Performance Analysis of IRS-Assisted THz Communication Systems over α-µ Fading Channels with Pointing Errors.

In this paper, we analyze the performance of an intelligent reflecting surface (IRS)-aided terahertz (THz) wireless communication system with pointing errors. Specifically, we derive closed-form analytical expressions for the upper bounded ergodic capacity and approximate expression of the outage probability. We adopt an α-µ fading channel model for our analysis that is experimentally demonstrated to be a good fit for THz small-scale fading statistics, especially in indoor communication scenarios. In the proposed analysis, the statistical distribution of the α-µ fading channel is used to derive analytical expressions for the ergodic capacity and outage probability. Our proposed analysis considers not only the IRS reflected channels, but also the direct channel between the communication nodes. The results of the derived analytical expressions are validated through Monte Carlo simulations. Through simulations, it has been noticed that pointing errors degrade the performance of the IRS-assisted THz wireless communication system which can be compensated by deploying an IRS having a large number of reflecting elements.

Mechanistic Target of Rapamycin (mTOR) Inhibition Synergizes with Reduced Internal Ribosome Entry Site (IRES)-mediated Translation of Cyclin D1 and c-MYC mRNAs to Treat Glioblastoma.

Our previous work has demonstrated an intrinsic mRNA-specific protein synthesis salvage pathway operative in glioblastoma (GBM) tumor cells that is resistant to mechanistic target of rapamycin (mTOR) inhibitors. The activation of this internal ribosome entry site (IRES)-dependent mRNA translation initiation pathway results in continued translation of critical transcripts involved in cell cycle progression in the face of global eIF-4E-mediated translation inhibition. Recently we identified compound 11 (C11), a small molecule capable of inhibiting c-MYC IRES translation as a consequence of blocking the interaction of a requisite c-MYC IRES trans-acting factor, heterogeneous nuclear ribonucleoprotein A1, with its IRES. Here we demonstrate that C11 also blocks cyclin D1 IRES-dependent initiation and demonstrates synergistic anti-GBM properties when combined with the mechanistic target of rapamycin kinase inhibitor PP242. The structure-activity relationship of C11 was investigated and resulted in the identification of IRES-J007, which displayed improved IRES-dependent initiation blockade and synergistic anti-GBM effects with PP242. Mechanistic studies with C11 and IRES-J007 revealed binding of the inhibitors within the UP1 fragment of heterogeneous nuclear ribonucleoprotein A1, and docking analysis suggested a small pocket within close proximity to RRM2 as the potential binding site. We further demonstrate that co-therapy with IRES-J007 and PP242 significantly reduces tumor growth of GBM xenografts in mice and that combined inhibitor treatments markedly reduce the mRNA translational state of cyclin D1 and c-MYC transcripts in these tumors. These data support the combined use of IRES-J007 and PP242 to achieve synergistic antitumor responses in GBM.

Phosphorylation of the Hippo Pathway Component AMOTL2 by the mTORC2 Kinase Promotes YAP Signaling, Resulting in Enhanced Glioblastoma Growth and Invasiveness.

The mechanistic target of rapamycin (mTOR) and Hippo signaling pathways are two major signaling cascades that coordinately regulate cell growth and proliferation. Dysregulation of these pathways plays a critical role in gliomagenesis. Recent reports have provided evidence of cross-talk between the mTOR and Hippo pathways; however, a complete description of the signaling relationships between these pathways remains to be elucidated. Utilizing a gene-trapping strategy in a mouse glioma model, we report the identification of AMOTL2 as a candidate substrate for mTORC2. AMOTL2 is phosphorylated at serine 760 by mTORC2. Mutation of AMOTL2 mimicking constitutive Ser(760) phosphorylation blocks its ability to bind and repress YAP leading to increased relative expression of known YAP gene targets. Moreover, overexpression of AMOTL2 or a nonphosphorylatable AMOTL2-S760A mutant inhibited YAP-induced transcription, foci formation, growth, and metastatic properties, whereas overexpression of a phosphomimetic AMOTL2-S760E mutant negated these repressive effects of AMOTL2 in glioblastoma (GBM) cells in vitro. Similar effects on xenograft growth were observed in GBM cells expressing these AMOTL2 Ser(760) mutants. YAP was also shown to be required for Rictor-mediated GBM growth and survival. Finally, an analysis of mTORC2/AMOTL2/YAP activities in primary GBM samples supported the clinical relevance of this signaling cascade, and we propose that pharmacological agents cotargeting these regulatory circuits may hold therapeutic potential.

Engineered MXene/Bi2S3 nanoflowers in sodium alginate hydrogel: A synergistic eradicator of disinfected byproducts in aqueous environment.

In this work, Bi2S3 nanoflowers were in situ anchored on the surface of Ti3C2 via a hydrothermal process to obtain MXene-supported Ti3C2/Bi2S3 nanocomposite, then incorporated inside in sodium alginate polymer to prepared hydrogel materials (Ti3C2/Bi2S3@SA-H) which outperforms and have an excellent capability for the removal of pollutants like disinfected byproducts. The synthesized hydrogel material Ti3C2/Bi2S3@SA-H may be utilized for a variety of functional materials in environmental applications. Furthermore, the Ti3C2/Bi2S3@SA-H was characterized by SEM, EDX, XRD, BET, AFM, FTIR, Zeta potential, XPS, Raman and TGA. Remarkably, Ti3C2/Bi2S3@SA-H hydrogel 0.007 cm3 g-1, 159.5 nm and 0.0017 cm3 g-1, 160.5 nm materials exhibited the highest average pore diameter. The research focused on evaluating the adsorption capability of Ti3C2/Bi2S3@SA-H hydrogel materials for 2,6-dibromo-4-nitrophenol (DBNP), 2,4,6-triiodophenol (TIP), 2,4,6-Trichlorophenol (TCP) and 2,6-dichloro-4-nitrophenol (DCNP). The findings indicated that the material exhibited the eradication efficiency of about 662, 657, 647 and 617 mg/g from DBNP, TIP, TCP and DCNP respectively. Several adsorption isotherms were extensively examined, encompassing the Temkin, Langmuir and Freundlich models, alongside pseudo-first and second-order models. The Langmuir and pseudo-second-order models showed the highest degree of consistency with the observed data. Concerning regeneration and reusability, the materials demonstrated easy regeneration and effective recyclability over the course of 10 cycles. The notable adsorption capacity, coupled with the innovative combination of Ti3C2/Bi2S3 and polymer hydrogel, along with its recyclability, positions our material Ti3C2/Bi2S3@SA-H as a highly prospective competitors for wastewater treatment and other critical areas in water research.

Dry gel spinning of fungal hydrogels for the development of renewable yarns from food waste.

BACKGROUND: Renewable materials made using environmentally friendly processes are in high demand as a solution to reduce the pollution created by the fashion industry. In recent years, there has been a growing trend in research on renewable materials focused on bio-based materials derived from fungi. RESULTS: Recently, fungal cell wall material of a chitosan producing fungus has been wet spun to monofilaments. This paper presents a modification for the fungal monofilament spinning process, by the development of a benign method, dry gel spinning, to produce continuous monofilaments and twisted multifilament yarns, from fungal cell wall, that can be used in textile applications. The fungal biomass of Rhizopus delemar, grown using bread waste as a substrate, was subjected to alkali treatment with a dilute sodium hydroxide solution to isolate alkali-insoluble material (AIM), which mainly consists of the fungal cell wall. The treatment of AIM with dilute lactic acid resulted in hydrogel formation. The morphology of the hydrogels was pH dependent, and they exhibited shear thinning viscoelastic behavior. Dry gel spinning of the fungal hydrogels was first conducted using a simple lab-scale syringe pump to inject the hydrogels through a needle to form a monofilament, which was directly placed on a rotating receiver and left to dry at room temperature. The resulting monofilament was used to make twisted multifilament yarns. The process was then improved by incorporating a heated chamber for the quicker drying of the monofilaments (at 30°C). Finally, the spinning process was scaled up using a twin-screw microcompounder instead of the syringe pump. The monofilaments were several meters long and reached a tensile strength of 63 MPa with a % elongation at break of 14. When spinning was performed in the heated chamber, the tensile strength increased to 80 MPa and further increased to 103 MPa when a micro-compounder was used for spinning. CONCLUSION: The developed dry gel spinning method shows promising results in scalability and demonstrates the potential for renewable material production using fungi. This novel approach produces materials with mechanical properties comparable to those of conventional textile fibers.

Fabrication of maleic anhydride-acrylamide copolymer based sodium alginate hydrogel for elimination of metals ions and dyes contaminants from polluted water.

The study explores the synergy of biobased polymers and hydrogels for water purification. Polymer nanomaterial's, synthesized by combining acrylamide copolymer with maleic anhydride, were integrated into sodium alginate biopolymer using an eco-friendly approach. Crosslinking agents, calcium chloride and glutaraladehyde, facilitated seamless integration, ensuring non-toxicity, high adsorption performance, and controlled capacity. This innovative combination presents a promising solution for clean and healthy water supplies, addressing the critical need for sustainable environmental practices in water purification. In addition, the polymer sodium alginate hydrogel (MAH@AA-P/SA/H) underwent characterization via the use of several analytical procedures, such as FTIR, XPS, SEM, EDX and XRD. Adsorption studies were conducted on metals and dyes in water, and pollutant removal methods were explored. We investigated several variables (such as pH, starting concentration, duration, and absorbent quantity) affect a material's capacity to be adsorbed. Moreover, the maximum adsorption towards Cu2+ is 754 mg/g while for Cr6+ metal ions are 738 mg/g, while the adsorption towards Congo Red and Methylene Blue dye are 685 mg/g and 653 mg/g correspondingly, within 240 min. Adsorption results were further analyzed using kinetic and isothermal models, which showed that MAH@AA-P/SA/H adsorption is governed by a chemisorption process. Hence, the polymer prepared from sodium alginate hydrogel (MAH@AA-P/SA/H) has remarkable properties as a versatile material for the significantly elimination of harmful contaminants from dirty water.

A Ni-doped Mo2C/NCF composite for efficient electrocatalytic hydrogen evolution.

Ni-Mo2C nano catalysts dispersed on N-doped carbon flowers: a composite with nitrogen-containing carbon flowers carrying nickel-modified molybdenum carbide exhibits enhanced HER catalytic activity in alkaline electrolyte.

Phosphomimetic substitution of heterogeneous nuclear ribonucleoprotein A1 at serine 199 abolishes AKT-dependent internal ribosome entry site-transacting factor (ITAF) function via effects on strand annealing and results in mammalian target of rapamycin complex 1 (mTORC1) inhibitor sensitivity.

The relative activity of the AKT kinase has been demonstrated to be a major determinant of sensitivity of tumor cells to mammalian target of rapamycin (mTOR) complex 1 inhibitors. Our previous studies have shown that the multifunctional RNA-binding protein heterogeneous nuclear ribonucleoprotein (hnRNP) A1 regulates a salvage pathway facilitating internal ribosome entry site (IRES)-dependent mRNA translation of critical cellular determinants in an AKT-dependent manner following mTOR inhibitor exposure. This pathway functions by stimulating IRES-dependent translation in cells with relatively quiescent AKT, resulting in resistance to rapamycin. However, the pathway is repressed in cells with elevated AKT activity, rendering them sensitive to rapamycin-induced G(1) arrest as a result of the inhibition of global eIF-4E-mediated translation. AKT phosphorylation of hnRNP A1 at serine 199 has been demonstrated to inhibit IRES-mediated translation initiation. Here we describe a phosphomimetic mutant of hnRNP A1 (S199E) that is capable of binding both the cyclin D1 and c-MYC IRES RNAs in vitro but lacks nucleic acid annealing activity, resulting in inhibition of IRES function in dicistronic mRNA reporter assays. Utilizing cells in which AKT is conditionally active, we demonstrate that overexpression of this mutant renders quiescent AKT-containing cells sensitive to rapamycin in vitro and in xenografts. We also demonstrate that activated AKT is strongly correlated with elevated Ser(P)(199)-hnRNP A1 levels in a panel of 22 glioblastomas. These data demonstrate that the phosphorylation status of hnRNP A1 serine 199 regulates the AKT-dependent sensitivity of cells to rapamycin and functionally links IRES-transacting factor annealing activity to cellular responses to mTOR complex 1 inhibition.

Electro-Synthesis of Organic Compounds with Heterogeneous Catalysis.

Electro-organic synthesis has attracted a lot of attention in pharmaceutical science, medicinal chemistry, and future industrial applications in energy storage and conversion. To date, there has not been a detailed review on electro-organic synthesis with the strategy of heterogeneous catalysis. In this review, the most recent advances in synthesizing value-added chemicals by heterogeneous catalysis are summarized. An overview of electrocatalytic oxidation and reduction processes as well as paired electrocatalysis is provided, and the anodic oxidation of alcohols (monohydric and polyhydric), aldehydes, and amines are discussed. This review also provides in-depth insight into the cathodic reduction of carboxylates, carbon dioxide, CC, C≡C, and reductive coupling reactions. Moreover, the electrocatalytic paired electro-synthesis methods, including parallel paired, sequential divergent paired, and convergent paired electrolysis, are summarized. Additionally, the strategies developed to achieve high electrosynthesis efficiency and the associated challenges are also addressed. It is believed that electro-organic synthesis is a promising direction of organic electrochemistry, offering numerous opportunities to develop new organic reaction methods.

Hippocampal adult neurogenesis is enhanced by chronic eszopiclone treatment in rats.

The adult hippocampal dentate gyrus (DG) exhibits cell proliferation and neurogenesis throughout life. We examined the effects of daily administration of eszopiclone (Esz), a commonly used hypnotic drug and gamma-aminobutyric acid (GABA) agonist, compared with vehicle, on DG cell proliferation and neurogenesis, and on sleep-wake patterns. Esz was administered during the usual sleep period of rats, to mimic typical use in humans. Esz treatment for 7 days did not affect the rate of cell proliferation, as measured by 5-bromo-2'-deoxyuridine (BrdU) immunostaining. However, twice-daily Esz administration for 2 weeks increased survival of newborn cells by 46%. Most surviving cells exhibited a neuronal phenotype, identified as BrdU-neuronal nuclei (NeuN) double-labeling. NeuN is a marker of neurons. Non-rapid eye movement sleep was increased on day 1, but not on days 7 or 14 of Esz administration. Delta electroencephalogram activity was increased on days 1 and 7 of treatment, but not on day 14. There is evidence that enhancement of DG neurogenesis is a critical component of the effects of antidepressant treatments of major depressive disorder (MDD). Adult-born DG cells are responsive to GABAergic stimulation, which promotes cell maturation. The present study suggests that Esz, presumably acting as a GABA agonist, has pro-neurogenic effects in the adult DG. This result is consistent with evidence that Esz enhances the antidepressant treatment response of patients with MDD with insomnia.

Melt rheology and extrudate swell properties of talc filled polyethylene compounds.

An experimental study of high-density polyethylene (HDPE) composites filled with talc (0-15 wt.%) was carried out to investigate the rheological properties. The apparent melt viscosity, melt density, and die-swell ratio (B) of the composites were measured at constant shear stress and constant shear rate by using a melt flow indexer and capillary rheometer. The experimental conditions were set to a temperature range from 190 to 220 °C for both apparatuses whereas a load range from 5 to 12.16 kg was selected for melt flow indexer and shear rate range from 1 to 10000 s-1 for capillary rheometer. The initial study showed that the talc particulates did not influence the melt viscosity compared with the neat HDPE but decreased the elasticity of the polymer system. The HDPE/talc systems obeyed power-law model in shear stress-shear rate variations and were shear thinning, meanwhile, the die-swell increased with an increased wall shear rate and shear stress. The melt density of the composites increased linearly with an increase of the filler weight fraction and decreased with the increase of the testing temperature. The talc-HDPE composites showed compressible in the molten state.

Degraded image enhancement by image dehazing and Directional Filter Banks using Depth Image based Rendering for future free-view 3D-TV.

DIBR-3D technology has evolved over the past few years with the demands of consumers increasing in recent times for future free-view 3D videos on their home televisions. The main issue in 3D technology is the lack of 3D content available to watch using the traditional TV systems. Although, some sophisticated devices like stereoscopic cameras have been used to fill the gap between the 3D content demand and 3D content supply. But the content generated through these sophisticated devices can not be displayed on the traditional TV systems, so there needs to be some mechanism which is inline with the traditional TV. Furthermore, the huge collection of existing 2D content should be converted to 3D using depth image-based rendering techniques. This conversion technique can highly contribute in overcoming the shortage problem of the 3D content. This paper presents a novel approach for converting 2D degraded image for DIBR 3D-TV view. This degraded or noisy/blur image is enhanced through image dehazing and Directional Filter Bank (DFB). This enhancement is necessary because of the occlusion effect or hole filling problem that occurs due to imperfect depth map. The enhanced image is then segmented into the foreground image and the background image. After the segmentation, the depth map is generated using image profiles. Moreover, Stereoscopic images are finally produced using the DIBR procedure which is based on the 2D input image and the corresponding depth map. We have verified the results of the proposed approach by comparing the results with the existing state-of-the-art techniques.

Correction: Specific blockade of Rictor-mTOR association inhibits mTORC2 activity and is cytotoxic in glioblastoma.

[This corrects the article DOI: 10.1371/journal.pone.0176599.].

Rapid eye movement sleep deprivation contributes to reduction of neurogenesis in the hippocampal dentate gyrus of the adult rat.

STUDY OBJECTIVES: The dentate gyrus (DG) of the adult hippocampus contains progenitor cells, which have potential to differentiate into neurons. Previously we reported that 96 hours of total sleep deprivation reduces neurogenesis in the DG of adult rats. Loss of either non-rapid eye movement (NREM) or rapid eye movement (REM) sleep could have contributed to the effect of total sleep deprivation. The present study assessed the effect of 4 days of REM sleep deprivation (REMD) on neurogenesis. DESIGN: REMD was achieved by brief treadmill movement initiated by automatic online detection of REM sleep. A yoked-control (YC) rat was placed in the same treadmill and experienced the identical movement regardless the stage of the sleep-wake cycle. The thymidine analog 5- bromo- 2'- deoxy-uridine and the intrinsic proliferation marker, Ki-67, were both used to label proliferating cells. SETTING: Basic neurophysiology laboratory. PARTICIPANTS: Male Sprague-Dawley male rats (300-320 g). RESULTS: REM sleep was reduced by 85% in REMD rats and by 43% in YC, compared with cage control animals and by 79% in REMD rats compared with YC. NREM sleep and slow wave activity within NREM did not differ in REMD and YC groups. Cell proliferation was reduced by 63 % in REMD compared with YC rats, and by 82% and 51%, respectively, in REMD and YC rats compared with cage controls. Across all animals, cell proliferation exhibited a positive correlation with the percentage of REM sleep (r = 0.84, P < 0.001). Reduced cell proliferation in REMD rats was confirmed with the intrinsic proliferation marker, Ki-67. REMD also reduced the percentage of proliferating cells that later expressed a mature neuronal marker. CONCLUSIONS: The present findings support a hypothesis that REM sleep-associated processes facilitate proliferation of granule cells in the adult hippocampal DG.

Inactivation of median preoptic nucleus causes c-Fos expression in hypocretin- and serotonin-containing neurons in anesthetized rat.

The median preoptic nucleus (MnPN) of the hypothalamus contains sleep-active neurons including sleep-active GABAergic neurons and is involved in the regulation of nonREM/REM sleep. The hypocretinergic (HCRT) neurons of the perifornical-lateral hypothalamic area (PF-LHA) and serotonergic (5-HT) neurons of the dorsal raphe nucleus (DRN) are mostly active during waking and have been implicated in the regulation of arousal. MnPN GABAergic neurons project to the PF-LHA and DRN. It is hypothesized that MnPN promotes sleep by inhibiting multiple arousal systems including HCRT and other wake-active neurons within the PF-LHA and 5-HT neurons in the DRN. We examined the effects of inactivation of MnPN neurons by locally microinjecting 0.2 microl of 1 mM or 10 mM solutions of a GABA(A) receptor agonist, muscimol, into the MnPN on Fos expression (Fos-IR) in the PF-LHA neurons including HCRT neurons and 5-HT neurons in the DRN in anesthetized rats. Compared to artificial cerebrospinal fluid control, microinjection of muscimol into the MnPN resulted in significantly higher percentages of HCRT and non-HCRT neurons in the PF-LHA and 5-HT neurons in the DRN that exhibited Fos-IR. The percentage of melanin-concentrating hormone (MCH)+/Fos+ neurons in the PF-LHA did not change after muscimol treatments. These results support a hypothesis that the activation of MnPN neurons contributes to the suppression of wake-promoting systems including HCRT and other unidentified neurons in the PF-LHA and 5-HT neurons in the DRN. These results also suggest that MCH neurons may not be under MnPN inhibitory control. These findings are consistent with a hypothesized role of MnPN in sleep regulation.

Cell proliferation in the dentate gyrus of the adult rat fluctuates with the light-dark cycle.

This study measured cell proliferation in the hippocampal dentate gyrus in the adult rat at different times within a 12:12h light-dark cycle. The experiments were conducted in animals living in either a complex environment or in standard lab cages. A single dose of the thymidine analog 5-Bromo-2'-deoxyuridine (BrdU) was injected 2h before animals were sacrificed either 4, 11, 16, or 23h after the beginning of the light phase of the light-dark cycle (designated ZT0). In both studies, we found a significant increase in the number of BrdU-positive cells in the subgranular cell layer (SGZ) following BrdU administration at ZT9 and sacrifice at ZT11, compared to other circadian times examined. BrdU administration at ZT9 was timed to primarily identify proliferating cells that were in the S phase of the cell cycle during the light phase. Our results suggest that cell proliferation is enhanced either by sleep or by other variables coupled to the light phase of the circadian cycle.

Specific blockade of Rictor-mTOR association inhibits mTORC2 activity and is cytotoxic in glioblastoma.

A small molecule which specifically blocks the interaction of Rictor and mTOR was identified utilizing a high-throughput yeast two-hybrid screen and evaluated as a potential inhibitor of mTORC2 activity in glioblastoma multiforme (GBM). In vitro, CID613034 inhibited mTORC2 kinase activity at submicromolar concentrations and in cellular assays specifically inhibited phosphorylation of mTORC2 substrates, including AKT (Ser-473), NDRG1 (Thr-346) and PKCα (Ser-657), while having no appreciable effects on the phosphorylation status of the mTORC1 substrate S6K (Thr-389) or mTORC1-dependent negative feedback loops. CID613034 demonstrated significant inhibitory effects on cell growth, motility and invasiveness in GBM cell lines and sensitivity correlated with relative Rictor or SIN1 expression. Structure-activity relationship analyses afforded an inhibitor, JR-AB2-011, with improved anti-GBM properties and blocked mTORC2 signaling and Rictor association with mTOR at lower effective concentrations. In GBM xenograft studies, JR-AB2-011 demonstrated significant anti-tumor properties. These data support mTORC2 as a viable therapeutic target in GBM and suggest that targeting protein-protein interactions critical for mTORC2 function is an effective strategy to achieve therapeutic responses.

mTORC2 modulates feedback regulation of p38 MAPK activity via DUSP10/MKP5 to confer differential responses to PP242 in glioblastoma.

Dual-specificity phosphatases (DUSPs) dephosphorylate MAP kinases (MAPKs) resulting in their inactivation. Activation of MAPK signaling leads to enhanced DUSP expression, thus establishing feedback regulation of the MAPK pathway. The DUSPs are subject to regulation at the post-translational level via phosphorylation resulting in alterations of protein stability. Here we report that mTORC2 function leads to stabilization of the p38 MAPK phosphatase, DUSP10, thereby inhibiting p38 activity. We demonstrate that mTORC2 binds DUSP10 and phosphorylates DUSP10 on serine residues 224 and 230. These phosphorylation events block DUSP10 turnover resulting in inactivation of p38 signaling. We further show that insulin-stimulated PI3K/mTORC2 signaling regulates DUSP10 stability and p38 activity. Importantly, knockdown of DUSP10 or ectopic overexpression of nonphosphorylatable or phosphomimetic DUSP10 mutants was sufficient to confer differential mTOR kinase inhibitor responses to GBM cells in vitro and in murine xenografts. Finally, DUSP10 was shown to be overexpressed in a significant number of GBM patients. These data demonstrate the ability of the mTORC2 pathway to exert regulatory effects on the DUSP10/p38 feedback loop to control the cellular effects of mTOR kinase inhibitors in GBM and support the use of DUSP10 expression as a surrogate biomarker to predict responsiveness.

Protor-2 interacts with tristetraprolin to regulate mRNA stability during stress.

The A/U-rich RNA-binding protein tristetraprolin (TTP) is an mRNA destabilizing factor which plays a role in the regulated turnover of many transcripts encoding proteins involved in immune function and cell growth control. TTP also plays a role in stress-induced destabilization of mRNAs. Here we report the interaction of TTP with a component of the mTORC2 kinase, Protor-2 (PRR5-L, protein Q6MZQ0/FLJ14213/CAE45978). Protor-2 is structurally similar to human PRR5 and has been demonstrated to bind mTORC2 via Rictor and/or Sin1 and may signal downstream events promoting apoptosis. Protor-2 dissociates from mTORC2 upon hyperactivation of the kinase and is not required for mTORC2 integrity or activity. We identified Protor-2 in a yeast two-hybrid screen as a TTP interactor using the C-terminal mRNA decay domain of TTP as bait. The interaction of Protor-2 with TTP was also confirmed in vivo in co-immunoprecipitation experiments and Protor-2 was also detected in immunoprecipitates of Rictor. Protor-2 was shown to stimulate TTP-mediated mRNA turnover of several TTP-associated mRNAs (TNF-α, GM-CSF, IL-3 and COX-2) in Jurkat cells when overexpressed while the half-lives of transcripts which do not decay via a TTP-mediated mechanism were unaffected. Knockdown of Protor-2 via RNAi inhibited TTP-mediated mRNA turnover of these TTP-associated mRNAs and inhibited association of TTP with cytoplasmic stress granules (SG) or mRNA processing bodies (P-bodies) following induction of the integrated stress response. These results suggest that Protor-2 associates with TTP to accelerate TTP-mediated mRNA turnover and functionally links the control of TTP-regulated mRNA stability to mTORC2 activity.

Conditional astroglial Rictor overexpression induces malignant glioma in mice.

BACKGROUND: Hyperactivation of the mTORC2 signaling pathway has been shown to contribute to the oncogenic properties of gliomas. Moreover, overexpression of the mTORC2 regulatory subunit Rictor has been associated with increased proliferation and invasive character of these tumor cells. METHODOLOGY/PRINCIPAL FINDINGS: To determine whether Rictor overexpression was sufficient to induce glioma formation in mice, we inserted a Cre-lox-regulated human Rictor transgene into the murine ROSA26 locus. This floxed Rictor strain was crossed with mice expressing the Cre recombinase driven from the glial fibrillary acidic protein (GFAP) promoter whose expression is limited to the glial cell compartment. Double transgenic GFAP-Cre/Rictor(loxP/loxP) mice developed multifocal infiltrating glioma containing elevated mTORC2 activity and typically involved the subventricular zone (SVZ) and lateral ventricle. Analysis of Rictor-dependent signaling in these tumors demonstrated that in addition to elevated mTORC2 activity, an mTORC2-independent marker of cortical actin network function, was also elevated. Upon histological examination of the neoplasms, many displayed oligodendroglioma-like phenotypes and expressed markers associated with oligodendroglial lineage tumors. To determine whether upstream oncogenic EGFRvIII signaling would alter tumor phenotypes observed in the GFAP-Cre/Rictor(loxP/loxP) mice, transgenic GFAP-EGFRvIII; GFAP-Cre/Rictor(loxP/loxP) mice were generated. These mice developed mixed astrocytic-oligodendroglial tumors, however glioma formation was accelerated and correlated with increased mTORC2 activity. Additionally, the subventricular zone within the GFAP-Cre/Rictor(loxP/loxP) mouse brain was markedly expanded, and a further proliferation within this compartment of the brain was observed in transgenic GFAP-EGFRvIII; GFAP-Cre/Rictor(loxP/loxP) mice. CONCLUSION/SIGNIFICANCE: These data collectively establish Rictor as a novel oncoprotein and support the role of dysregulated Rictor expression in gliomagenesis via mTOR-dependent and mTOR-independent mechanisms. Furthermore, oncogenic EGFRvIII signaling appears to potentiate the in vivo proliferative capacity of GFAP-Cre/Rictor(loxP/loxP) gliomas.