Mechanistic analysis of macrophage response to IRAK-1 gene knockdown by a smart polymer-antisense oligonucleotide therapeutic.
An excessive inflammatory response is a clinical problem following major infections and severe injury that may lead to Sepsis Syndrome and Multiple Organ Failure (MOF), including the Acute Respiratory Distress Syndrome (ARDS). Management of excessive inflammation may be possible through control of key inflammatory pathways such as those mediated by the important interleukin-1 receptor associated kinase-1 (IRAK-1). In the current study, we report the impact on gene expression induced by lipopolysaccharide (LPS) stimulation of THP-1 cells treated with an antisense oligonucleotide (ASODN) against the IRAK-1 gene using cDNA microarrays and quantitative RT-PCR. The therapeutic ASODN was delivered using a pH-sensitive, membrane-interactive polymer that destabilizes the endosomal membrane to enhance access cytoplasmic delivery in targeted cells. Following LPS stimulation, the anti-inflammatory activity of ASODN against the IRAK-1 gene expression is evidenced by the lower expression of inflammatory chemokines, cytokines and acute-phase proteins compared to control cells. These results provide a larger mechanistic picture of IRAK-1 knockdown by this polymer therapeutic in macrophage-like cells.
Johns
Rachel E
RE
Department of Bioengineering, University of Washington, Seattle, WA 98195, USA.
El-Sayed
Mohamed E H
ME
Bulmus
Volga
V
Cuschieri
Joseph
J
Maier
Ronald
R
Hoffman
Allan S
AS
Stayton
Patrick S
PS
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D045744:Cell Line, Tumor; D018925:Chemokines / Q000235:genetics / Q000378:metabolism; D003593:Cytoplasm / Q000235:genetics / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D055785:Gene Knockdown Techniques; D006801:Humans; D007249:Inflammation / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism; D053592:Interleukin-1 Receptor-Associated Kinases / Q000235:genetics*; D007378:Interleukins / Q000235:genetics / Q000378:metabolism; D008070:Lipopolysaccharides / Q000235:genetics / Q000378:metabolism; D008264:Macrophages / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D008668:Metallothionein / Q000235:genetics / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D020319:Oligodeoxyribonucleotides, Antisense / Q000235:genetics / Q000494:pharmacology* / Q000627:therapeutic use*; D020411:Oligonucleotide Array Sequence Analysis; D011108:Polymers / Q000138:chemical synthesis / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction
D018925:Chemokines; D018076:DNA, Complementary; D007378:Interleukins; D008070:Lipopolysaccharides; D016328:NF-kappa B; D020319:Oligodeoxyribonucleotides, Antisense; D011108:Polymers; D008668:Metallothionein; D053592:Interleukin-1 Receptor-Associated Kinases