Specific transcriptional inhibition of bone marrow-derived macrophage tumor necrosis factor-α gene expression and protein production using novel enantiomeric carbocyclic nucleoside analogues

M. Bradshaw, M. S. Rutherford, B. J. Hoeper, C. D. McWhinney, D. R. Borcherding, L. B. Schook, C. K. Edwards

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23 Scopus citations

Abstract

Tumor necrosis factor-α (TNF-α) is a powerful macrophage-derived proinflammatory cytokine, via both direct effects on host tissues as well as indirectly through the induction of other proinflammatory mediators, including interleukin- (IL) 1β and IL-6. Activation of murine bone marrow derived macrophages (BMDMφ) with lipopolysaccharide (LPS) causes rapid expression of TNF-α, which as an autocrine factor enhances BMDMφ function through IL-1β and IL-6 production. In this study, we have examined the specific transcriptional inhibition of BMDMφ TNF-α using novel enantiomeric carbocyclic nucleoside analogues, BMDMφ were derived in vitro from routine bone marrow progenitors using colony stimulating factor 1 and treated with combinations of LPS (1-100 nG/ml) and the enantiomeric carbocyclic nucleoside (10-100 μM) analogues MDL 201, 112 (9-[(1S,3R)-cis-cyclopentan-3- ol]adenine); MDL 201,451 (9-[1R,3S)-cis-cyclopentan-3-ol]adenine); MDL 201,449 (9-[(1R,3R) trans cyclopentan-3-ol]adenine) and MDL 201,484 (9- [(1S,3S)-trans cyclopentan-3-ol]adenine). Northern blot analysis showed that MDL 201,449 was the most effective agent in vitro at selectively inhibiting TNF-α. MDL 201,449 reduced TNF-α mRNA levels by nearly 50% for up to 4 hr after the simultaneous addition of LPS and the synthetic agent. In contrast, mRNA and secreted protein levels for IL-1β (measured by the D10.S bioassay) and mRNA for TNF-α receptor p60 and TNF-α receptor p80 were not significantly affected. Carbocyclic nucleoside analogues were effective when added to BMDMφup-to 2 hr after LPS treatment and at concentrations as low as 10 μM. Regulation of BMDMφ IL-6 by carbocyclic nucleoside analogues in response to LPS appears to be both concentration and time dependent, because IL-6 mRNA and secreted protein levels were inhibited at only high drug concentrations (100 μM) and effective only at longer exposure times (+4 hr of incubation) to LPS. These data support the concept that Mφ-derived proinflammatory cytokine gene expression is differentially, rather than coordinately, regulated by selective signal transduction and/or molecular pathways. Enantiomeric carbocyclic nucleoside analogues that specifically inhibit TNF-α may have therapeutic potential in inflammatory diseases, such as systemic inflammatory response syndrome, where TNF-α has been shown to have an important role in initiating the early stages of disease.

Original languageEnglish (US)
Pages (from-to)1506-1518
Number of pages13
JournalJournal of Pharmacology and Experimental Therapeutics
Volume273
Issue number3
StatePublished - Jan 1 1995

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