Modeling ferroptosis in human dopaminergic neurons: Pitfalls and opportunities for neurodegeneration research
Nadine Renner,
Franziska Schöb,
Regina Pape,
Ilinca Suciu,
Anna-Sophie Spreng,
Anna-Katharina Ückert,
Eike Cöllen,
Federica Bovio,
Bruno Chilian,
Johannes Bauer,
Stefan Röpcke,
Jörg Bergemann,
Marcel Leist,
Stefan Schildknecht
Affiliations
Nadine Renner
Albstadt-Sigmaringen University, Faculty of Life Sciences, 72488, Sigmaringen, Germany
Franziska Schöb
In Vitro Toxicology and Biomedicine, Department of Biology, University of Konstanz, 78457, Konstanz, Germany
Regina Pape
In Vitro Toxicology and Biomedicine, Department of Biology, University of Konstanz, 78457, Konstanz, Germany
Ilinca Suciu
In Vitro Toxicology and Biomedicine, Department of Biology, University of Konstanz, 78457, Konstanz, Germany
Anna-Sophie Spreng
In Vitro Toxicology and Biomedicine, Department of Biology, University of Konstanz, 78457, Konstanz, Germany
Anna-Katharina Ückert
In Vitro Toxicology and Biomedicine, Department of Biology, University of Konstanz, 78457, Konstanz, Germany
Eike Cöllen
In Vitro Toxicology and Biomedicine, Department of Biology, University of Konstanz, 78457, Konstanz, Germany
Federica Bovio
Department of Biotechnology and Biosciences, University of Milano-Bicocca, 20126, Milano, Italy
Bruno Chilian
TRI Thinking Research Instruments GmbH, Große Freiheit 77, 22767, Hamburg, Germany
Johannes Bauer
TRI Thinking Research Instruments GmbH, Große Freiheit 77, 22767, Hamburg, Germany
Stefan Röpcke
Stemick GmbH, Byk-Gulden Str. 2, 78467, Konstanz, Germany
Jörg Bergemann
Albstadt-Sigmaringen University, Faculty of Life Sciences, 72488, Sigmaringen, Germany
Marcel Leist
In Vitro Toxicology and Biomedicine, Department of Biology, University of Konstanz, 78457, Konstanz, Germany
Stefan Schildknecht
Albstadt-Sigmaringen University, Faculty of Life Sciences, 72488, Sigmaringen, Germany; Corresponding author. Albstadt-Sigmaringen University, Faculty of Life Sciences, Anton Guenther Str. 51, 72488, Sigmaringen, Germany.
The activation of ferroptosis is being pursued in cancer research as a strategy to target apoptosis-resistant cells. By contrast, in various diseases that affect the cardiovascular system, kidneys, liver, and central and peripheral nervous systems, attention is directed toward interventions that prevent ferroptotic cell death. Mechanistic insights into both research areas stem largely from studies using cellular in vitro models. However, intervention strategies that show promise in cellular test systems often fail in clinical trials, which raises concerns regarding the predictive validity of the utilized in vitro models.In this study, the human LUHMES cell line, which serves as a model for human dopaminergic neurons, was used to characterize factors influencing the activation of ferroptosis. Erastin and RSL-3 induced cell death that was distinct from apoptosis. Parameters such as the differentiation state of LUHMES cells, cell density, and the number and timing of medium changes were identified as determinants of sensitivity to ferroptosis activation. In differentiated LUHMES cells, interventions at mechanistically divergent sites (iron chelation, coenzyme Q10, peroxidase mimics, or inhibition of 12/15-lipoxygenase) provide almost complete protection from ferroptosis. LUHMES cells allowed the experimental modulation of intracellular iron concentrations and demonstrated a correlation between intracellular iron levels, the rate of lipid peroxidation, as well as the sensitivity of the cells to ferroptotic cell death.These findings underscore the importance of understanding the various factors that influence ferroptosis activation and highlight the need for well-characterized in vitro models to enhance the reliability and predictive value of observations in ferroptosis research, particularly when translating findings into in vivo contexts.
