TY - JOUR

T1 - Transgenic iPSC Lines with Genetically Encoded MitoTimer to Study Mitochondrial Biogenesis in Dopaminergic Neurons with Tauopathy

AU - Nadtochy, Julia A.

AU - Medvedev, Sergey P.

AU - Grigor’eva, Elena V.

AU - Pavlova, Sophia V.

AU - Minina, Julia M.

AU - Chechushkov, Anton V.

AU - Malakhova, Anastasia A.

AU - Kovalenko, Liudmila V.

AU - Zakian, Suren M.

N1 - The reported study was funded by the Foundation for Scientific and Technological Development of Yugra according to the research project No. 2023-573-05.

PY - 2025

Y1 - 2025

N2 - Background: Tauopathy has been identified as a prevalent causative agent of neurodegenerative diseases, including frontotemporal dementia with parkinsonism-17 (FTDP-17). This rare hereditary neurodegenerative condition is characterised by the manifestation of parkinsonism and behavioural changes. The majority of cases of FTDP-17 are associated with mutations in the MAPT gene, which encodes the tau protein. MAPT mutations lead to disruption of the balance between 3R and 4R tau forms, which causes destabilisation of microtubules and impairment of cellular organelle functions, particularly mitochondrial dysfunction. The development of model systems and tools for studying the molecular, genetic, and biochemical mechanisms underlying FTDP-17 and testing therapies at the cellular level is an urgent necessity. Methods: In this study, we generated transgenic lines of induced pluripotent stem cells (iPSCs) from a patient carrying the pathogenic mutation c.2013T > G (rs63750756, p.N279K) of MAPT and a healthy donor. A doxycycline-controlled transgene of the genetically encoded biosensor MitoTimer was integrated into the AAVS1 locus of these cells. The MitoTimer biosensor allows for lifetime monitoring of the turnover of mitochondria in neuronal cells derived from directed iPSC differentiation. The fact that transcription of the transgene can be induced by doxycycline provides additional possibilities for pulse labelling of newly formed mitochondria. Results: Transgenic iPSC lines provide a unique tool to study the molecular and genetic mechanisms of FTDP-17 caused by the presence of the c.2013T > G (p.N279K) mutation, as well as to test potential drugs in vitro.

AB - Background: Tauopathy has been identified as a prevalent causative agent of neurodegenerative diseases, including frontotemporal dementia with parkinsonism-17 (FTDP-17). This rare hereditary neurodegenerative condition is characterised by the manifestation of parkinsonism and behavioural changes. The majority of cases of FTDP-17 are associated with mutations in the MAPT gene, which encodes the tau protein. MAPT mutations lead to disruption of the balance between 3R and 4R tau forms, which causes destabilisation of microtubules and impairment of cellular organelle functions, particularly mitochondrial dysfunction. The development of model systems and tools for studying the molecular, genetic, and biochemical mechanisms underlying FTDP-17 and testing therapies at the cellular level is an urgent necessity. Methods: In this study, we generated transgenic lines of induced pluripotent stem cells (iPSCs) from a patient carrying the pathogenic mutation c.2013T > G (rs63750756, p.N279K) of MAPT and a healthy donor. A doxycycline-controlled transgene of the genetically encoded biosensor MitoTimer was integrated into the AAVS1 locus of these cells. The MitoTimer biosensor allows for lifetime monitoring of the turnover of mitochondria in neuronal cells derived from directed iPSC differentiation. The fact that transcription of the transgene can be induced by doxycycline provides additional possibilities for pulse labelling of newly formed mitochondria. Results: Transgenic iPSC lines provide a unique tool to study the molecular and genetic mechanisms of FTDP-17 caused by the presence of the c.2013T > G (p.N279K) mutation, as well as to test potential drugs in vitro.

KW - CRISPR/Cas9

KW - MAPT:c.2013T > G

KW - MitoTimer biosensor

KW - frontotemporal dementia with parkinsonism-17

KW - induced pluripotent stem cells

KW - mitochondrial dysfunction

UR - https://www.mendeley.com/catalogue/4f11c24d-113d-35e5-9190-8bd5c4c98f8b/

UR - https://www.scopus.com/record/display.uri?eid=2-s2.0-105000963942&origin=inward&txGid=0ef9b29c317fe41eb17b42e322bf2354

U2 - 10.3390/biomedicines13030550

DO - 10.3390/biomedicines13030550

M3 - Article

C2 - 40149527

VL - 13

JO - Biomedicines

JF - Biomedicines

SN - 2227-9059

IS - 3

M1 - 550

ER -