Anidulafungin-Induced Suicidal Erythrocyte Death

Abstract

Background/Aims: The novel antifungal drug Anidulafungin is used for the treatment of diverse fungal infections including candidiasis and aspergillosis. The traditional antifungal drug amphotericin B has previously been shown to trigger eryptosis, the suicidal death of erythrocytes characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine translocation to the erythrocyte surface. Triggers of eryptosis include increase of cytosolic Ca²⁺ activity ([Ca²⁺]_i), oxidative stress, ceramide, activated protein kinase C (PKC), casein kinase 1α or p38 kinase and activated caspases. Inhibitors of eryptosis include nitric oxide (NO). The present study explored, whether Anidulafungin induces eryptosis. Methods: Flow cytometry was employed to estimate phosphatidylserine abundance at the erythrocyte surface from annexin-V-binding, cell volume from forward scatter, [Ca²⁺]_i from Fluo3-fluorescence, abundance of reactive oxygen species (ROS) from DCFDA dependent fluorescence, and ceramide abundance at the erythrocyte surface utilizing specific antibodies. Hemolysis was quantified by measuring haemoglobin concentration in the supernatant. Results: A 48 hours exposure of human erythrocytes to Anidulafungin (1.5 - 6 µg/ml) significantly increased hemolysis and the percentage of annexin-V-binding cells, and significantly decreased forward scatter. Anidulafungin (6 µg/ml) slightly, but significantly inceased Fluo3-fluorescence and the effect of Anidulafungin on annexin-V-binding was slightly, but significantly blunted by removal of extracellular Ca²⁺. The effect of Anidulafungin on annexin-V-binding was further significantly blunted by the p38 kinase inhibitor SB203580 (2 µM) and NO donor nitroprusside (1 µM). An increase of extracellular K⁺ concentration significantly blunted the effect of Anidulafungin on cell volume but not on annexin-V-binding. Anidulafungin rather decreased DCFDA fluorescence and the effect of Anidulafungin on annexin-V-binding was not significantly blunted by the antioxidant N-acetylcysteine (1 mM). Moreover, the effect of Anidulafungin on annexin-V-binding was not paralleled by significant increase of ceramide abundance and was not significantly blunted by PKC inhibitor staurosporine (1 µM), casein kinase 1α inhibitor D4476 (10 µM) or pancaspase inhibitor zVAD (10 µM). Conclusions: Anidulafungin triggers hemolysis and eryptosis with cell shrinkage and phospholipid scrambling of the erythrocyte cell membrane, an effect in part due to Ca²⁺ entry and activation of p38 kinase.

The Effects of Intravitreal Administration of Antifungal Drugs on the Structure and Mechanical Properties Peripheral Blood Erythrocyte Surface in Rabbits

“…Side effects of micafungin treatment are known to include microangiopathic hemolytic anemia and thrombocytopenia with microvascular thrombosis [ 5 ]. One possible mechanism of thrombosis development involves eryptosis, i.e., the suicidal death of erythrocytes characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine translocation to the erythrocyte surface [ 5 ]. Peter et al showed that micafungin triggers hemolysis and eryptosis with cell shrinkage and phospholipid scrambling of the erythrocyte cell membrane [ 5 ].…”

“…One possible mechanism of thrombosis development involves eryptosis, i.e., the suicidal death of erythrocytes characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine translocation to the erythrocyte surface [ 5 ]. Peter et al showed that micafungin triggers hemolysis and eryptosis with cell shrinkage and phospholipid scrambling of the erythrocyte cell membrane [ 5 ]. There is no evidence in the literature presenting similar behavior due to voriconazole intake.…”

“…Antifungal drugs introduced locally can act on blood cells, changing their properties, potentially contributing to the drugs’ negative side effects [ 5 ].…”

Suicidal death of erythrocytes in cancer and its chemotherapy: A potential target in the treatment of tumor‐associated anemia

“…Erythrocytes from newborns are highly sensitive to oxidative stress leading to rapid eryptosis and subsequent clearance of those erythrocytes following birth . Eryptosis is triggered by complement, hyperosmotic shock, energy depletion, oxidative stress, cellular K + loss, hyperthermia and a wide variety of xenobiotics including diverse cytostatic drugs (see below).…”

Triggering of Eryptosis, the Suicidal Erythrocyte Death by Mammalian Target of Rapamycin (mTOR) inhibitor Temsirolimus

“…Eryptosis is suppressed by AMP activated kinase AMPK [73], cGMP-dependent protein kinase [74], mitogen and stress activated kinase MSK1/2 [75], PAK2 kinase [76] and sorafenib/sunitinib sensitive kinases [77, 78]. Eryptosis is triggered by a wide variety of conditions including hyperosmotic shock [61], oxidative stress [61], energy depletion [61], or exposure to diverse xenobiotics [61, 79‒137]. Several clinical disorders are associated with enhanced eryptosis, such as iron deficiency [61], dehydration [138], hyperphosphatemia [139], chronic kidney disease (CKD) [140‒143], hemolytic-uremic syndrome [144], diabetes [145], hepatic failure [79], malignancy [146, 147], arteriitis [148], sepsis [149], sickle-cell disease [61], beta-thalassemia [61], Hb-C and G6PD-deficiency [61], Wilsons disease [149], as well as advanced age [150].…”