AUTHOR=de la Cruz Alicia , Vera-Zambrano Alba , Peraza Diego A. , Valenzuela Carmen , Zapata Juan M. , Perez-Chacon Gema , Gonzalez Teresa 

TITLE=Fludarabine Inhibits KV1.3 Currents in Human B Lymphocytes

JOURNAL=Frontiers in Pharmacology

VOLUME=8

YEAR=2017

URL=https://www.frontiersin.org/journals/pharmacology/articles/10.3389/fphar.2017.00177

DOI=10.3389/fphar.2017.00177

ISSN=1663-9812

ABSTRACT=<p>Fludarabine (F-ara-A) is a purine analog commonly used in the treatment of indolent B cell malignancies that interferes with different aspects of DNA and RNA synthesis. K<sub>V</sub>1.3 K<sup>+</sup> channels are membrane proteins involved in the maintenance of K<sup>+</sup> homeostasis and the resting potential of the cell, thus controlling signaling events, proliferation and apoptosis in lymphocytes. Here we show that F-ara-A inhibits K<sub>V</sub> currents in human B lymphocytes. Our data indicate that K<sub>V</sub>1.3 is expressed in both BL2 and Dana B cell lines, although total K<sub>V</sub>1.3 levels were higher in BL2 than in Dana cells. However, K<sub>V</sub> currents in the plasma membrane were similar in both cell lines and were abrogated by the specific K<sub>V</sub>1.3 channel inhibitor PAP-1, indicating that K<sub>V</sub>1.3 accounts for most of the K<sub>V</sub> currents in these cell lines. F-ara-A, at a concentration (3.5 μM) similar to that achieved in the plasma of fludarabine phosphate-treated patients (3 μM), inhibited K<sub>V</sub>1.3 currents by 61 ± 6.3% and 52.3 ± 6.3% in BL2 and Dana B cells, respectively. The inhibitory effect of F-ara-A was concentration-dependent and showed an <italic>IC<sub>50</sub></italic> value of 0.36 ± 0.04 μM and a <italic>n<sub>H</sub></italic> value of 1.07 ± 0.15 in BL2 cells and 0.34 ± 0.13 μM (<italic>IC<sub>50</sub></italic>) and 0.77 ± 0.11 (<italic>n<sub>H</sub></italic>) in Dana cells. F-ara-A inhibition of plasma membrane K<sub>V</sub>1.3 was observed irrespective of its cytotoxic effect on the cells, BL2 cells being sensitive and Dana cells resistant to F-ara-A cytotoxicity. Interestingly, PAP-1, at concentrations as high as 10 μM, did not affect the viability of BL2 and Dana cells, indicating that blockage of K<sub>V</sub>1.3 in these cells is not toxic. Finally, F-ara-A had no effect on ectopically expressed K<sub>V</sub>1.3 channels, suggesting an indirect mechanism of current inhibition. In summary, our results describe the inhibitory effect of F-ara-A on the activity of K<sub>V</sub>1.3 channel. Although K<sub>V</sub>1.3 inhibition is not sufficient to induce cell death, further research is needed to determine whether it might still contribute to F-ara-A cytotoxicity in sensitive cells or be accountable for some of the clinical side effects of the drug.</p>