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

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

Journal

Scientific Reports
ISSN: 2045-2322

Output data

Year: 2022, Volume: 12, Number: 1, Article number : 8928, Pages count : DOI: 10.1038/s41598-022-12807-z

Authors

Ustyantseva Elizaveta ¹ , Pavlova Sophia V. ^1,2,3 , Malakhova Anastasia A. ^1,2,3 , Ustyantsev Kirill ¹ , Zakian Suren M. ^1,2,3 , Medvedev Sergey P. ^1,2,3

Affiliations

1	The Federal Research Center Institute of Cytology and Genetics, Siberian Branch of Russian Academy of Sciences, 10, Lavrentiev Ave, 630090 Novosibirsk, Russia
2	Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, 8, Lavrentiev Ave., 630090 Novosibirsk, Russia
3	3 E. Meshalkin National Medical Research Center of the Ministry of Health of the Russian Federation, 15 Rechkunovskaya Str., 630055 Novosibirsk, Russia

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).

Ustyantseva E. , Pavlova S.V. , Malakhova A.A. , Ustyantsev K. , Zakian S.M. , Medvedev S.P.
Oxidative stress monitoring in iPSC-derived motor neurons using genetically encoded biosensors of H2O2
Scientific Reports. 2022. V.12. N1. 8928 . DOI: 10.1038/s41598-022-12807-z WOS Scopus OpenAlex

Web of science:	WOS:000800769400023
Scopus:	2-s2.0-85130770560
OpenAlex:	W4281876015

DB	Citing
Scopus	5
Web of science	5
OpenAlex	7