PMID- 26143260 OWN - NLM STAT- MEDLINE DCOM- 20171222 LR - 20211204 IS - 1559-1182 (Electronic) IS - 0893-7648 (Linking) VI - 53 IP - 6 DP - 2016 Aug TI - S6K Promotes Dopaminergic Neuronal Differentiation Through PI3K/Akt/mTOR-Dependent Signaling Pathways in Human Neural Stem Cells. PG - 3771-3782 LID - 10.1007/s12035-015-9325-9 [doi] AB - It has recently been reported that the phosphoinositide 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) signaling pathway regulates neuronal differentiation of neural stem cells (NSCs) derived from rats or mice and is essential for the self-renewal of human embryonic stem cells (hESCs). However, the roles of PI3K/Akt/mTOR signaling pathways during proliferation and dopaminergic neuronal differentiation of human neural stem cells (hNSCs) are poorly understood. In this study, we examined the effect of regulation of these intracellular signaling pathways in hNSCs on the potential to maintain proliferation and induce dopaminergic neuronal differentiation. Dopaminergic neuronal differentiation depended on the concentration of insulin in our culture system. Inhibition of PI3K/Akt with LY294002 reduced proliferation and inhibited dopaminergic neuronal differentiation of these cells. We also found that rapamycin, a specific inhibitor of mTOR, significantly reduced neuronal differentiation without affecting proliferation. Inhibition of the Akt/mTOR signaling pathway led to inhibition of p70 ribosomal S6 kinase (S6K) signaling, which reduced dopaminergic neuronal differentiation in hNSCs. Inhibition of S6K by a specific chemical inhibitor, PF-4708671 inhibited dopaminergic neuronal differentiation of hNSCs. As expected, transduction with a dominant negative S6K1 (S6K1-DN) construct impaired dopaminergic neuronal differentiation of hNSCs. Conversely, overexpression of constitutively active S6K1 (S6K1-CA) promoted dopaminergic neuronal differentiation of these cells. In a survival study, 4 weeks after transplantation, no or very few donor cells were viable in striata grafted with S6K1-DN-transduced hNSCs. In contrast, S6K1-CA-transduced hNSCs survived, integrated into striata to generate tubular masses of grafts and differentiated toward TH-positive cells. Taken together, these data demonstrated that insulin promotes dopaminergic neuronal differentiation through a PI3K/Akt/mTOR-dependent pathway and that S6K plays a critical role in dopaminergic neuronal differentiation in hNSCs. FAU - Lee, Jeong Eun AU - Lee JE AD - Hanyang Biomedical Research Institute, Hanyang University, 133-791, Seoul, Republic of Korea. AD - Department of Pharmacology, College of Medicine, Hanyang University, 133-791, Seoul, Republic of Korea. FAU - Lim, Mi Sun AU - Lim MS AD - Hanyang Biomedical Research Institute, Hanyang University, 133-791, Seoul, Republic of Korea. AD - Graduate School of Biomedical Science and Engineering, Hanyang University, 133-791, Seoul, Republic of Korea. FAU - Park, Jae Hyun AU - Park JH AD - Hanyang Biomedical Research Institute, Hanyang University, 133-791, Seoul, Republic of Korea. AD - Department of Pharmacology, College of Medicine, Hanyang University, 133-791, Seoul, Republic of Korea. FAU - Park, Chang Hwan AU - Park CH AD - Hanyang Biomedical Research Institute, Hanyang University, 133-791, Seoul, Republic of Korea. chshpark@hanyang.ac.kr. AD - Graduate School of Biomedical Science and Engineering, Hanyang University, 133-791, Seoul, Republic of Korea. chshpark@hanyang.ac.kr. FAU - Koh, Hyun Chul AU - Koh HC AD - Hanyang Biomedical Research Institute, Hanyang University, 133-791, Seoul, Republic of Korea. hckoh@hanyang.ac.kr. AD - Department of Pharmacology, College of Medicine, Hanyang University, 133-791, Seoul, Republic of Korea. hckoh@hanyang.ac.kr. LA - eng PT - Journal Article DEP - 20150705 PL - United States TA - Mol Neurobiol JT - Molecular neurobiology JID - 8900963 RN - 0 (Insulin) RN - EC 1.14.16.2 (Tyrosine 3-Monooxygenase) RN - EC 2.7.11.1 (Proto-Oncogene Proteins c-akt) RN - EC 2.7.11.1 (Ribosomal Protein S6 Kinases, 70-kDa) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) SB - IM MH - Animals MH - Cell Count MH - Cell Differentiation/*drug effects MH - Cell Lineage/drug effects MH - Cell Proliferation/drug effects MH - Cell Survival/drug effects MH - Cells, Cultured MH - Dopaminergic Neurons/*cytology/drug effects/metabolism MH - Human Embryonic Stem Cells/cytology/drug effects/metabolism MH - Humans MH - Insulin/pharmacology MH - Neural Stem Cells/cytology/drug effects/*metabolism MH - Phosphatidylinositol 3-Kinases/*metabolism MH - Proto-Oncogene Proteins c-akt/*metabolism MH - Rats MH - Ribosomal Protein S6 Kinases, 70-kDa/*metabolism MH - Signal Transduction/drug effects MH - Stem Cell Transplantation MH - TOR Serine-Threonine Kinases/*metabolism MH - Transduction, Genetic MH - Tyrosine 3-Monooxygenase/metabolism OTO - NOTNLM OT - Dopaminergic neuronal differentiation OT - Human neural stem cells OT - PI3K/Akt/mTOR signaling OT - Proliferation OT - S6K EDAT- 2015/07/06 06:00 MHDA- 2017/12/23 06:00 CRDT- 2015/07/06 06:00 PHST- 2015/01/27 00:00 [received] PHST- 2015/06/25 00:00 [accepted] PHST- 2015/07/06 06:00 [entrez] PHST- 2015/07/06 06:00 [pubmed] PHST- 2017/12/23 06:00 [medline] AID - 10.1007/s12035-015-9325-9 [pii] AID - 10.1007/s12035-015-9325-9 [doi] PST - ppublish SO - Mol Neurobiol. 2016 Aug;53(6):3771-3782. doi: 10.1007/s12035-015-9325-9. Epub 2015 Jul 5.