Intranuclear interactomic inhibition of NF-κB suppresses LPS-induced severe sepsis

Biochem Biophys Res Commun. 2015 Aug 28;464(3):711-7. doi: 10.1016/j.bbrc.2015.07.008. Epub 2015 Jul 6.

Abstract

Suppression of nuclear factor-κB (NF-κB) activation, which is best known as a major regulator of innate and adaptive immune responses, is a potent strategy for the treatment of endotoxic sepsis. To inhibit NF-κB functions, we designed the intra-nuclear transducible form of transcription modulation domain (TMD) of RelA (p65), called nt-p65-TMD, which can be delivered effectively into the nucleus without influencing the cell viability, and work as interactomic inhibitors via disruption of the endogenous p65-mediated transcription complex. nt-p65-TMD effectively inhibited the secretion of pro-inflammatory cytokines, including TNF-α, IL-1β, or IL-6 from BV2 microglia cells stimulated by lipopolysaccharide (LPS). nt-p65-TMD did not inhibit tyrosine phosphorylation of signaling mediators such as ZAP-70, p38, JNK, or ERK involved in T cell activation, but was capable of suppressing the transcriptional activity of NF-κB without the functional effect on that of NFAT upon T-cell receptor (TCR) stimulation. The transduced nt-p65-TMD in T cell did not affect the expression of CD69, however significantly inhibited the secretion of T cell-specific cytokines such as IL-2, IFN-γ, IL-4, IL-17A, or IL-10. Systemic administration of nt-p65-TMD showed a significant therapeutic effect on LPS-induced sepsis model by inhibiting pro-inflammatory cytokines secretion. Therefore, nt-p65-TMD can be a novel therapeutics for the treatment of various inflammatory diseases, including sepsis, where a transcription factor has a key role in pathogenesis, and further allows us to discover new functions of p65 under normal physiological condition without genetic alteration.

Keywords: Inflammation; Intra-nucleus delivery; NF-κB; RelA (p65); Sepsis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cells, Cultured
  • Cytokines / antagonists & inhibitors
  • Cytokines / biosynthesis
  • Female
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Jurkat Cells
  • Lipopolysaccharides / toxicity
  • Lymphocyte Activation
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • NF-kappa B / antagonists & inhibitors*
  • NF-kappa B / genetics
  • NF-kappa B / immunology
  • Protein Structure, Tertiary / genetics
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Recombinant Proteins / pharmacology
  • Sepsis / etiology
  • Sepsis / metabolism
  • Sepsis / therapy*
  • T-Lymphocytes / immunology
  • T-Lymphocytes / metabolism
  • Transcription Factor RelA / antagonists & inhibitors
  • Transcription Factor RelA / genetics
  • Transcription Factor RelA / immunology
  • Transcription, Genetic
  • Transduction, Genetic

Substances

  • Cytokines
  • Lipopolysaccharides
  • NF-kappa B
  • RELA protein, human
  • Recombinant Proteins
  • Rela protein, mouse
  • Transcription Factor RelA