G protein accelerates epidermal cell migration via activation of PI3K/Akt-dependent mTORC1 signaling under hypoxia
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA549667 | ENA |
REPOSITORIES: ENA
Similar Datasets
Project description:When skin tissue is damaged, the wound microenvironment is hypoxic or anoxic due to the rupture of blood vessels and the high oxygen consumption of related cells; so far, it is not clear whether such hypoxic microenvironment will promote epidermal cell migration and the underlying molecular regulatory mechanisms of this effect remains unclear. Studies to date have shown that, immortal keratinocyte cell line HaCaT and primary human keratinocytes are maintained under conditions of hypoxia (1% oxygen) or normoxia. The researchers would adopt methods such as live cell imaging system, Western blotting, transwell assays and wound scratch assays etc. to study the changes of cell migration. Expression profile of mRNAs in HaCaT cells from 3 hypoxia and 3 normoxic conditions were analyzed by the Clariom D microarray assay. Gene Ontology (GO) and KEGG genomics analyses were performed to identify significant functions, pathways, and the associations of differentially expressed mRNAs. Moreover, the potential mechanism is discussed and studied. According to relevant results, the epidermal cell migration is promoted in the early hypoxia. Furthermore, experiment showed that 1456 mRNAs showed differential expression between the hypoxia and normoxic conditions, which included 537 upregulated and 919 downregulated mRNAs. These results also shown that all G proteins are upregulated. At the same time, GO analysis indicated that most upregulated mRNA are in connection with cell inflammation and migration. Pathway analysis indicated that 20 pathways corresponded to the upregulated mRNA and 6 pathways corresponded to the upregulated mRNA. These pathway analyses have indicated that genes involved in inflammation, migration and PI3K-Akt signaling are potentially regulated. Combine with microarray assay, we further shown that G protein accelerates epidermal cell migration via activation of PI3K/Akt-dependent mTORC1 signaling under the condition of hypoxia. Based on this evidence, researchers can further reveal the molecular and cellular mechanisms of local wound hypoxia, and for improving the wound healing.
2019-06-20 | GSE132988 | GEO
Project description:Dual function of the PI3K-Akt-mTORC1 axis in myelination of the peripheral nervous system
| PRJEB20661 | ENA
Project description:The AKT-mTOR pathway is a central regulator of cell growth and metabolism. Upon sustained mTOR activity, AKT activity is attenuated by a feedback loop that restrains upstream signaling. However, how cells control the signals that limit AKT activity is not fully understood. Here we show that MASTL/Greatwall, a cell-cycle kinase that supports mitosis by phosphorylating the PP2A/B55 inhibitors ENSA/ARPP19, inhibits PI3K-AKT activity by sustaining mTORC1- and S6K1-dependent phosphorylation of IRS1 and GRB10. Genetic depletion of MASTL results in an inefficient feedback loop and AKT hyperactivity. These defects are rescued by expression of phospho-mimetic ENSA/ARPP19 or inhibition of PP2A/B55 phosphatases. MASTL is directly phosphorylated by mTORC1, thereby limiting the PP2A/B55-dependent dephosphorylation of IRS1 and GRB10 downstream of mTORC1. Downregulation of MASTL results in increased glucose uptake in vitro and increased glucose tolerance in adult mice, suggesting the relevance of the MASTL-PP2A/B55 kinase-phosphatase module in controlling AKT and maintaining metabolic homeostasis.
2022-11-23 | PXD032796 | Pride
Project description:Hyperactivation of the phosphatydil-inositol-3' phosphate kinase (PI3K)/AKT pathway is observed in most NSCLCs, promoting proliferation, migration, invasion and resistance to therapy. AKT can be activated through several mechanisms that include loss of the negative regulator PTEN, activating mutations of the catalytic subunit of PI3K (PIK3CA) and/or mutations of AKT1 itself. However, number and identity of downstream targets of activated PI3K/AKT pathway are poorly defined. To identify the genes that are targets of constitutive PI3K/AKT signalling in lung cancer cells, we performed a comparative transcriptomic analysis of human lung epithelial cells (BEAS-2B) expressing active mutant AKT1 (AKT1-E17K), active mutant PIK3CA (PIK3CA-E545K) or that are silenced for PTEN. For each sample, 500 ng of total RNA were used to synthesize biotinylated cRNA with Illumina RNA Amplification Kit (Ambion, Austin, TX). Synthesis was carried out according to the manufacturersâ instructions. From each sample, technical triplicates were produced and 750 ng cRNA were hybridized for 18h to Human HT-12_V3_0_R1 Expression BeadChips (Illumina, San Diego, CA). Hybridized chips were washed and stained with streptavidin-conjugated Cy3 (GE Healthcare, Milan, Italy). BeadChips were dried and scanned with an Illumina Bead Array Reader (Illumina).
2016-12-01 | E-MTAB-5286 | biostudies-arrayexpress
Project description:This SuperSeries is composed of the following subset Series: GSE35701: CP001: Modulation of glutamine metabolism by the PI3K-PKB/c-akt-FOXO network regulates autophagy GSE35703: CP003: Modulation of glutamine metabolism by the PI3K-PKB/c-akt-FOXO network regulates autophagy Refer to individual Series
2012-07-21 | E-GEOD-35705 | biostudies-arrayexpress
Project description:Neuroblastoma is a pediatric tumor of the peripheral sympathetic nervous system with a highly variable prognosis. Activation of the PI3K/AKT pathway in neuroblastoma is correlated with poor patient prognosis, but the precise downstream effectors mediating this effect have not been determined. Here, we identify the forkhead transcription factor FOXO3a as a key target of the PI3K/AKT pathway in neuroblastoma. FOXO3a expression was elevated in low stage neuroblastoma tumors and normal embryonal neuroblasts, but reduced in late stage neuroblastoma. Inactivation of FOXO3a by AKT was essential for neuroblastoma cell survival. Treatment of neuroblastoma cells with the dual PI3K/mTOR inhibitor PI-103 activated FOXO3a and triggered apoptosis. This effect was rescued by FOXO3a silencing. Conversely, apoptosis induced by PI-103 or the AKT inhibitor MK-2206 was potentiated by FOXO3a overexpression. Further, levels of total or phosphorylated FOXO3a correlated closely with apoptotic sensitivity to MK-2206. In clinical specimens, there was an inverse relationship between gene expression signatures regulated by PI3K signaling and FOXO3a transcriptional activity. Moreover, high PI3K activity and low FOXO3a activity were each associated with an extremely poor prognosis. Our work indicates that expression of FOXO3a and its targets offer useful prognostic markers as well as biomarkers for PI3K/AKT inhibitor efficacy in neuroblastoma. Affymetrix U133 Plus 2.0 profiling of SY5Y-TetR-FOXO3A cells treated with doxycycline and/or the PI3K/mTOR inhibitor PI-103. Each condition profiled in triplicate.
2013-05-01 | E-GEOD-42762 | biostudies-arrayexpress
Project description:Caulis Polygoni Multiflori accelerates megakaryopoiesis and thrombopoiesis via activating PI3K/Akt and MEK/ERK (MAPK) signaling pathways
| PRJNA874815 | ENA
Project description:MiT/TFE transcriptional activity controls lysosomal biogenesis and is negatively regulated by the nutrient sensor mTORC1. Some tumors bypass this regulatory circuit via genetic alterations that drive MiT/TFE expression and activity; however, the mechanisms by which cells with intact or constitutive mTORC1 signaling maintain lysosomal catabolism remain to be elucidated. Using the murine epidermis as a model system, we find that epidermal Tsc1 deletion results in a wavy hair phenotype due to increased EGFR degradation. Unexpectedly, constitutive mTORC1 activation increases lysosomal content via up-regulated expression and activity of MiT/TFEs, while genetic or prolonged pharmacologic mTORC1 inactivation has the reverse effect. This paradoxical up-regulation of lysosomal biogenesis by mTORC1 is mediated by feedback inhibition of AKT, and a resulting suppression of AKT-induced MiT/TFE proteasomal degradation. These data suggest that oncogenic feedback loops work to restrain or maintain cellular lysosomal content during chronically inhibited or constitutively active mTORC1 signaling respectively, and reveal a mechanism by which mTORC1 regulates upstream receptor tyrosine kinase signaling.
2020-04-14 | GSE124754 | GEO
Project description:Sarcomas encompass heterogenous, difficult to treat cancers, lacking common therapeutic targets. Phosphatidylinositol-3 kinase (PI3K) signaling is activated in sarcomas to a greater degree than previously appreciated due to phosphatase and tensin homolog (PTEN) loss, and could represent such a target. Targeting PI3K signaling has largely focused on targeting mTORC1, considered the main effector of PI3K signaling, but this has not translated to success in the clinic, suggesting that there may be other effectors downstream of PI3K. One gap in our understanding of the PI3K signaling pathway has been the absence of a known oncogenic transcription factor. Herein we implicate TAZ and YAP as additional transcriptional effectors downstream of PI3K signaling regulated by a LATS1/2 dependent mechanism. Using in vitro and in vivo approaches, we show that TAZ and YAP are central oncoproteins in PI3K driven oncogenesis along with mTORC1, providing a rationale for combination therapy. Leveraging these findings, we describe a therapeutic approach that builds upon pre-existing therapeutic strategies utilizing mTORC1 inhibitors and combines them with new TEAD inhibitors that target YAP and TAZ. Combination therapy using everolimus and IK-930, an inhibitor targeting autopalmitoylation of the TEADs, synergistically diminished proliferation and anchorage dependent growth of PI3K activated sarcoma cell lines at low, physiologically achievable doses. In vivo, this combination therapy showed a synergistic effect, contrasting with the lack of effect of the individual single agent therapies, suggesting that an integrated view of PI3K and Hippo signaling can be leveraged therapeutically in PI3K activated sarcomas.
2024-10-01 | GSE274982 | GEO
Project description:Tuberous Sclerosis Complex (TSC) is a disease caused by autosomal dominant mutations in the TSC1 or TSC2 genes, and is characterized by tumor susceptibility, brain lesions, seizures and behavioral impairments. The TSC1 and TSC2 genes encode proteins forming a complex (TSC), which is a major regulator and suppressor of mammalian target of rapamycin (mTOR) in complex 1 (mTORC1), a signaling complex that promotes cell growth and proliferation. TSC1/2 loss of heterozygosity (LOH) and the subsequent complete loss of TSC regulatory activity in null cells causes mTORC1 dysregulation and TSC-associated brain lesions or other tissue tumors. However, it is not clear whether TSC1/2 heterozygous brain cells are abnormal and contribute to TSC neuropathology. To investigate this issue, we generated induced pluripotent stem cells (iPSCs) from TSC patients and unaffected controls, and utilized these to obtain neural progenitor cells (NPCs) and differentiated neurons in vitro. These patient-derived TSC2 heterozygous NPCs were delayed in their ability to differentiate into neurons. Patient-derived progenitor cells also exhibited a modest activation of mTORC1 signaling downstream of TSC, and a marked attenuation of upstream PI3K/AKT signaling. We further show that pharmacologic AKT inhibition, but not mTORC1 inhibition, causes a neuronal differentiation delay, mimicking the patient phenotype. Together these data suggest that heterozygous TSC2 mutations disrupt neuronal development, potentially contributing to the disease neuropathology, and that this defect may result from dysregulated AKT signaling in neural progenitor cells.
2018-08-26 | GSE111584 | GEO