Oxidative stress monitoring in iPSC-derived motor neurons using genetically encoded biosensors of H2O2


Oxidative stress plays an important role in the development of neurodegenerative diseases, being either the initiator or part of a pathological cascade that leads to the neuron's death. Genetically encoded biosensors of oxidative stress demonstrated their general functionality and overall safety in various systems. However, there is still insufficient data regarding their use in the research of disease-related phenotypes in relevant model systems, such as human cells. Here, we establish an approach for monitoring the redox state of live motor neurons with SOD1 mutations associated with amyotrophic lateral sclerosis. Using CRISPR/Cas9, we insert genetically encoded biosensors of cytoplasmic and mitochondrial H2O2 in the genome of induced pluripotent stem cell (iPSC) lines. We demonstrate that the biosensors remain functional in motor neurons derived from these iPSCs and reflect the differences in the stationary redox state of the neurons with different genotypes. Moreover, we show that the biosensors respond to alterations in motor neuron oxidation caused by either environmental changes or cellular stress. Thus, the obtained platform is suitable for cell-based research of neurodegenerative mechanisms. © 2022. The Author(s).

Authors Ustyantseva E, Pavlova SV, Malakhova AA, Ustyantsev K, Zakian SM, Medvedev SP
Journal Scientific reports
Publication Date 2022 May 27;12(1):8928
PubMed 35624228
PubMed Central PMC9142597
DOI 10.1038/s41598-022-12807-z

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