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A High Content Screen in Macrophages Identifies Small Molecule Modulators of STING-IRF3 and NFkB Signaling
journal contribution
posted on 2018-03-16, 21:48 authored by Peter D. Koch, Howard R. Miller, Gary Yu, John A. Tallarico, Peter K. Sorger, Yuan Wang, Yan Feng, Jason R. Thomas, Nathan T. Ross, Timothy MitchisonWe
screened a library of bioactive small molecules for activators
and inhibitors of innate immune signaling through IRF3 and NFkB pathways
with the goals of advancing pathway understanding and discovering
probes for immunology research. We used high content screening to
measure the translocation from the cytoplasm to nucleus of IRF3 and
NFkB in primary human macrophages; these transcription factors play
a critical role in the activation of STING and other pro-inflammatory
pathways. Our pathway activator screen yielded a diverse set of hits
that promoted nuclear translocation of IRF3 and/or NFkB, but the majority
of these compounds did not cause activation of downstream pathways.
Screening for antagonists of the STING pathway yielded multiple kinase
inhibitors, some of which inhibit kinases not previously known to
regulate the activity of this pathway. Structure–activity relationships
(SARs) and subsequent chemical proteomics experiments suggested that
MAPKAPK5 (PRAK) is a kinase that regulates IRF3 translocation in human
macrophages. Our work establishes a high content screening approach
for measuring pro-inflammatory pathways in human macrophages and identifies
novel ways to inhibit such pathways; among the targets of the screen
are several molecules that may merit further development as anti-inflammatory
drugs.