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Updated: Feb 22, 2026
Induction of Eryptosis in Red Blood Cells Using a Calcium Ionophore
Published on: January 21, 2020
Inhibition of Erythrocyte Cell Membrane Scrambling by ASP3026
A Al Mamun Bhuyan1, Rosi Bissinger1, Hang Cao1
1Department of Internal Medicine III, Eberhard-Karls-University of Tuebingen, Tuebingen, Germany.
Background/Aims:
The anaplastic lymphoma kinase (ALK) inhibitor ASP3026 is in clinical development for the treatment of ALK expressing non-small cell lung carcinoma (NSCLC). ASP3026 is in part effective by inducing apoptosis of tumor cells. Erythrocytes lack mitochondria and nuclei, key organelles in the execution of apoptosis, but are nevertheless able to enter suicidal death or eryptosis, which is characterized by cell membrane scrambling with phosphatidylserine translocation to the cell surface and by cell shrinkage. Eryptosis is triggered by cell stress, such as energy depletion, hyperosmotic shock, oxidative stress and excessive increase of cytosolic Ca2+ activity ([Ca2+]i). The present study explored, whether ASP3026 impacts on eryptosis.
Methods:
Human erythrocytes have been exposed to energy depletion (glucose withdrawal for 48 hours), oxidative stress (addition of 0.3 mM tert-butylhydroperoxide [tBOOH] for 50 min) or Ca2+ loading with Ca2+ ionophore ionomycin (1 µM for 60 min) in absence and presence of ASP3026 (1-4 µg/ml). Flow cytometry was employed to quantify phosphatidylserine exposure at the cell surface from annexin-V-binding, and cell volume from forward scatter.
Results:
Treatment with ASP3026 alone did not significantly modify annexin-V-binding or forward scatter. Energy depletion, oxidative stress and ionomycin, all markedly and significantly increased the percentage of annexin-V-binding erythrocytes, and decreased the forward scatter. ASP3026 significantly blunted the effect of energy depletion and oxidative stress, but not of ionomycin on annexin-V-binding. ASP3026 did not significantly influence the effect of any maneuver on forward scatter.
Conclusions:
ASP3026 is a novel inhibitor of erythrocyte cell membrane scrambling following energy depletion and oxidative stress.
Insights
The anaplastic lymphoma kinase (ALK) inhibitor ASP3026 prevents suicidal erythrocyte death (eryptosis) triggered by energy depletion and oxidative stress. This finding suggests a novel mechanism beyond its anti-cancer effects in non-small cell lung carcinoma.
Area of Science:
- Biochemistry
- Cell Biology
- Pharmacology
Background:
- Anaplastic lymphoma kinase (ALK) inhibitor ASP3026 targets ALK-expressing non-small cell lung carcinoma (NSCLC).
- Tumor cell apoptosis is a key mechanism of ASP3026 action.
- Erythrocytes undergo eryptosis, a form of suicidal cell death, under stress conditions, characterized by phosphatidylserine exposure and cell shrinkage.
Purpose of the Study:
- To investigate the impact of ASP3026 on eryptosis in human erythrocytes.
- To determine if ASP3026 modulates eryptosis induced by specific cellular stressors.
Main Methods:
- Human erythrocytes were subjected to energy depletion, oxidative stress, or calcium loading.
- Cells were treated with varying concentrations of ASP3026.
- Flow cytometry was used to quantify phosphatidylserine exposure (annexin-V binding) and cell volume (forward scatter).
Main Results:
- ASP3026 alone did not affect erythrocyte membrane scrambling or cell volume.
- Energy depletion, oxidative stress, and ionomycin significantly induced eryptosis markers.
- ASP3026 significantly inhibited phosphatidylserine exposure induced by energy depletion and oxidative stress, but not by ionomycin.
Conclusions:
- ASP3026 acts as a novel inhibitor of erythrocyte membrane scrambling.
- The drug's effect is specific to eryptosis induced by energy depletion and oxidative stress.
- This suggests a potential for ASP3026 to influence erythrocyte health beyond its anti-cancer role.
