STAT6 Macrophage Polarization in Response to External Fibrotic and Microplastic Stimuli Through Bioluminescence Temporal Signaturing

Abstract

Macrophages are highly plastic immune cells that regulate fibrosis, an aberrant inflammatory response resulting in the excessive buildup of connective tissue. Occurring during organ failure and chronic illness, there is a pressing need to improve the monitoring and prediction tools of immune processes that drive fibrosis. Macrophage based immune cell biosensors releasing a quantifiable bioluminescent output in response to the activation of STAT6, a reporter element linked to anti-inflammatory macrophage phenotype, would allow for temporal and spatially resolved signals to track macrophage states during the inflammatory process. My goal was to optimize a high-throughput, repeatable platform for quantifying STAT6 activation through bioluminescent temporal spectroscopy (BTS). Macrophage STAT6 activation was monitored in incubation with fibroblasts, the connective tissue cells that build up in fibrosis, in two 3D cell-culture platforms, and with nanoplastics present at high rates in the body. Application of BTS to immune response monitoring will better inform models of fibrosis and inflammation for higher in vivo translation and therapeutic intervention.