NIH Research Festival
The immune system is a network of spatially disconnected cells. Nevertheless, it exquisitely coordinates physiological responses to eliminate diverse pathogens. For such a physically disengaged system to coherently function, immune cells must possess robust systems to make context-appropriate decisions upon detecting different pathogens and pathogen loads. Our goal is to better understand how immune cells achieve such sophisticated decision making processes. NF-kappaB is the fundamental intracellular signaling pathway that is activated to initiate and modulate nearly all immune responses. We hypothesize that immune cells vary the spatiotemporal dynamic profiles of NF-kappaB family members to discriminate pathogen identity, pathogen abundance, and ultimately muster context-appropriate immune responses. Testing this hypothesis is contingent upon mechanistically and quantitatively characterizing how NF-kappaB is utilized by primary immune cells to process pathogen-specific signal information, at the single-cell level. Therefore, we are generating fluorescent protein-NF-kappaB family fusion knock-in mouse strains. The fluorescent protein cDNA sequences will be inserted, via a CRISPR-based strategy, into the endogenous NF-kappaB loci to enable real-time monitoring of endogenous NF-kappaB. Transfecting primary immune cells with expression constructs can result in low transfection efficiency, immune cell activation, and non-physiological expression levels of proteins being studied. Therefore, these models are required to study the complex dynamics of endogenous NF-kappaB, including single peaks, oscillations, and sustained nuclear localization. Using the novel mouse strains, we will dissect the dynamic information processing codes of NF-kappaB family members in response to various pathogens and pathogen loads, and investigate how primary macrophages employ these codes to make meaningful stimuli-specific decisions.
Scientific Focus Area: Systems Biology
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