Deformability of blood cells is known to influence vascular flow and contribute to vascular complications. Medications for hematologic diseases have the potential to modulate these complications if they alter blood cell deformability. Here we report the effect of chemotherapy on leukemia cell mechanical properties. Acute lymphoblastic and acute myeloid leukemia cells were incubated with standard induction chemotherapy, and individual cell stiffness was tracked with atomic force microscopy. When exposed to dexamethasone or daunorubicin, leukemia cell stiffness increased by nearly 2 orders of magnitude, which decreased their passage through microfluidic channels. This stiffness increase occurred before caspase activation and peaked after completion of cell death, and the rate of stiffness increase depended on chemotherapy type. Stiffening with cell death occurred for all cell types investigated and may be due to dynamic changes in the actin cytoskeleton. These observations suggest that chemotherapy itself may increase the risk of vascular complications in acute leukemia.
IntroductionAlterations of biophysical properties of blood cells contribute to the pathophysiology of hematologic diseases. 1-3 While chemotherapy-induced cell death has been a mainstay of cancer treatment for decades and is well-studied biochemically, little is known about the mechanical effects chemotherapy may have on leukemia cells. Furthermore, since hyperleukocytosis accompanies some cases of acute leukemia, mechanical changes in leukemia cells due to chemotherapy could significantly alter the overall blood rheology.In this work, we quantified the effect of standard induction chemotherapy on the stiffness of acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML) cells using atomic force microscopy (AFM), a tool for imaging and characterization of materials at the nanometer scale. 4 The high force sensitivity of AFM and its ability to measure properties of individual cells over long times makes the technique particularly appropriate for measuring dynamic changes in cell stiffness. We find that when exposed to chemotherapy, leukemia cell stiffness increased by nearly 2 orders of magnitude at a rate dependent on the type of chemotherapy employed.
Materials and methods
Leukemia cell sources and reagentsLeukemia cells were isolated via centrifugation from the blood of patients with newly diagnosed acute leukemia noted to have peripheral blast cells. Leukemic cell lines were purchased commercially (ATCC, Manassas, VA). University of California, San Francisco (UCSF) and UC Berkeley institutional review boards approved all experimental procedures. Informed consent was obtained from each human subject, in accordance with the Declaration of Helsinki, before a blood sample was obtained. Dexamethasone and daunorubicin (Sigma-Aldrich, St Louis, MO) are mainstay induction chemotherapeutic agents for ALL and AML, respectively. Accordingly, lymphoid and myeloid leukemic cells were exposed to typical treatment doses of 1 M dexamethasone and 1 M...