Detection of ER Stress in iPSC-Derived Neurons Carrying the p.N370S Mutation in the GBA1 Gene. / Yarkova, Elena S.; Grigor’eva, Elena V.; Medvedev, Sergey P. et al.
In: Biomedicines, Vol. 12, No. 4, 774, 04.2024.Research output: Contribution to journal › Article › peer-review
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TY - JOUR
T1 - Detection of ER Stress in iPSC-Derived Neurons Carrying the p.N370S Mutation in the GBA1 Gene
AU - Yarkova, Elena S.
AU - Grigor’eva, Elena V.
AU - Medvedev, Sergey P.
AU - Tarasevich, Denis A.
AU - Pavlova, Sophia V.
AU - Valetdinova, Kamila R.
AU - Minina, Julia M.
AU - Zakian, Suren M.
AU - Malakhova, Anastasia A.
N1 - This research was funded by Russian Science Foundation, grant number 23-15-00224 (https://rscf.ru/en/project/23-15-00224/, accessed on 21 March 2024).
PY - 2024/4
Y1 - 2024/4
N2 - Endoplasmic reticulum (ER) stress is involved in the pathogenesis of many human diseases, such as cancer, type 2 diabetes, kidney disease, atherosclerosis and neurodegenerative diseases, in particular Parkinson’s disease (PD). Since there is currently no treatment for PD, a better understanding of the molecular mechanisms underlying its pathogenesis, including the mechanisms of the switch from adaptation in the form of unfolded protein response (UPR) to apoptosis under ER stress conditions, may help in the search for treatment methods. Genetically encoded biosensors based on fluorescent proteins are suitable tools that facilitate the study of living cells and visualization of molecular events in real time. The combination of technologies to generate patient-specific iPSC lines and genetically encoded biosensors allows the creation of cell models with new properties. Using CRISPR-Cas9-mediated homologous recombination at the AAVS1 locus of iPSC with the genetic variant p.N370S (rs76763715) in the GBA1 gene, we created a cell model designed to study the activation conditions of the IRE1-XBP1 cascade of the UPR system. The cell lines obtained have a doxycycline-dependent expression of the genetically encoded biosensor XBP1-TagRFP, possess all the properties of human pluripotent cells, and can be used to test physical conditions and chemical compounds that affect the development of ER stress, the functioning of the UPR system, and in particular, the IRE1-XBP1 cascade.
AB - Endoplasmic reticulum (ER) stress is involved in the pathogenesis of many human diseases, such as cancer, type 2 diabetes, kidney disease, atherosclerosis and neurodegenerative diseases, in particular Parkinson’s disease (PD). Since there is currently no treatment for PD, a better understanding of the molecular mechanisms underlying its pathogenesis, including the mechanisms of the switch from adaptation in the form of unfolded protein response (UPR) to apoptosis under ER stress conditions, may help in the search for treatment methods. Genetically encoded biosensors based on fluorescent proteins are suitable tools that facilitate the study of living cells and visualization of molecular events in real time. The combination of technologies to generate patient-specific iPSC lines and genetically encoded biosensors allows the creation of cell models with new properties. Using CRISPR-Cas9-mediated homologous recombination at the AAVS1 locus of iPSC with the genetic variant p.N370S (rs76763715) in the GBA1 gene, we created a cell model designed to study the activation conditions of the IRE1-XBP1 cascade of the UPR system. The cell lines obtained have a doxycycline-dependent expression of the genetically encoded biosensor XBP1-TagRFP, possess all the properties of human pluripotent cells, and can be used to test physical conditions and chemical compounds that affect the development of ER stress, the functioning of the UPR system, and in particular, the IRE1-XBP1 cascade.
KW - CRISPR/Cas9
KW - ER stress
KW - GBA1
KW - Parkinson’s disease
KW - biosensors
KW - endoplasmic reticulum
KW - induced pluripotent stem cells
UR - https://www.scopus.com/record/display.uri?eid=2-s2.0-85191749373&origin=inward&txGid=adcd9f190dbcca071fe73b12f6e6570b
UR - https://www.mendeley.com/catalogue/3c5bf3fa-6265-3f7f-8798-5c55b4690089/
U2 - 10.3390/biomedicines12040744
DO - 10.3390/biomedicines12040744
M3 - Article
C2 - 38672099
VL - 12
JO - Biomedicines
JF - Biomedicines
SN - 2227-9059
IS - 4
M1 - 774
ER -
ID: 61084472