title: The "genomic storm" induced by bacterial endotoxin is calmed by a nuclear transport modifier that attenuates localized and systemic inflammation.
abstract: Lipopolysaccharide (LPS) is a potent microbial virulence factor that can trigger production of proinflammatory mediators involved in the pathogenesis of localized and systemic inflammation. Importantly, the role of nuclear transport of stress responsive transcription factors in this LPS-generated "genomic storm" remains largely undefined. We developed a new nuclear transport modifier (NTM) peptide, cell-penetrating cSN50.1, which targets nuclear transport shuttles importin α5 and importin β1, to analyze its effect in LPS-induced localized (acute lung injury) and systemic (lethal endotoxic shock) murine inflammation models. We analyzed a human genome database to match 46 genes that encode cytokines, chemokines and their receptors with transcription factors whose nuclear transport is known to be modulated by NTM. We then tested the effect of cSN50.1 peptide on proinflammatory gene expression in murine bone marrow-derived macrophages stimulated with LPS. This NTM suppressed a proinflammatory transcriptome of 37 out of 84 genes analyzed, without altering expression of housekeeping genes or being cytotoxic. Consistent with gene expression analysis in primary macrophages, plasma levels of 23 out of 26 LPS-induced proinflammatory cytokines, chemokines, and growth factors were significantly attenuated in a murine model of LPS-induced systemic inflammation (lethal endotoxic shock) while the anti-inflammatory cytokine, interleukin 10, was enhanced. This anti-inflammatory reprogramming of the endotoxin-induced genomic response was accompanied by complete protection against lethal endotoxic shock with prophylactic NTM treatment, and 75% protection when NTM was first administered after LPS exposure. In a murine model of localized lung inflammation caused by direct airway exposure to LPS, expression of cytokines and chemokines in the bronchoalveolar space was suppressed with a concomitant reduction of neutrophil trafficking. Thus, calming the LPS-triggered "genomic storm" by modulating nuclear transport with cSN50.1 peptide attenuates the systemic inflammatory response associated with lethal shock as well as localized lung inflammation.
authors.affiliation: Department of Microbiology and Immunology Vanderbilt University School of Medicine, Nashville, Tennessee, United States of America.
authors.forename: Antonio
authors.initials: A
authors.lastname: DiGiandomenico
authors.affiliation: Immunotherapy Program at Vanderbilt University School of Medicine, Nashville, Tennessee, United States of America; Department of Medicine, Division of Allergy, Pulmonary and Critical Care Medicine, Vanderbilt University School of Medicine, Nashville, Tennessee, United States of America.
authors.forename: Ruth Ann
authors.initials: RA
authors.lastname: Veach
authors.affiliation: Immunotherapy Program at Vanderbilt University School of Medicine, Nashville, Tennessee, United States of America; Department of Medicine, Division of Allergy, Pulmonary and Critical Care Medicine, Vanderbilt University School of Medicine, Nashville, Tennessee, United States of America.
authors.forename: Jozef
authors.initials: J
authors.lastname: Zienkiewicz
authors.affiliation: Immunotherapy Program at Vanderbilt University School of Medicine, Nashville, Tennessee, United States of America; Department of Pediatrics, Ian Burr Division of Endocrinology and Diabetes, Vanderbilt University School of Medicine, Nashville, Tennessee, United States of America; Department of Pathology, Microbiology and Immunology, Vanderbilt University School of Medicine, Nashville, Tennessee, United States of America.
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Moore
authors.affiliation: Immunotherapy Program at Vanderbilt University School of Medicine, Nashville, Tennessee, United States of America; Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee, United States of America.
authors.forename: Lukasz S
authors.initials: LS
authors.lastname: Wylezinski
authors.affiliation: Department of Microbiology and Immunology Vanderbilt University School of Medicine, Nashville, Tennessee, United States of America; Immunotherapy Program at Vanderbilt University School of Medicine, Nashville, Tennessee, United States of America.
authors.forename: Martha A
authors.initials: MA
authors.lastname: Hutchens
authors.affiliation: Immunotherapy Program at Vanderbilt University School of Medicine, Nashville, Tennessee, United States of America; Department of Medicine, Division of Allergy, Pulmonary and Critical Care Medicine, Vanderbilt University School of Medicine, Nashville, Tennessee, United States of America; Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee, United States of America.
authors.forename: Jacek
authors.initials: J
authors.lastname: Hawiger
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000187:drug effects; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D002467:Cell Nucleus / Q000187:drug effects* / Q000378:metabolism; D057846:Cell-Penetrating Peptides / Q000494:pharmacology* / Q000627:therapeutic use; D018925:Chemokines / Q000097:blood; D005786:Gene Expression Regulation / Q000187:drug effects; D015894:Genome, Human / Q000187:drug effects / Q000235:genetics*; D006801:Humans; D008070:Lipopolysaccharides / Q000633:toxicity*; D008264:Macrophages / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D011014:Pneumonia / Q000139:chemically induced / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D012772:Shock, Septic / Q000139:chemically induced / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D014157:Transcription Factors / Q000378:metabolism; D059467:Transcriptome / Q000187:drug effects*
chemical_list: D057846:Cell-Penetrating Peptides; D018925:Chemokines; D008070:Lipopolysaccharides; D014157:Transcription Factors