T3SS and alginate biosynthesis of Pseudomonas aeruginosa impair healing of infected rabbit wounds
Pseudomonas aeruginosa (a Gram-negative bacterium) is an opportunistic pathogen found in many infected wounds and is known to impair healing. To test the hypothesis that knocking out P. aeruginosa genes that are overexpressed during wound infection can cripple a pathogen's ability to impair hea...
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| Veröffentlicht in: | Microbial pathogenesis 2020-10, Vol.147 (C), p.104254-104254, Article 104254 |
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| creator | Karna, S. L. Rajasekhar Nguyen, Jesse Q. Evani, Shankar Jaikishan Qian, Li-Wu Chen, Ping Abercrombie, Johnathan J. Sebastian, Eliza A. Fourcaudot, Andrea B. Leung, Kai P. |
| description | Pseudomonas aeruginosa (a Gram-negative bacterium) is an opportunistic pathogen found in many infected wounds and is known to impair healing. To test the hypothesis that knocking out P. aeruginosa genes that are overexpressed during wound infection can cripple a pathogen's ability to impair healing, we assessed two pathways: the Type III secretion system (T3SS) and alginate biosynthesis. We generated single- and double-mutant strains of ExsA (T3SS activator), AlgD (GDP- mannose 6-dehydrogenase of alginate biosynthesis) and their complemented strains and evaluated their pathogenicity in a rabbit ear full-thickness excision-wound infection model. Wounds were inoculated with different strains (wild type, mutants, and complementary strains) at 106 CFU/wound on post-wounding day 3. After 24 h, 5 days and 9 days post-infection, wounds were harvested for measuring bacterial counts (viable and total) and wound healing (epithelial gap). On day 9 post-infection, the viable counts of the double mutant, (exsA/algD)‾ were 100-fold lower than the counts of the wild type (PAO1), single mutants, or the complement double-mutant, (exsA/algD)‾/+. Also, when compared to wounds infected with wild type or control strains, wounds infected with the double-knockout mutant was less inhibitory to wound healing (p |
| doi_str_mv | 10.1016/j.micpath.2020.104254 |
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•Deletion of highly expressed P.aeruginosa genes in wound infection cripples bacterium pathogenicity & improves wound healing.•P. aeruginosa strain lacking virulence factor, ExsA & biofilm polysaccharide, AlgD reduced the fitness & survival in wounds.•Anti-pathogenic approach reduces selective pressure for microbial resistance and improves wound healing.</description><identifier>ISSN: 0882-4010</identifier><identifier>EISSN: 1096-1208</identifier><identifier>DOI: 10.1016/j.micpath.2020.104254</identifier><identifier>PMID: 32416139</identifier><language>eng</language><publisher>England: Elsevier Ltd</publisher><subject>Alginate ; Biofilm ; Pseudomonas aeruginosa ; Rabbit ear wound healing model ; Type III secretion System ; Virulence</subject><ispartof>Microbial pathogenesis, 2020-10, Vol.147 (C), p.104254-104254, Article 104254</ispartof><rights>2020 The Authors</rights><rights>Copyright © 2020. Published by Elsevier Ltd.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c439t-fb24d957d66b5ea40cbb1efc13fd7117893494f20a4c376a402c3f6e73bf073c3</citedby><cites>FETCH-LOGICAL-c439t-fb24d957d66b5ea40cbb1efc13fd7117893494f20a4c376a402c3f6e73bf073c3</cites><orcidid>0000-0003-3135-3653 ; 0000-0001-5042-1434 ; 0000000331353653 ; 0000000150421434</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.micpath.2020.104254$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>230,314,777,781,882,3537,27905,27906,45976</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/32416139$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://www.osti.gov/biblio/1618359$$D View this record in Osti.gov$$Hfree_for_read</backlink></links><search><creatorcontrib>Karna, S. L. Rajasekhar</creatorcontrib><creatorcontrib>Nguyen, Jesse Q.</creatorcontrib><creatorcontrib>Evani, Shankar Jaikishan</creatorcontrib><creatorcontrib>Qian, Li-Wu</creatorcontrib><creatorcontrib>Chen, Ping</creatorcontrib><creatorcontrib>Abercrombie, Johnathan J.</creatorcontrib><creatorcontrib>Sebastian, Eliza A.</creatorcontrib><creatorcontrib>Fourcaudot, Andrea B.</creatorcontrib><creatorcontrib>Leung, Kai P.</creatorcontrib><title>T3SS and alginate biosynthesis of Pseudomonas aeruginosa impair healing of infected rabbit wounds</title><title>Microbial pathogenesis</title><addtitle>Microb Pathog</addtitle><description>Pseudomonas aeruginosa (a Gram-negative bacterium) is an opportunistic pathogen found in many infected wounds and is known to impair healing. To test the hypothesis that knocking out P. aeruginosa genes that are overexpressed during wound infection can cripple a pathogen's ability to impair healing, we assessed two pathways: the Type III secretion system (T3SS) and alginate biosynthesis. We generated single- and double-mutant strains of ExsA (T3SS activator), AlgD (GDP- mannose 6-dehydrogenase of alginate biosynthesis) and their complemented strains and evaluated their pathogenicity in a rabbit ear full-thickness excision-wound infection model. Wounds were inoculated with different strains (wild type, mutants, and complementary strains) at 106 CFU/wound on post-wounding day 3. After 24 h, 5 days and 9 days post-infection, wounds were harvested for measuring bacterial counts (viable and total) and wound healing (epithelial gap). On day 9 post-infection, the viable counts of the double mutant, (exsA/algD)‾ were 100-fold lower than the counts of the wild type (PAO1), single mutants, or the complement double-mutant, (exsA/algD)‾/+. Also, when compared to wounds infected with wild type or control strains, wounds infected with the double-knockout mutant was less inhibitory to wound healing (p < 0.05). Additionally, the double mutant showed greater susceptibility to macrophage phagocytosis in vitro than all other strains (p < 0.001). In conclusion, compared to single gene knockouts, double knockout of virulence genes in T3SS pathway and alginate biosynthesis pathway is more effective in reducing P. aeruginosa pathogenicity and its ability to impair wound healing. This study highlights the necessity of a dual-targeted anti-virulence strategy to improve healing outcomes of P. aeruginosa-infected wounds.
•Deletion of highly expressed P.aeruginosa genes in wound infection cripples bacterium pathogenicity & improves wound healing.•P. aeruginosa strain lacking virulence factor, ExsA & biofilm polysaccharide, AlgD reduced the fitness & survival in wounds.•Anti-pathogenic approach reduces selective pressure for microbial resistance and improves wound healing.</description><subject>Alginate</subject><subject>Biofilm</subject><subject>Pseudomonas aeruginosa</subject><subject>Rabbit ear wound healing model</subject><subject>Type III secretion System</subject><subject>Virulence</subject><issn>0882-4010</issn><issn>1096-1208</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><recordid>eNqFkU2LFDEQhoMo7uzqT1CCJy895qu_TiKLusKCwq7nkI_KTobuZEzSK_vvTdOjV6lDQfFU1Vv1IvSGkj0ltPtw3M_enFQ57Blha02wVjxDO0rGrqGMDM_RjgwDawSh5AJd5nwkhIyCjy_RBWeCdpSPO6Tu-d0dVsFiNT34oApg7WN-CuUA2WccHf6RYbFxjkFlrCAtFYtZYT-flE_4AGry4WEFfXBgCliclNa-4N9xCTa_Qi-cmjK8Pucr9PPL5_vrm-b2-9dv159uG1M1lcZpJuzY9rbrdAtKEKM1BWcod7antB9GLkbhGFHC8L6rADPcddBz7UjPDb9C77a5MRcvs_EFzMHEEKomWa8deDtW6P0GnVL8tUAucvbZwDSpAHHJkglSgw1tX9F2Q02KOSdw8pT8rNKTpESuFsijPFsgVwvkZkHte3tesegZ7L-uvz-vwMcNgPqNRw9pFQvBgPVp1Wqj_8-KP28wmk4</recordid><startdate>20201001</startdate><enddate>20201001</enddate><creator>Karna, S. 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Rajasekhar ; Nguyen, Jesse Q. ; Evani, Shankar Jaikishan ; Qian, Li-Wu ; Chen, Ping ; Abercrombie, Johnathan J. ; Sebastian, Eliza A. ; Fourcaudot, Andrea B. ; Leung, Kai P.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c439t-fb24d957d66b5ea40cbb1efc13fd7117893494f20a4c376a402c3f6e73bf073c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Alginate</topic><topic>Biofilm</topic><topic>Pseudomonas aeruginosa</topic><topic>Rabbit ear wound healing model</topic><topic>Type III secretion System</topic><topic>Virulence</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Karna, S. L. Rajasekhar</creatorcontrib><creatorcontrib>Nguyen, Jesse Q.</creatorcontrib><creatorcontrib>Evani, Shankar Jaikishan</creatorcontrib><creatorcontrib>Qian, Li-Wu</creatorcontrib><creatorcontrib>Chen, Ping</creatorcontrib><creatorcontrib>Abercrombie, Johnathan J.</creatorcontrib><creatorcontrib>Sebastian, Eliza A.</creatorcontrib><creatorcontrib>Fourcaudot, Andrea B.</creatorcontrib><creatorcontrib>Leung, Kai P.</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>OSTI.GOV</collection><jtitle>Microbial pathogenesis</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Karna, S. L. Rajasekhar</au><au>Nguyen, Jesse Q.</au><au>Evani, Shankar Jaikishan</au><au>Qian, Li-Wu</au><au>Chen, Ping</au><au>Abercrombie, Johnathan J.</au><au>Sebastian, Eliza A.</au><au>Fourcaudot, Andrea B.</au><au>Leung, Kai P.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>T3SS and alginate biosynthesis of Pseudomonas aeruginosa impair healing of infected rabbit wounds</atitle><jtitle>Microbial pathogenesis</jtitle><addtitle>Microb Pathog</addtitle><date>2020-10-01</date><risdate>2020</risdate><volume>147</volume><issue>C</issue><spage>104254</spage><epage>104254</epage><pages>104254-104254</pages><artnum>104254</artnum><issn>0882-4010</issn><eissn>1096-1208</eissn><abstract>Pseudomonas aeruginosa (a Gram-negative bacterium) is an opportunistic pathogen found in many infected wounds and is known to impair healing. To test the hypothesis that knocking out P. aeruginosa genes that are overexpressed during wound infection can cripple a pathogen's ability to impair healing, we assessed two pathways: the Type III secretion system (T3SS) and alginate biosynthesis. We generated single- and double-mutant strains of ExsA (T3SS activator), AlgD (GDP- mannose 6-dehydrogenase of alginate biosynthesis) and their complemented strains and evaluated their pathogenicity in a rabbit ear full-thickness excision-wound infection model. Wounds were inoculated with different strains (wild type, mutants, and complementary strains) at 106 CFU/wound on post-wounding day 3. After 24 h, 5 days and 9 days post-infection, wounds were harvested for measuring bacterial counts (viable and total) and wound healing (epithelial gap). On day 9 post-infection, the viable counts of the double mutant, (exsA/algD)‾ were 100-fold lower than the counts of the wild type (PAO1), single mutants, or the complement double-mutant, (exsA/algD)‾/+. Also, when compared to wounds infected with wild type or control strains, wounds infected with the double-knockout mutant was less inhibitory to wound healing (p < 0.05). Additionally, the double mutant showed greater susceptibility to macrophage phagocytosis in vitro than all other strains (p < 0.001). In conclusion, compared to single gene knockouts, double knockout of virulence genes in T3SS pathway and alginate biosynthesis pathway is more effective in reducing P. aeruginosa pathogenicity and its ability to impair wound healing. This study highlights the necessity of a dual-targeted anti-virulence strategy to improve healing outcomes of P. aeruginosa-infected wounds.
•Deletion of highly expressed P.aeruginosa genes in wound infection cripples bacterium pathogenicity & improves wound healing.•P. aeruginosa strain lacking virulence factor, ExsA & biofilm polysaccharide, AlgD reduced the fitness & survival in wounds.•Anti-pathogenic approach reduces selective pressure for microbial resistance and improves wound healing.</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>32416139</pmid><doi>10.1016/j.micpath.2020.104254</doi><tpages>1</tpages><orcidid>https://orcid.org/0000-0003-3135-3653</orcidid><orcidid>https://orcid.org/0000-0001-5042-1434</orcidid><orcidid>https://orcid.org/0000000331353653</orcidid><orcidid>https://orcid.org/0000000150421434</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Alginate Biofilm Pseudomonas aeruginosa Rabbit ear wound healing model Type III secretion System Virulence |
| title | T3SS and alginate biosynthesis of Pseudomonas aeruginosa impair healing of infected rabbit wounds |
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