Nilotinib is a highly effective tyrosine kinase inhibitor used to treat Chronic Myeloid Leukemia (CML). While nilotinib successfully treats CML, a common and serious side effect is development of nilotinib-induced arterial disease (NAD) even in patients without pre-existing risk factors. This side effect is not seen with imatinib, a CML drug in the same class that inhibits the same target as nilotinib. To date, the mechanisms of NAD are unknown, and the cell types involved in this adverse effect have yet to be identified. We used a human induced pluripotent stem cell (hiPSC) model to study the effects of nilotinib on endothelial cells (hiPSC-ECs) and vascular smooth muscle cells (hiPSC-VSMCs) in vitro. In hiPSC-ECs, nilotinib adversely affects proliferation and migration, in addition to increasing intracellular nitric oxide. Nilotinib did not alter hiPSC-EC barrier function or lipoprotein uptake. There was no differential effect of nilotinib on hiPSC-VSMCs compared to imatinib across multiple assays including toxicity, migration, proliferation, and contraction. From these data we conclude that nilotinib-induced endothelial dysfunction is a key contributor to NAD, while it is less likely that VSMCs play a prominent role in this adverse effect. We next evaluated whether enhanced inhibition of the drug target ABL1 by nilotinib compared to imatinib explains the adverse effect of nilotinib on endothelial cells. CRISPR/Cas9 KO of ABL1 in multiple cell lines did not alter the nilotinib-specific endothelial phenotype, suggesting that the mechanism of nilotinib-induced endothelial dysfunction is independent of on-target inhibition of ABL1. To explore potential off-target mechanisms of NAD, we next conducted a pharmacologic screen to evaluate a subset of potential off-target candidates. We further studied our top two candidates from this screen, SRC and TEK, with a KO approach. Neither SRC nor TEK KO altered the effect of nilotinib on hiPSC-ECs, suggesting that these kinases are not central to the mechanism of nilotinib-induced endothelial dysfunction. Further research is needed to elucidate the mechanism of NAD. The tools and assays developed in the present study can be employed in future mechanistic and patient-specific pharmacogenomic studies to avert NAD and advance precision medicine for patients with CML.

Creator

• Pinheiro, Emily

DOI

• https://doi.org/10.21985/n2-bkfs-7v97

Subject

• Pharmacology

Language

• en

Alternate Identifier

• etdadmin_upload_1015403
• http://dissertations.umi.com/northwestern:16792

Keyword

• vascular smooth muscle cells
• nilotinib
• induced pluripotent stem cells
• CML
• endothelial cells

Date created

• 2023-01-01

Resource type

• Dissertation

Rights statement

• In Copyright