Coordinated time-dependent modulation of AMPK/Akt/mTOR signaling and autophagy controls osteogenic differentiation of human mesenchymal stem cells
Само за регистроване кориснике
2013
Аутори
Pantović, AleksandarKrstić, Aleksandra
Janjetović, Kristina
Krstić, Jelena
Harhaji-Trajković, Ljubica
Bugarski, Diana
Trajković, Vladimir
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
We investigated the role of AMP-activated protein kinase (AMPK), Akt, mammalian target of rapamycin (mTOR), autophagy and their interplay in osteogenic differentiation of human dental pulp mesenchymal stem cells. The activation of various members of AMPK, Akt and mTOR signaling pathways and autophagy was analyzed by immunoblotting, while osteogenic differentiation was assessed by alkaline phosphatase staining and real-time RT-PCR/immunoblot quantification of osteocalcin, Runt-related transcription factor 2 and bone morphogenetic protein 2 mRNA and/or protein levels. Osteogenic differentiation of mesenchymal stem cells was associated with early (day 1) activation of AMPK and its target Raptor, coinciding with the inhibition of mTOR and its substrate p70S6 kinase. The early induction of autophagy was demonstrated by accumulation of autophagosome-bound LC3-11, upregulation of proautophagic,beclin-1 and a decrease in the selective autophagic target p62. This was followed by the late activa...tion of Akt/mTOR at days 3-7 of differentiation. The RNA interference-mediated silencing of AMPK, mTOR or autophagy-essential LC3 beta, as well as the pharmacological inhibitors of AMPK (compound C), Akt (10-DEBC hydrochloride), mTOR (rapamycin) and autophagy (bafilomycin A1, chloroquine and ammonium chloride), each suppressed mesenchymal stem cell differentiation to osteoblasts. AMPK knockdown prevented early mTOR inhibition and autophagy induction, as well as late activation of Akt/mTOR signaling, while Ala inhibition suppressed mTOR activation without affecting AMPK phosphorylation. Our data indicate that AMPK controls osteogenic differentiation of human mesenchymal stem cells through both early mTOR inhibition-mediated autophagy and late activation of Akt/mTOR signaling axis.
Кључне речи:
Mesenchymal stem cells / Osteoblasts / AMPK / Akt / mTOR / AutophagyИзвор:
Bone, 2013, 52, 1, 524-531Издавач:
- Elsevier Science Inc, New York
Финансирање / пројекти:
- UNESCO L'OREAL National Scholarship Program "For Women in Science" [403F]
- Модулација сигналних путева који контролишу интрацелуларни енергетски баланс у терапији тумора и неуро-имуно-ендокриних поремећаја (RS-MESTD-Integrated and Interdisciplinary Research (IIR or III)-41025)
- Улога аутофагије у регулацији смрти туморских ћелија (RS-MESTD-Basic Research (BR or ON)-173053)
- Регенеративни и модулаторни потенцијал адултних матичних ћелија (RS-MESTD-Basic Research (BR or ON)-175062)
DOI: 10.1016/j.bone.2012.10.024
ISSN: 8756-3282
PubMed: 23111315
WoS: 000312750700062
Scopus: 2-s2.0-84870389278
Институција/група
Institut za medicinska istraživanjaTY - JOUR AU - Pantović, Aleksandar AU - Krstić, Aleksandra AU - Janjetović, Kristina AU - Krstić, Jelena AU - Harhaji-Trajković, Ljubica AU - Bugarski, Diana AU - Trajković, Vladimir PY - 2013 UR - http://rimi.imi.bg.ac.rs/handle/123456789/522 AB - We investigated the role of AMP-activated protein kinase (AMPK), Akt, mammalian target of rapamycin (mTOR), autophagy and their interplay in osteogenic differentiation of human dental pulp mesenchymal stem cells. The activation of various members of AMPK, Akt and mTOR signaling pathways and autophagy was analyzed by immunoblotting, while osteogenic differentiation was assessed by alkaline phosphatase staining and real-time RT-PCR/immunoblot quantification of osteocalcin, Runt-related transcription factor 2 and bone morphogenetic protein 2 mRNA and/or protein levels. Osteogenic differentiation of mesenchymal stem cells was associated with early (day 1) activation of AMPK and its target Raptor, coinciding with the inhibition of mTOR and its substrate p70S6 kinase. The early induction of autophagy was demonstrated by accumulation of autophagosome-bound LC3-11, upregulation of proautophagic,beclin-1 and a decrease in the selective autophagic target p62. This was followed by the late activation of Akt/mTOR at days 3-7 of differentiation. The RNA interference-mediated silencing of AMPK, mTOR or autophagy-essential LC3 beta, as well as the pharmacological inhibitors of AMPK (compound C), Akt (10-DEBC hydrochloride), mTOR (rapamycin) and autophagy (bafilomycin A1, chloroquine and ammonium chloride), each suppressed mesenchymal stem cell differentiation to osteoblasts. AMPK knockdown prevented early mTOR inhibition and autophagy induction, as well as late activation of Akt/mTOR signaling, while Ala inhibition suppressed mTOR activation without affecting AMPK phosphorylation. Our data indicate that AMPK controls osteogenic differentiation of human mesenchymal stem cells through both early mTOR inhibition-mediated autophagy and late activation of Akt/mTOR signaling axis. PB - Elsevier Science Inc, New York T2 - Bone T1 - Coordinated time-dependent modulation of AMPK/Akt/mTOR signaling and autophagy controls osteogenic differentiation of human mesenchymal stem cells EP - 531 IS - 1 SP - 524 VL - 52 DO - 10.1016/j.bone.2012.10.024 ER -
@article{ author = "Pantović, Aleksandar and Krstić, Aleksandra and Janjetović, Kristina and Krstić, Jelena and Harhaji-Trajković, Ljubica and Bugarski, Diana and Trajković, Vladimir", year = "2013", abstract = "We investigated the role of AMP-activated protein kinase (AMPK), Akt, mammalian target of rapamycin (mTOR), autophagy and their interplay in osteogenic differentiation of human dental pulp mesenchymal stem cells. The activation of various members of AMPK, Akt and mTOR signaling pathways and autophagy was analyzed by immunoblotting, while osteogenic differentiation was assessed by alkaline phosphatase staining and real-time RT-PCR/immunoblot quantification of osteocalcin, Runt-related transcription factor 2 and bone morphogenetic protein 2 mRNA and/or protein levels. Osteogenic differentiation of mesenchymal stem cells was associated with early (day 1) activation of AMPK and its target Raptor, coinciding with the inhibition of mTOR and its substrate p70S6 kinase. The early induction of autophagy was demonstrated by accumulation of autophagosome-bound LC3-11, upregulation of proautophagic,beclin-1 and a decrease in the selective autophagic target p62. This was followed by the late activation of Akt/mTOR at days 3-7 of differentiation. The RNA interference-mediated silencing of AMPK, mTOR or autophagy-essential LC3 beta, as well as the pharmacological inhibitors of AMPK (compound C), Akt (10-DEBC hydrochloride), mTOR (rapamycin) and autophagy (bafilomycin A1, chloroquine and ammonium chloride), each suppressed mesenchymal stem cell differentiation to osteoblasts. AMPK knockdown prevented early mTOR inhibition and autophagy induction, as well as late activation of Akt/mTOR signaling, while Ala inhibition suppressed mTOR activation without affecting AMPK phosphorylation. Our data indicate that AMPK controls osteogenic differentiation of human mesenchymal stem cells through both early mTOR inhibition-mediated autophagy and late activation of Akt/mTOR signaling axis.", publisher = "Elsevier Science Inc, New York", journal = "Bone", title = "Coordinated time-dependent modulation of AMPK/Akt/mTOR signaling and autophagy controls osteogenic differentiation of human mesenchymal stem cells", pages = "531-524", number = "1", volume = "52", doi = "10.1016/j.bone.2012.10.024" }
Pantović, A., Krstić, A., Janjetović, K., Krstić, J., Harhaji-Trajković, L., Bugarski, D.,& Trajković, V.. (2013). Coordinated time-dependent modulation of AMPK/Akt/mTOR signaling and autophagy controls osteogenic differentiation of human mesenchymal stem cells. in Bone Elsevier Science Inc, New York., 52(1), 524-531. https://doi.org/10.1016/j.bone.2012.10.024
Pantović A, Krstić A, Janjetović K, Krstić J, Harhaji-Trajković L, Bugarski D, Trajković V. Coordinated time-dependent modulation of AMPK/Akt/mTOR signaling and autophagy controls osteogenic differentiation of human mesenchymal stem cells. in Bone. 2013;52(1):524-531. doi:10.1016/j.bone.2012.10.024 .
Pantović, Aleksandar, Krstić, Aleksandra, Janjetović, Kristina, Krstić, Jelena, Harhaji-Trajković, Ljubica, Bugarski, Diana, Trajković, Vladimir, "Coordinated time-dependent modulation of AMPK/Akt/mTOR signaling and autophagy controls osteogenic differentiation of human mesenchymal stem cells" in Bone, 52, no. 1 (2013):524-531, https://doi.org/10.1016/j.bone.2012.10.024 . .