Targeting PI3K/Akt/mTOR in AML: Rationale and Clinical Evidence.

Acute myeloid leukemia (AML) is a highly heterogeneous hematopoietic malignancy characterized by excessive proliferation and accumulation of immature myeloid blasts in the bone marrow. AML has a very poor 5-year survival rate of just 16% in the UK; hence, more efficacious, tolerable, and targeted therapy is required. Persistent leukemia stem cell (LSC) populations underlie patient relapse and development of resistance to therapy. Identification of critical oncogenic signaling pathways in AML LSC may provide new avenues for novel therapeutic strategies. The phosphatidylinositol-3-kinase (PI3K)/Akt and the mammalian target of rapamycin (mTOR) signaling pathway, is often hyperactivated in AML, required to sustain the oncogenic potential of LSCs. Growing evidence suggests that targeting key components of this pathway may represent an effective treatment to kill AML LSCs. Despite this, accruing significant body of scientific knowledge, PI3K/Akt/mTOR inhibitors have not translated into clinical practice. In this article, we review the laboratory-based evidence of the critical role of PI3K/Akt/mTOR pathway in AML, and outcomes from current clinical studies using PI3K/Akt/mTOR inhibitors. Based on these results, we discuss the putative mechanisms of resistance to PI3K/Akt/mTOR inhibition, offering rationale for potential candidate combination therapies incorporating PI3K/Akt/mTOR inhibitors for precision medicine in AML

Multiparametric assessment of mitochondrial respiratory inhibition in HepG2 and RPTEC/TERT1 cells using a panel of mitochondrial targeting agrochemicals

Evidence is mounting for the central role of mitochondrial dysfunction in several pathologies including metabolic diseases, accelerated ageing, neurodegenerative diseases and in certain xenobiotic-induced organ toxicity. Assessing mitochondrial perturbations is not trivial and the outcomes of such investigations are dependent on the cell types used and assays employed. Here we systematically investigated the effect of electron transport chain (ETC) inhibitors on multiple mitochondrial-related parameters in two human cell types, HepG2 and RPTEC/TERT1. Cells were exposed to a broad range of concentrations of 20 ETC-inhibiting agrochemicals and capsaicin, consisting of inhibitors of NADH dehydrogenase (Complex I, CI), succinate dehydrogenase (Complex II, CII) and cytochrome bc1 complex (Complex III, CIII). A battery of tests was utilised, including viability assays, lactate production, mitochondrial membrane potential (MMP) and the Seahorse bioanalyser, which simultaneously measures extracellular acidification rate [ECAR] and oxygen consumption rate [OCR]. CI inhibitors caused a potent decrease in OCR, decreased mitochondrial membrane potential, increased ECAR and increased lactate production in both cell types. Twenty-fourhour exposure to CI inhibitors decreased viability of RPTEC/TERT1 cells and 3D spheroid-cultured HepG2 cells in the presence of glucose. CI inhibitors decreased 2D HepG2 viability only in the absence of glucose. CII inhibitors had no notable effects in intact cells up to 10 µM. CIII inhibitors had similar effects to the CI inhibitors. Antimycin A was the most potent CIII inhibitor, with activity in the nanomolar range. The proposed CIII inhibitor cyazofamid demonstrated a mitochondrial uncoupling signal in both cell types. The study presents a comprehensive example of a mitochondrial assessment workflow and establishes measurable key events of ETC inhibition.Toxicolog

Mapping the cellular response to electron transport chain inhibitors reveals selective signaling networks triggered by mitochondrial perturbation

Mitochondrial perturbation is a key event in chemical-induced organ toxicities that is incompletely understood. Here, we studied how electron transport chain (ETC) complex I, II, or III (CI, CII and CIII) inhibitors affect mitochondrial functionality, stress response activation, and cell viability using a combination of high-content imaging and TempO-Seq in HepG2 hepatocyte cells. CI and CIII inhibitors perturbed mitochondrial membrane potential (MMP) and mitochondrial and cellular ATP levels in a concentration- and time-dependent fashion and, under conditions preventing a switch to glycolysis attenuated cell viability, whereas CII inhibitors had no effect. TempO-Seq analysis of changes in mRNA expression pointed to a shared cellular response to CI and CIII inhibition. First, to define specific ETC inhibition responses, a gene set responsive toward ETC inhibition (and not to genotoxic, oxidative, or endoplasmic reticulum stress) was identified using targeted TempO-Seq in HepG2. Silencing of one of these genes, NOS3, exacerbated the impact of CI and CIII inhibitors on cell viability, indicating its functional implication in cellular responses to mitochondrial stress. Then by monitoring dynamic responses to ETC inhibition using a HepG2 GFP reporter panel for different classes of stress response pathways and applying pathway and gene network analysis to TempO-Seq data, we looked for downstream cellular events of ETC inhibition and identified the amino acid response (AAR) as being triggered in HepG2 by ETC inhibition. Through in silico approaches we provide evidence indicating that a similar AAR is associated with exposure to mitochondrial toxicants in primary human hepatocytes. Altogether, we (i) unravel quantitative, time- and concentration-resolved cellular responses to mitochondrial perturbation, (ii) identify a gene set associated with adaptation to exposure to active ETC inhibitors, and (iii) show that ER stress and an AAR accompany ETC inhibition in HepG2 and primary hepatocytes.Toxicolog

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Energetics of the dipole flip-flop motion in a ferroelectric polymer chain

The authors report on a study of dipole flip-flop “local” transition in ferroelectric polyvinylidene fluoride [P(VDF)] chains, using total energy calculation based on the density functional theory. The calculated results indicate that a simple flipping of a single electric dipole moment is energetically allowed. Furthermore, such a flipping involves no change either in bond length, bond angle, or the orientation of the chain. The calculations also show that on a thin film of ordered chains, strong dipole interactions existing in P(VDF) could cause modulation of the dipole orientation thus forming superlattices on P(VDF) films. These results are in good agreement with recent scanning tunnel microscope experimental measurements. Furthermore, our calculations show that partial flipping may also exist and extend over a length of several monomers during the flip-flop transition

How titanium dioxide and zinc oxide nanoparticles do affect soil microorganism activity?

The increasing use of nanoparticles (NPs) in recent years has posed a potential threat to the natural ecosystem. The aim of this study was to determine the C and N mineralizations of different textured (clay and sandy) soils at increasing doses of (25, 50 and 100 mg kg -1 ) TiO 2 NPs and ZnO NPs that are biological synthesized from Peganum harmala L. plant extract, and to reveal their potential effects on the soil ecosystem. The carbon (C) and nitrogen (N) mineralizations were respectively determined by the CO 2 respiration method and the Parnas-Wagner method under the controlled laboratory conditions for 42 days (28 °C, 80% of field capacity). The highest carbon mineralization among all applications was in clay soils mixed with 100 mg kg -1 TiO 2 NPs. In clay soils treated with ZnO NPs, the treatment of 50 mg kg -1 ZnO NPs increased microorganism activity (P < 0.001). Carbon mineralization rates were found to be higher in sandy soils compared to clay soils (P < 0.05). It was observed that these rates were higher in the TiO 2 NPs treated soils compared to the ZnO NPs treated soils. The nitrogen mineralization ratios were found to be higher in clay soils compared to sandy soils. All of these results indicate that the microorganisms affected from both the presence of TiO 2 NPs and ZnO NPs and differences of texture in these two soils. © 2019 Elsevier Masson SA

Soil organic matter mineralization under different temperatures and moisture conditions in Kizildag plateau, Turkey

Drought by climate change in East Mediterranean Region will change soil temperature and moisture that lead to alter the cycling of biological elements like carbon and nitrogen. However, there are few studies that show how sensitivity of soil organic matter mineralization to temperature and/or moisture can be modified by changes in these parameters. In order to study how these changes in temperature and moisture affect soil carbon and nitrogen mineralization, a laboratory experiment was carried out in two depths (0-5 and 5-15 cm) of soils of Onobrychis beata and Trifolium speciosum being common annual plants in Turkey that was taken from Kizildag Plataeu (Adana city). Some soil physical and chemical properties and as well as rate of carbon and nitrogen mineralizations were determined for both depths of soils. These soils were incubated for 42 days under different field capacities (FC 60, 80 and 100%) and temperatures (24, 28 and 32°C). Cumulative carbon mineralization (Cm), potential mineralizable carbon (C0) and rate of carbon mineralization of all soils were increased with rising temperatures. Rate of carbon mineralization in O. beata soil were lower than T. speciosum soil. NH4-N and NO3-N contents at 42nd day were higher than initial levels of soils and also increased with temperatures and field capacities. In summary, sensitivity of soil organic matter mineralization to temperature was higher at 32°C in upper layer and lower at 24°C in deeper layer of both soils