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Updated: Jan 12, 2026
A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells
Published on: July 3, 2013
A broad kinase inhibitor library live cell imaging screen using liver, kidney and cardiac hiPSC-ICAM1-eGFP reporter
Matthijs Vlasveld1, Linda van den Berk1, Janet Piñero2
1Division of Cell Systems and Drug Safety, Leiden Academic Centre for Drug Research (LACDR), Leiden University, Einsteinweg 55, 2333CC, Leiden, The Netherlands.
Abstract:
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.
Insights
This study screened 760 kinase inhibitors (KIs) and found that some KIs can increase ICAM1 expression, potentially causing organ toxicity. Clinically used KIs linked to kidney damage were identified, highlighting a new method for predicting KI safety.
Area of Science:
- Pharmacology
- Toxicology
- Cell Biology
Background:
- Kinase inhibitors (KIs) are crucial therapeutics for cancer and autoimmune diseases.
- KI selectivity remains a challenge, leading to potential toxicity in organs like the heart, liver, and kidney.
- Tumor necrosis factor-alpha (TNFα)-mediated signaling, modulated by kinases, impacts cell survival and adverse outcomes.
Purpose of the Study:
- To systematically screen a large panel of kinase inhibitors for their effects on TNFα-mediated ICAM1 expression.
- To identify specific kinase inhibitors that induce adverse effects in kidney, heart, and liver cells.
- To correlate in vitro findings with clinical adverse events and validate a novel reporter system for KI safety assessment.
Main Methods:
- Screened 760 kinase inhibitors using hiPSC-derived renal proximal tubule-like cells (PTLCs), cardiomyocytes-like cells (CMLC), and hepatocyte-like cells (HLC) with an ICAM1-eGFP reporter.
- Analyzed kinase inhibitor activity data from the ChEMBL database to identify off-target effects.
- Correlated in vitro results with clinical data on renal adverse events associated with specific KIs.
Main Results:
- Four kinase inhibitors (Palbociclib, Miltefosine, Gilteritinib, Erdafitinib) increased ICAM1-eGFP reporter expression and chemokine release.
- Off-target kinase inhibition was associated with enhanced ICAM1-eGFP expression.
- KIs that increased ICAM1-eGFP expression in PTLCs were linked to renal adverse events in patients.
Conclusions:
- Imaging-based hiPSC-derived reporter cells offer a valuable tool for identifying kinase inhibitor liabilities.
- This approach can predict potential TNFα pathway modulation and associated organ toxicities.
- The study provides a framework for early detection of adverse effects of novel kinase inhibitors.
