Understanding the link between inflammasome and apoptosis through the response of THP-1 cells against drugs using droplet-based microfluidics

Abstract

Droplet-based microfluidic devices are used to investigate monocytic THP-1 cells in response to drug administration.Consistent and reproducible droplets are created, each of which acts as a bioreactor to carry out single cell experiments withminimized contamination and live cell tracking under an invertedfluorescence microscope for more than 2 days. Here, the effects ofthree different drugs (temsirolimus, rifabutin, and BAY 11-7082) on THP-1 are examined and the results are analyzed in the contextof the inflammasome and apoptosis relationship. The ASC adaptor gene tagged with GFP is monitored as the inflammasomereporter. Thus, a systematic way is presented for deciphering cell-to-cell heterogeneity, which is an important issue in cancertreatment. The drug temsirolimus, which has effects of disrupting the mTOR pathway and triggering apoptosis in tumor cells, causesTHP-1 cells to express ASC and to be involved in apoptosis. Treatment with rifabutin, which inhibits proliferation and initiatesapoptosis in cells, affects ASC expression byfirst increasing and then decreasing it. CASP-3, which has a role in apoptosis and isdirectly related to ASC, has an increasing level in inflammasome conditioning. Thus, the cell under the effect of rifabutin might befaced with programmed cell death faster. The drug BAY 11-7082, which is responsible for NF Kappa B inhibition, shows similar results totemsirolimus with more than 60% of cells having highfluorescence intensity (ASC expression). The microfluidic platform presentedhere offers strong potential for studying newly developed small-molecule inhibitors for personalized/precision medicine.