PD biomarkers for IL-22Fc

The goal is to identify an IL-22Fc specific gene signature in human intestinal epithelial cells in order to support PD biomarkers for IL-22Fc. We have identified IL-22Fc specific activity in HT-29 cells as secreted acute phase proteins only in the presence of IL-1β. Therefore, HT-29 cells (from gCell) will be cultured with IL-22, IL-1β, IL-6 (as a pSTAT3 activation control) as well as IL-22 + IL-1β and IL-6 + IL-1β. The "SAMID" sample characteristic is a sample identifier internal to Genentech. The ID of this project in Genentech's ExpressionPlot database is PRJ0027983 Keywords: Expression profiling by array Culture HT-29 cells for 5 days in 24 wells plates. Treat cells with cytokine(s) for 24 hours at 37?C. Six groups inluce 1) Media, 2)IL-22Fc, 3)IL-6, 4)IL-1β, 5)IL-22Fc+IL-1β, 6)IL-6+IL-1β. Combine triplicate wells per sample and extract total RNA from whole blood using RNeasy kit with on column DNase treatment. Three biological replicates will be performed, each to be run on a separate chip. 1.5ug RNA at a concentration of >100ng/ml will be submitted for each sample.Culture HT-29 cells for 5 days in 24 wells plates. Treat cells with cytokine(s) for 24 hours at 37?C. Six groups inluce 1) Media, 2)IL-22Fc, 3)IL-6, 4)IL-1β, 5)IL-22Fc+IL-1β, 6)IL-6+IL-1β. Combine triplicate wells per sample and extract total RNA from whole blood using RNeasy kit with on column DNase treatment. Three biological replicates will be performed, each to be run on a separate chip. 1.5ug RNA at a concentration of >100ng/ml will be submitted for each sample.