 36] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/05/2014] [Revised: 06/17/2014] [Accepted: 06/18/2014] [Indexed: 10/25/2022]
Abstract
Rhamnolipids (RLs) are heterogeneous glycolipid molecules that are composed of one or two L-rhamnose sugars and one or two β-hydroxy fatty acids, which can vary in their length and branch size. They are biosurfactants, predominantly produced by Pseudomonas aeruginosa and are important virulence factors, playing a major role in P. aeruginosa pathogenesis. Therefore, a fast, accurate and high-throughput method of detecting such molecules is of real importance. Here, we illustrate the ability to detect RL-producing P. aeruginosa strains with high sensitivity, based on an assay involving phospholipid vesicles encapsulated with a fluorescent dye. This vesicle-lysis assay is confirmed to be solely sensitive to RLs. We illustrate a half maximum concentration for vesicle lysis (EC50) of 40 μM (23.2 μg/mL) using pure commercial RLs and highlight the ability to semi-quantify RLs directly from the culture supernatant, requiring no extra extraction or processing steps or technical expertise. We show that this method is consistent with results from thin-layer chromatography detection and dry weight analysis of RLs but find that the widely used orcinol colorimetric test significantly underestimated RL quantity. Finally, we apply this methodology to compare RL production among strains isolated from either chronic or acute infections. We confirm a positive association between RL production and acute infection isolates (p = 0.0008), highlighting the role of RLs in certain infections.
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Affiliation(s)
- Maisem Laabei
- Department of Chemistry, University of Bath, Claverton Down, Bath, BA2 7AY, UK
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Alhede M, Bjarnsholt T, Givskov M, Alhede M. Pseudomonas aeruginosa biofilms: mechanisms of immune evasion. ADVANCES IN APPLIED MICROBIOLOGY 2014; 86:1-40. [PMID: 24377853 DOI: 10.1016/b978-0-12-800262-9.00001-9] [Citation(s) in RCA: 129] [Impact Index Per Article: 11.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 02/06/2023]
Abstract
The opportunistic gram-negative bacterium Pseudomonas aeruginosa is implicated in many chronic infections and is readily isolated from chronic wounds, medical devices, and the lungs of cystic fibrosis patients. P. aeruginosa is believed to persist in the host organism due to its capacity to form biofilms, which protect the aggregated, biopolymer-embedded bacteria from the detrimental actions of antibiotic treatments and host immunity. A key component in the protection against innate immunity is rhamnolipid, which is a quorum sensing (QS)-regulated virulence factor. QS is a cell-to-cell signaling mechanism used to coordinate expression of virulence and protection of aggregated biofilm cells. Rhamnolipids are known for their ability to cause hemolysis and have been shown to cause lysis of several cellular components of the human immune system, for example, macrophages and polymorphonuclear leukocytes (PMNs). In this chapter, the interplay between P. aeruginosa and the PMNs in chronic infections is discussed with focus on the role of rhamnolipids and extracellular DNA.
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Affiliation(s)
- Maria Alhede
- Department of International Health, Immunology and Microbiology, Costerton Biofilm Center, University of Copenhagen, Copenhagen, Denmark.
| | - Thomas Bjarnsholt
- Department of International Health, Immunology and Microbiology, Costerton Biofilm Center, University of Copenhagen, Copenhagen, Denmark; Department of Clinical Microbiology, Rigshospitalet, Copenhagen, Denmark
| | - Michael Givskov
- Department of International Health, Immunology and Microbiology, Costerton Biofilm Center, University of Copenhagen, Copenhagen, Denmark; Singapore Centre on Environmental Life Sciences Engineering, Nanyang Technological University, Singapore, Singapore
| | - Morten Alhede
- Department of International Health, Immunology and Microbiology, Costerton Biofilm Center, University of Copenhagen, Copenhagen, Denmark; Department of Clinical Microbiology, Rigshospitalet, Copenhagen, Denmark
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Applying insights from biofilm biology to drug development — can a new approach be developed? Nat Rev Drug Discov 2013; 12:791-808. [DOI: 10.1038/nrd4000] [Citation(s) in RCA: 348] [Impact Index Per Article: 29.0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
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Jakobsen TH, Bjarnsholt T, Jensen PØ, Givskov M, Høiby N. Targeting quorum sensing in Pseudomonas aeruginosa biofilms: current and emerging inhibitors. Future Microbiol 2013; 8:901-21. [DOI: 10.2217/fmb.13.57] [Citation(s) in RCA: 76] [Impact Index Per Article: 6.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/09/2023] Open
Abstract
Bacterial resistance to conventional antibiotics combined with an increasing acknowledgement of the role of biofilms in chronic infections has led to a growing interest in new antimicrobial strategies that target the biofilm mode of growth. In the aggregated biofilm mode, cell-to-cell communication systems involved in the process known as quorum sensing regulate coordinated expression of virulence with immune shielding mechanisms and antibiotic resistance. For two decades, the potential of interference with quorum sensing by small chemical compounds has been investigated with the aim of developing alternative antibacterial strategies. Here, we review state of the art research of quorum sensing inhibitors against the opportunistic human pathogen Pseudomonas aeruginosa, which is found in a number of biofilm-associated infections and identified as the predominant organism infecting the lungs of cystic fibrosis patients.
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Affiliation(s)
- Tim Holm Jakobsen
- Costerton Biofilm Center, Department of International Health, Immunology & Microbiology, University of Copenhagen, DK-2200 Copenhagen, Denmark
| | - Thomas Bjarnsholt
- Costerton Biofilm Center, Department of International Health, Immunology & Microbiology, University of Copenhagen, DK-2200 Copenhagen, Denmark
- Department of Clinical Microbiology, Rigshospitalet, DK-2100 Copenhagen, Denmark
| | - Peter Østrup Jensen
- Department of Clinical Microbiology, Rigshospitalet, DK-2100 Copenhagen, Denmark
| | - Michael Givskov
- Costerton Biofilm Center, Department of International Health, Immunology & Microbiology, University of Copenhagen, DK-2200 Copenhagen, Denmark
- Singapore Centre on Environmental Life Sciences Engineering, Nanyang Technological University, Singapore
| | - Niels Høiby
- Department of Clinical Microbiology, Rigshospitalet, DK-2100 Copenhagen, Denmark
- Costerton Biofilm Center, Department of International Health, Immunology & Microbiology, University of Copenhagen, DK-2200 Copenhagen, Denmark.
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Bousbia S, Raoult D, La Scola B. Pneumonia pathogen detection and microbial interactions in polymicrobial episodes. Future Microbiol 2013; 8:633-60. [DOI: 10.2217/fmb.13.26] [Citation(s) in RCA: 16] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/26/2022] Open
Abstract
Recent reports show that microbial communities associated with respiratory infections, such as pneumonia and cystic fibrosis, are more complex than expected. Most of these communities are polymicrobial and might comprise microorganisms originating from several diverse biological and ecological sources. Moreover, unexpected bacteria in the etiology of these respiratory infections have been increasingly identified. These findings were established with the use of efficient microbiological diagnostic tools, particularly molecular tools based on common gene amplification, followed by cloning and sequencing approaches, which facilitated the identification of the polymicrobial flora. Similarly, recent investigations reported that microbial interactions might exist between species in polymicrobial communities, including typical pneumonia pathogens, such as Pseudomonas aeruginosa and Candida albicans. Here, we review recent tools for microbial diagnosis, in particular, of intensive care unit pneumonia and the reported interactions between microbial species that have primarily been identified in the etiology of these infections.
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Affiliation(s)
- Sabri Bousbia
- Aix-Marseille Université, URMITE, UM 63, CNRS 7278, IRD 198, INSERM U1095, Facultés de Médecine, Marseille, France
- IHU Méditerranée Infection, Pôle des Maladies Infectieuses et Tropicales Clinique et Biologique, Fédération de Bactériologie-Hygiène-Virologie, Centre Hospitalo-Universitaire Timone, Assistance Publique – Hôpitaux de Marseille, Marseille, France
| | - Didier Raoult
- Aix-Marseille Université, URMITE, UM 63, CNRS 7278, IRD 198, INSERM U1095, Facultés de Médecine, Marseille, France
- IHU Méditerranée Infection, Pôle des Maladies Infectieuses et Tropicales Clinique et Biologique, Fédération de Bactériologie-Hygiène-Virologie, Centre Hospitalo-Universitaire Timone, Assistance Publique – Hôpitaux de Marseille, Marseille, France
| | - Bernard La Scola
- IHU Méditerranée Infection, Pôle des Maladies Infectieuses et Tropicales Clinique et Biologique, Fédération de Bactériologie-Hygiène-Virologie, Centre Hospitalo-Universitaire Timone, Assistance Publique – Hôpitaux de Marseille, Marseille, France
- Aix-Marseille Université, URMITE, UM 63, CNRS 7278, IRD 198, INSERM U1095, Facultés de Médecine, Marseille, France.
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Guan R, Ho MC, Fröhlich RFG, Tyler PC, Almo SC, Schramm VL. Methylthioadenosine deaminase in an alternative quorum sensing pathway in Pseudomonas aeruginosa. Biochemistry 2012; 51:9094-103. [PMID: 23050701 DOI: 10.1021/bi301062y] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/28/2022]
Abstract
Pseudomonas aeruginosa possesses an unusual pathway for 5'-methylthioadenosine (MTA) metabolism involving deamination to 5'-methylthioinosine (MTI) followed by N-ribosyl phosphorolysis to hypoxanthine and 5-methylthio-α-d-ribose 1-phosphate. The specific MTI phosphorylase of P. aeruginosa has been reported [Guan, R., Ho, M. C., Almo, S. C., and Schramm, V. L. (2011) Biochemistry 50, 1247-1254], and here we characterize MTA deaminase from P. aeruginosa (PaMTADA). Genomic analysis indicated the PA3170 locus to be a candidate for MTA deaminase (MTADA). Protein encoded by PA3170 was expressed and shown to deaminate MTA with 40-fold greater catalytic efficiency for MTA than for adenosine. The k(cat)/K(m) value of 1.6 × 10(7) M(-1) s(-1) for MTA is the highest catalytic efficiency known for an MTA deaminase. 5'-Methylthiocoformycin (MTCF) is a 4.8 pM transition state analogue for PaMTADA but causes no significant inhibition of human adenosine deaminase or MTA phosphorylase. MTCF is permeable to P. aeruginosa and exhibits an IC(50) of 3 nM on cellular PaMTADA activity. PaMTADA is the only activity in P. aeruginosa extracts to act on MTA. MTA and 5-methylthio-α-d-ribose are involved in quorum sensing pathways; thus, PaMTADA is a potential target for quorum sensing. The crystal structure of PaMTADA in complex with MTCF shows the transition state mimic 8(R)-hydroxyl group in contact with a catalytic site Zn(2+), the 5'-methylthio group in a hydrophobic pocket, and the transition state mimic of the diazepine ring in contact with a catalytic site Glu.
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Affiliation(s)
- Rong Guan
- Department of Biochemistry, Albert Einstein College of Medicine, Yeshiva University, 1300 Morris Park Avenue, Bronx, New York 10461, United States
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