Dasatinib Reversibly Disrupts Endothelial Vascular Integrity by Increasing Non-Muscle Myosin II Contractility in a ROCK-Dependent Manner

Abstract

Purpose: Dasatinib is a short-acting dual ABL/SRC family tyrosine kinase inhibitor (TKI), which is frequently used to treat chronic myeloid leukemia. Although very effective, patients taking dasatinib often display severe adverse effects, including pleural effusions and increased risk of bleeding primarily in the gastrointestinal tract. The actual causes of these side effects are currently undetermined. We hypothesize that endothelial cells (ECs) that line the inner walls of blood vessels and control the traffic to the underlying tissues might be involved.Experimental Design: The effects of TKIs on ECs were studied by various assays, such as real-time cell impedance measurements, live-cell microscopy, wound healing, Western blot, and an in vivo model.Results: Dasatinib uniquely causes a profound, dose-dependent disorganization of the EC monolayers. Dasatinib promoted the disassembly of cell-cell contacts, altered cell-matrix contacts, and further altered the wound healing. A key observation is that this effect is fully reversible after drug washout. In line with these in vitro observations, intraperitoneal administration of dasatinib to mice caused significant vascular leakage in the intestine. The underlying molecular mechanism of dasatinib-induced reorganization of the actin involves ROCK activation, which increases the amount of the phosphorylation of myosin light chain and consequently activates the non-muscle myosin II.Conclusions: Our data are consistent with a scenario in which dasatinib triggers a transient increase in vascular leakage that probably contributes to adverse effects such as bleeding diathesis and pleural effusions.

A phosphoproteomic signature in endothelial cells predicts vascular toxicity of tyrosine kinase inhibitors used in CML

“…Imatinib was not associated with HUVEC toxicity in any assay. These data are consistent with most of the literature, although nilotinib did not enhance leukocyte adhesion to HUVECs, while it increased THP1 adhesion to EA.hy ECs, 10 and nilotinib did not impair wound healing in a prior study. 11 The variable effect of nilotinib may be due to a steep dose-response effect on ECs that also depends on EC type, 9,10 consistent with dose-dependent cardiovascular ischemia in nilotinib-treated CML patients.…”

“…Emerging data support that vasculotoxic TKIs directly damage ECs in vitro in a dose-dependent manner and enhance atherosclerosis in rodent models. [7][8][9] Specifically, dasatinib enhanced leukocyte adhesion molecule expression in human pulmonary ECs 7 and impaired wound healing in human umbilical vein endothelial cells (HUVECs), 10 nilotinib impaired proliferation and survival of human coronary ECs 9 and HUVECs, 11 and ponatinib impaired survival and migration and increased apoptosis in HUVECs. 11 Because these drugs are kinase inhibitors, we hypothesized that the vasculotoxicity of CML TKIs may be due to modulation of distinct signaling pathways in ECs when compared with nontoxic drugs, and if so, a phosphoproteomic profile could be identified to predict vascular toxicity of novel drugs.…”

<p>miR-31 Modulates Liver Cancer HepG2 Cell Apoptosis and Invasion via ROCK1/F-Actin Pathways</p>

“…Rho-associated coiled-coil containing protein kinase 1 (ROCK1)/F-actin pathway functions as a tumor suppressor in several types of cancer. 17,18 Activating ROCK1/F-actin pathway suppressed cell survival and migration in nonsmall cell lung cancer (NSCLC) A549 cells. 19 Moreover, increased evidence have revealed the relationship between mitochondrial and ROCK1/F-actin pathway.…”

Somatic Mutations in Clonally Expanded T-lymphocytes in Patients with Chronic Graft-Versus-Host Disease

“…Real-time electrical impedance measurements monitoring target cell killing have been widely applied to study the cellular cytotoxicity in vitro . ( 16, 17 ) To test the functional effects of the mutated CD4+ T cells and to verify aberrantly upregulated gene expression signatures associated with cytotoxicity, we performed co-culture experiments with CD4+ T cells and primary fibroblasts. Addition of purified CD4+ T cells on the mono-layer of primary fibroblasts from the index patient resulted in decreased electrical impedance implicating cytotoxicity of the CD4+ T cells (Figure 5A and B).…”