Neonatal NET-Inhibitory Factor improves survival in the cecal ligation and puncture model of polymicrobial sepsis by inhibiting neutrophil extracellular traps
IntroductionNeutrophil extracellular traps (NETs) clear pathogens but may contribute Q8 pathogenically to host inflammatory tissue damage during sepsis. Innovative therapeutic agents targeting NET formation and their potentially harmful collateral effects remain understudied.MethodsWe investigated a...
| Main Authors: | , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-01-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2022.1046574/full |
| _version_ | 1797857632754597888 |
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| author | Claudia V. de Araujo Claudia V. de Araujo Frederik Denorme W. Zac Stephens Qing Li Mark J. Cody Mark J. Cody Jacob L. Crandell Aaron C. Petrey Aaron C. Petrey Kimberly A. Queisser Kimberly A. Queisser John L. Rustad James M. Fulcher Judah L. Evangelista Michael S. Kay Joshua D. Schiffman Joshua D. Schiffman Robert A. Campbell Robert A. Campbell Christian C. Yost Christian C. Yost |
| author_facet | Claudia V. de Araujo Claudia V. de Araujo Frederik Denorme W. Zac Stephens Qing Li Mark J. Cody Mark J. Cody Jacob L. Crandell Aaron C. Petrey Aaron C. Petrey Kimberly A. Queisser Kimberly A. Queisser John L. Rustad James M. Fulcher Judah L. Evangelista Michael S. Kay Joshua D. Schiffman Joshua D. Schiffman Robert A. Campbell Robert A. Campbell Christian C. Yost Christian C. Yost |
| author_sort | Claudia V. de Araujo |
| collection | DOAJ |
| description | IntroductionNeutrophil extracellular traps (NETs) clear pathogens but may contribute Q8 pathogenically to host inflammatory tissue damage during sepsis. Innovative therapeutic agents targeting NET formation and their potentially harmful collateral effects remain understudied.MethodsWe investigated a novel therapeutic agent, neonatal NET-Inhibitory Factor (nNIF), in a mouse model of experimental sepsis – cecal ligation and puncture (CLP). We administered 2 doses of nNIF (1 mg/ kg) or its scrambled peptide control intravenously 4 and 10 hours after CLP treatment and assessed survival, peritoneal fluid and plasma NET formation using the MPO-DNA ELISA, aerobic bacterial colony forming units (CFU) using serial dilution and culture, peritoneal fluid and stool microbiomes using 16S rRNA gene sequencing, and inflammatory cytokine levels using a multiplexed cytokine array. Meropenem (25 mg/kg) treatment served as a clinically relevant treatment for infection.ResultsWe observed increased 6-day survival rates in nNIF (73%) and meropenem (80%) treated mice compared to controls (0%). nNIF decreased NET formation compared to controls, while meropenem did not impact NET formation. nNIF treatment led to increased peritoneal fluid and plasma bacterial CFUs consistent with loss of NET-mediated extracellular microbial killing, while nNIF treatment alone did not alter the peritoneal fluid and stool microbiomes compared to vehicle-treated CLP mice. nNIF treatment also decreased peritoneal TNF-a inflammatory cytokine levels compared to scrambled peptide control. Furthermore, adjunctive nNIF increased survival in a model of sub-optimal meropenem treatment (90% v 40%) in CLP-treated mice.DiscussionThus, our data demonstrate that nNIF inhibits NET formation in a translationally relevant mouse model of sepsis, improves survival when given as monotherapy or as an adjuvant with antibiotics, and may play an important protective role in sepsis. |
| first_indexed | 2024-04-09T20:59:51Z |
| format | Article |
| id | doaj.art-95613aabff2141a381bede33f379a8d5 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-04-09T20:59:51Z |
| publishDate | 2023-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-95613aabff2141a381bede33f379a8d52023-03-29T09:47:13ZengFrontiers Media S.A.Frontiers in Immunology1664-32242023-01-011310.3389/fimmu.2022.10465741046574Neonatal NET-Inhibitory Factor improves survival in the cecal ligation and puncture model of polymicrobial sepsis by inhibiting neutrophil extracellular trapsClaudia V. de Araujo0Claudia V. de Araujo1Frederik Denorme2W. Zac Stephens3Qing Li4Mark J. Cody5Mark J. Cody6Jacob L. Crandell7Aaron C. Petrey8Aaron C. Petrey9Kimberly A. Queisser10Kimberly A. Queisser11John L. Rustad12James M. Fulcher13Judah L. Evangelista14Michael S. Kay15Joshua D. Schiffman16Joshua D. Schiffman17Robert A. Campbell18Robert A. Campbell19Christian C. Yost20Christian C. Yost21Department of Pediatrics/Neonatology, University of Utah, Salt Lake City, UT, United StatesMolecular Medicine Program, University of Utah, Salt Lake City, UT, United StatesMolecular Medicine Program, University of Utah, Salt Lake City, UT, United StatesDepartment of Pathology, University of Utah, Salt Lake City, UT, United StatesHigh Throughput Genomics and Bioinformatic Analysis Shared Resource, Huntsman Cancer Institute, University of Utah, Salt Lake City, UT, United StatesDepartment of Pediatrics/Neonatology, University of Utah, Salt Lake City, UT, United StatesMolecular Medicine Program, University of Utah, Salt Lake City, UT, United StatesMolecular Medicine Program, University of Utah, Salt Lake City, UT, United StatesMolecular Medicine Program, University of Utah, Salt Lake City, UT, United StatesDepartment of Pathology, University of Utah, Salt Lake City, UT, United StatesMolecular Medicine Program, University of Utah, Salt Lake City, UT, United StatesDepartment of Pathology, University of Utah, Salt Lake City, UT, United StatesMolecular Medicine Program, University of Utah, Salt Lake City, UT, United StatesDepartment of Biochemistry, University of Utah, Salt Lake City, UT, United StatesDepartment of Biochemistry, University of Utah, Salt Lake City, UT, United StatesDepartment of Biochemistry, University of Utah, Salt Lake City, UT, United StatesDepartment of Pediatrics/Hematology-Oncology, University of Utah, Salt Lake City, UT, United StatesPeel Therapeutics, Inc., Salt Lake City, UT, United StatesMolecular Medicine Program, University of Utah, Salt Lake City, UT, United StatesDepartment of Internal Medicine, University of Utah, Salt Lake City, UT, United StatesDepartment of Pediatrics/Neonatology, University of Utah, Salt Lake City, UT, United StatesMolecular Medicine Program, University of Utah, Salt Lake City, UT, United StatesIntroductionNeutrophil extracellular traps (NETs) clear pathogens but may contribute Q8 pathogenically to host inflammatory tissue damage during sepsis. Innovative therapeutic agents targeting NET formation and their potentially harmful collateral effects remain understudied.MethodsWe investigated a novel therapeutic agent, neonatal NET-Inhibitory Factor (nNIF), in a mouse model of experimental sepsis – cecal ligation and puncture (CLP). We administered 2 doses of nNIF (1 mg/ kg) or its scrambled peptide control intravenously 4 and 10 hours after CLP treatment and assessed survival, peritoneal fluid and plasma NET formation using the MPO-DNA ELISA, aerobic bacterial colony forming units (CFU) using serial dilution and culture, peritoneal fluid and stool microbiomes using 16S rRNA gene sequencing, and inflammatory cytokine levels using a multiplexed cytokine array. Meropenem (25 mg/kg) treatment served as a clinically relevant treatment for infection.ResultsWe observed increased 6-day survival rates in nNIF (73%) and meropenem (80%) treated mice compared to controls (0%). nNIF decreased NET formation compared to controls, while meropenem did not impact NET formation. nNIF treatment led to increased peritoneal fluid and plasma bacterial CFUs consistent with loss of NET-mediated extracellular microbial killing, while nNIF treatment alone did not alter the peritoneal fluid and stool microbiomes compared to vehicle-treated CLP mice. nNIF treatment also decreased peritoneal TNF-a inflammatory cytokine levels compared to scrambled peptide control. Furthermore, adjunctive nNIF increased survival in a model of sub-optimal meropenem treatment (90% v 40%) in CLP-treated mice.DiscussionThus, our data demonstrate that nNIF inhibits NET formation in a translationally relevant mouse model of sepsis, improves survival when given as monotherapy or as an adjuvant with antibiotics, and may play an important protective role in sepsis.https://www.frontiersin.org/articles/10.3389/fimmu.2022.1046574/fullsepsisneutrophilneutrophil extracellular trapneonatal NET-Inhibitory Factorcecal ligation and puncturemicrobiome |
| spellingShingle | Claudia V. de Araujo Claudia V. de Araujo Frederik Denorme W. Zac Stephens Qing Li Mark J. Cody Mark J. Cody Jacob L. Crandell Aaron C. Petrey Aaron C. Petrey Kimberly A. Queisser Kimberly A. Queisser John L. Rustad James M. Fulcher Judah L. Evangelista Michael S. Kay Joshua D. Schiffman Joshua D. Schiffman Robert A. Campbell Robert A. Campbell Christian C. Yost Christian C. Yost Neonatal NET-Inhibitory Factor improves survival in the cecal ligation and puncture model of polymicrobial sepsis by inhibiting neutrophil extracellular traps Frontiers in Immunology sepsis neutrophil neutrophil extracellular trap neonatal NET-Inhibitory Factor cecal ligation and puncture microbiome |
| title | Neonatal NET-Inhibitory Factor improves survival in the cecal ligation and puncture model of polymicrobial sepsis by inhibiting neutrophil extracellular traps |
| title_full | Neonatal NET-Inhibitory Factor improves survival in the cecal ligation and puncture model of polymicrobial sepsis by inhibiting neutrophil extracellular traps |
| title_fullStr | Neonatal NET-Inhibitory Factor improves survival in the cecal ligation and puncture model of polymicrobial sepsis by inhibiting neutrophil extracellular traps |
| title_full_unstemmed | Neonatal NET-Inhibitory Factor improves survival in the cecal ligation and puncture model of polymicrobial sepsis by inhibiting neutrophil extracellular traps |
| title_short | Neonatal NET-Inhibitory Factor improves survival in the cecal ligation and puncture model of polymicrobial sepsis by inhibiting neutrophil extracellular traps |
| title_sort | neonatal net inhibitory factor improves survival in the cecal ligation and puncture model of polymicrobial sepsis by inhibiting neutrophil extracellular traps |
| topic | sepsis neutrophil neutrophil extracellular trap neonatal NET-Inhibitory Factor cecal ligation and puncture microbiome |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2022.1046574/full |
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