Neonatal NET-Inhibitory Factor improves survival in the cecal ligation and puncture model of polymicrobial sepsis by inhibiting neutrophil extracellular traps

Frontiers in Immunology

Immunology

Michael S. Kay¹  Judah L. Evangelista¹  James M. Fulcher¹  W. Zac Stephens²  Joshua D. Schiffman³  Mark J. Cody⁴  Claudia V. de Araujo⁴  Christian C. Yost⁴  Qing Li⁵  Jacob L. Crandell⁶  John L. Rustad⁶  Frederik Denorme⁶  Robert A. Campbell⁷  Aaron C. Petrey⁸  Kimberly A. Queisser⁸ 

[1]Department of Biochemistry, University of Utah, Salt Lake City, UT, United States

[2]Department of Pathology, University of Utah, Salt Lake City, UT, United States

[3]Department of Pediatrics/Hematology-Oncology, University of Utah, Salt Lake City, UT, United States

[4]Peel Therapeutics, Inc., Salt Lake City, UT, United States

[5]Department of Pediatrics/Neonatology, University of Utah, Salt Lake City, UT, United States

[6]Molecular Medicine Program, University of Utah, Salt Lake City, UT, United States

[7]High Throughput Genomics and Bioinformatic Analysis Shared Resource, Huntsman Cancer Institute, University of Utah, Salt Lake City, UT, United States

[8]Molecular Medicine Program, University of Utah, Salt Lake City, UT, United States

[9]Molecular Medicine Program, University of Utah, Salt Lake City, UT, United States

[10]Department of Internal Medicine, University of Utah, Salt Lake City, UT, United States

[11]Molecular Medicine Program, University of Utah, Salt Lake City, UT, United States

[12]Department of Pathology, University of Utah, Salt Lake City, UT, United States

关键词: sepsis;    neutrophil;    neutrophil extracellular trap;    neonatal NET-Inhibitory Factor;    cecal ligation and puncture;    microbiome;    antibiotic resistance;    innate immunity;   

DOI  :  10.3389/fimmu.2022.1046574

 received in 2022-09-16, accepted in 2022-12-28,  发布年份 2023

来源: Frontiers

【 摘 要 】

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.

【 授权许可】

Unknown   
Copyright © 2023 de Araujo, Denorme, Stephens, Li, Cody, Crandell, Petrey, Queisser, Rustad, Fulcher, Evangelista, Kay, Schiffman, Campbell and Yost