Mitoxantrone-Induced Suicidal Erythrocyte Death

Abstract

Background/Aims: Mitoxantrone, a cytotoxic drug used for the treatment of malignancy and multiple sclerosis, is at least in part effective by triggering apoptosis. Similar to apoptosis of nucleated cells, erythrocytes may enter eryptosis, a type of suicidal cell death. Hallmarks of eryptosis are cell shrinkage and cell membrane scrambling with phosphatidylserine translocation to the erythrocyte surface. Signalling involved in eryptosis include Ca²⁺-entry, ceramide formation and oxidative stress. Methods: Cell volume was estimated from forward scatter, phosphatidylserine-exposure from annexin V binding, formation of reactive oxidant species (ROS) from 2′,7′-dichlorodihydrofluorescein-diacetate fluorescence, and ceramide abundance from binding of fluorescent antibodies in flow cytometry. Results: A 48 hours exposure to mitoxantrone was followed by significant decrease of forward scatter (≥ 5 μg/ml mitoxantrone) and increase of annexin-V-binding (≥ 10 μg/ml mitoxantrone), effects paralleled by significant increases of ROS formation (25 μg/ml mitoxantrone) and ceramide abundance (25 μg/ml mitoxantrone). The effect of mitoxantrone was not significantly modified by nominal absence of extracellular Ca²⁺ but significantly blunted by the antioxidant N-acetylcysteine (1 mM). Conclusions: Mitoxantrone triggers cell membrane scrambling, an effect not requiring entry of extracellular Ca²⁺ but at least partially due to formation of ROS and ceramide.

Stimulation of Suicidal Erythrocyte Death by PRIMA-1

“…PRIMA-1 tended to increase phosphatidylserine exposure at lower concentrations (10 µM), an effect, however, not reaching statistical significance. It must be kept in mind that erythrocytes may be sensitized to the effect of PRIMA-1 by other xenobiotics stimulating cell membrane scrambling [35,49,50,52,53,54,55,56,57,58,59,60,61,62,63,64,65,66,67,68,69,70,71,72,73,74,75,76,77,78,79,80,81,82] or by diseases associated with enhanced cell membrane scrambling, such as sepsis, malaria, sickle cell disease, Wilson's disease, iron deficiency, malignancy, metabolic syndrome, diabetes, hepatic failure, renal insufficiency, hemolytic uremic syndrome, hyperphosphatemia and phosphate depletion [34,83,84]. …”

Novel Approaches of Oxidative Stress Mechanisms in the Multiple Sclerosis Pathophysiology and Therapy

“…The study showed that mitoxantrone triggered cell apoptosis, partially due to the formation of ROS and ceramide, thus increasing OS. In addition, the authors assessed the effect of the antioxidant N-acetylcysteine, which significantly reduced the effect of mitoxantrone (105). Due to the fact that the studies are not conclusive, it appears that treatment with IFN-β and mitoxantrone does not reduce OS (103).…”

Therapeutic potential of manipulating suicidal erythrocyte death

“…Nevertheless, any of the small molecules listed in Table 2 could, at least in theory, accelerate the eryptosis of plasmodium-infected erythrocytes and thus counteract the rise of parasitemia. It is noteworthy, that the malaria pathogen plasmodium is not expected to become resistant to therapeutic acceleration of eryptosis, which is accomplished by cell mechanisms and is Estramustine 75 --100 µM + + [67] Ferutinin 30 µM + [68] Fluoxetine 25 --50 µM + [69] FTY720 10 µM + [1,2] Fumagillin 10 --100 µM + + [70] Gambogic acid 100 --500 nM + + [1,2] Gedunin 12 µM + [71] Geldanamycin 5 --50 µM + + [72] Glycation 40 mM glucose + Honokiol 5 --15 µM + + [74] Indoxyl sulfate 50 --600 µM + + Lumefantrine 3 --24 µg/ml [76] Lysophosphatidic acid 2.5 µM + + [77] Mercury 1 µM + + [78] Methyldopa 6 µg/ml + + Mitoxantrone 10 µg/ml + + [55] Monensin 0.1 µg/ml + [1,2] Mushroom tyrosinase 7 U/mL + + [81] Naphthoquinone derivatives 10 µM + [82] Nitazoxanide 1 --50 µg/ml + [83] Novobiocin 500 µM + + [84] Nystatin 5 --15 µg/ml + [85] Ochratoxin A 2.5 --10 µM + + Patulin 2.5 --10 µM + [87] Penta-O-galloyl-b-D-glucose 10 --50 µM + + [88] Peptidoglycan 10 µg/ml + [89] Phloretin 100 µM + [90] Phorbol-12 myristate-13 acetate…”