A broad kinase inhibitor library live cell imaging screen using liver, kidney and cardiac hiPSC-ICAM1-eGFP reporter lineages identifies liabilities for TNFα pathway modulation.

Kinase inhibitors (KIs) are an increasing class of drugs targeting protein kinase activity in various pathologies, but predominantly in cancer and autoimmune diseases. Despite successful clinical applications, one of the challenges that remains is their safety, since many KIs lack selectivity and inhibit multi kinases or block pathways that are also important for homeostasis of healthy cells. This may lead to adverse reactions in several critical target organs of toxicity, including heart, liver and kidney. Inflammatory cytokine-mediated intracellular signaling is critical for the control of cell survival and adverse outcome and are modulated by diverse kinases. Likewise, inhibition of kinase activities by on- or off-target effects of KIs may contribute to adverse effects. In this study we systematically screened 760 different KIs targeting the majority of the kinome for their ability to affect TNFα-mediated expression of ICAM1 in hiPSC-derived renal proximal tubule-like cells (PTLCs), cardiomyocytes-like cells (CMLC) and hepatocyte-like cells (HLC). Palbociclib (CDK4/6), miltefosine (PI3K/AKT/mTOR), gilteritinib (FLT3/AXL), and erdafitinib (FGFR) led to increased expression of the ICAM1-eGFP reporter and the release of chemokines. Kinase inhibitor activity data from the ChEMBL database indicated off-target kinase inhibitor activity associated with enhanced ICAM1-eGFP expression. Finally, clinically relevant KIs that enhanced ICAM1-eGFP expression in PTLCs were found to be associated with renal adverse events in patients. Our overall findings support the application of imaging-based hiPSC-derived ICAM1-eGFP reporter covering different differentiated cell lineages to identify liabilities of novel kinase inhibitors to modulate TNFα pathways.