[Immuno-Oncology UK 2021] Immuno-Oncology Therapeutics Development Using PathHunter Signaling Pathway Reporter Assays
- File Name/Number:
- Immuno-Oncology UK 2021
- Year:
- 2021
Reporter genes are a well-established method used to develop cell-based assays for testing drugs that inhibit or activate targets involved in specific signaling pathways. Here, we introduce PathHunter® Signaling Pathway Reporter Assays that utilize the industry-validated Enzyme Fragment Complementation (EFC) technology to detect reporter gene activity for signaling pathways used by immuno-oncology targets.
In this poster, we show NF-κB and NFAT signaling pathway reporter assays for endogenous or heterologously-expressed target receptors. The NFAT assay is used to measure T-cell activation and has been modified to build assays interrogating PD-1 and TREM1 pathways by the addition of the relevant receptors to the complementary NFAT signaling assay cell line. We also compare the established PD-1 signaling pathway assay with its complementary PD-1 pathway reporter assay. The PD-1 signaling assay captures a proximal signaling event (via SHP-recruitment), while the PD-1 pathway reporter assay quantifies downstream effects of PD-1 (that require T-cell activation). Both assays produce robust and sensitive responses when tested with PD-1 antagonist antibodies.