Doxorubicin-Induced Elevated Oxidative Stress and Neurochemical Alterations in Brain and Cognitive Decline: Protection by MESNA and Insights into Mechanisms of Chemotherapy-Induced Cognitive Impairment ( Chemobrain )

Chemotherapy-induced cognitive impairment (CICI) is now widely recognized as a real and too common complication of cancer chemotherapy experienced by an ever-growing number of cancer survivors. Previously, we reported that doxorubicin (Dox), a prototypical reactive oxygen species (ROS)-producing anti-cancer drug, results in oxidation of plasma proteins, including apolipoprotein A-I (ApoA-I) leading to tumor necrosis factor-alpha (TNF-α)-mediated oxidative stress in plasma and brain. We also reported that co-administration of the antioxidant drug, 2-mercaptoethane sulfonate sodium (MESNA), prevents Dox-induced protein oxidation and subsequent TNF-α elevation in plasma. In this study, we measured oxidative stress in both brain and plasma of Dox-treated mice both with and without MESNA. MESNA ameliorated Dox-induced oxidative protein damage in plasma, confirming our prior studies, and in a new finding led to decreased oxidative stress in brain. This study also provides further functional and biochemical evidence of the mechanisms of CICI. Using novel object recognition (NOR), we demonstrated the Dox administration resulted in memory deficits, an effect that was rescued by MESNA. Using hydrogen magnetic resonance imaging spectroscopy (H1-MRS) techniques, we demonstrated that Dox administration led to a dramatic decrease in choline-containing compounds assessed by (Cho)/creatine ratios in the hippocampus in mice. To better elucidate a potential mechanism for this MRS observation, we tested the activities of the phospholipase enzymes known to act on phosphatidylcholine (PtdCho), a key component of phospholipid membranes and a source of choline for the neurotransmitter, acetylcholine (ACh). The activities of both phosphatidylcholine-specific phospholipase C (PC-PLC) and phospholipase D were severely diminished following Dox administration. The activity of PC-PLC was preserved when MESNA was co-administered with Dox; however, PLD activity was not protected. This study is the first to demonstrate the protective effects of MESNA on Dox-related protein oxidation, cognitive decline, phosphocholine (PCho) levels, and PC-PLC activity in brain and suggests novel potential therapeutic targets and strategies to mitigate CICI

Metabolic Effects of Acute Thiamine Depletion Are Reversed by Rapamycin in Breast and Leukemia Cells

Thiamine-dependent enzymes (TDEs) control metabolic pathways that are frequently altered in cancer and therefore present cancer-relevant targets. We have previously shown that the recombinant enzyme thiaminase cleaves and depletes intracellular thiamine, has growth inhibitory activity against leukemia and breast cancer cell lines, and that its growth inhibitory effects were reversed in leukemia cell lines by rapamycin. Now, we first show further evidence of thiaminase therapeutic potential by demonstrating its activity against breast and leukemia xenografts, and against a primary leukemia xenograft. We therefore further explored the metabolic effects of thiaminase in combination with rapamycin in leukemia and breast cell lines. Thiaminase decreased oxygen consumption rate and increased extracellular acidification rate, consistent with the inhibitory effect of acute thiamine depletion on the activity of the TDEs pyruvate dehydrogenase and 2-oxoglutarate dehydrogenase complexes; these effects were reversed by rapamycin. Metabolomic studies demonstrated intracellular thiamine depletion and the presence of the thiazole cleavage product in thiaminase-treated cells, providing validation of the experimental procedures. Accumulation of ribose and ribulose in both cell lines support the thiaminase-mediated suppression of the TDE transketolase. Interestingly, thiaminase suppression of another TDE, branched chain amino ketoacid dehydrogenase (BCKDH), showed very different patterns in the two cell lines: in RS4 leukemia cells it led to an increase in BCKDH substrates, and in MCF-7 breast cancer cells it led to a decrease in BCKDH products. Immunoblot analyses showed corresponding differences in expression of BCKDH pathway enzymes, and partial protection of thiaminase growth inhibition by gabapentin indicated that BCKDH inhibition may be a mechanism of thiaminase-mediated toxicity. Surprisingly, most of thiaminase-mediated metabolomic effects were also reversed by rapamycin. Thus, these studies demonstrate that acute intracellular thiamine depletion by recombinant thiaminase results in metabolic changes in thiamine-dependent metabolism, and demonstrate a previously unrecognized role of mTOR signaling in the regulation of thiamine-dependent metabolism

Analysis of test beam data taken with a prototype of TPC with resistive Micromegas for the T2K Near Detector upgrade

In this paper we describe the performance of a prototype of the High Angle Time Projection Chambers (HA-TPCs) that are being produced for the Near Detector (ND280) upgrade of the T2K experiment. The two HA-TPCs of ND280 will be instrumented with eight Encapsulated Resistive Anode Micromegas (ERAM) on each endplate, thus constituting in total 32 ERAMs. This innovative technique allows the detection of the charge emitted by ionization electrons over several pads, improving the determination of the track position. The TPC prototype has been equipped with the first ERAM module produced for T2K and with the HA-TPC readout electronics chain and it has been exposed to the DESY Test Beam in order to measure spatial and dE/dx resolution. In this paper we characterize the performances of the ERAM and, for the first time, we compare them with a newly developed simulation of the detector response. Spatial resolution better than 800 ${\mu \rm m}$ and dE/dx resolution better than 10% are observed for all the incident angles and for all the drift distances of interest. All the main features of the data are correctly reproduced by the simulation and these performances fully fulfill the requirements for the HA-TPCs of T2K

Characterization of Charge Spreading and Gain of Encapsulated Resistive Micromegas Detectors for the Upgrade of the T2K Near Detector Time Projection Chambers

An upgrade of the near detector of the T2K long baseline neutrino oscillation experiment is currently being conducted. This upgrade will include two new Time Projection Chambers, each equipped with 16 charge readout resistive Micromegas modules. A procedure to validate the performance of the detectors at different stages of production has been developed and implemented to ensure a proper and reliable operation of the detectors once installed. A dedicated X-ray test bench is used to characterize the detectors by scanning each pad individually and to precisely measure the uniformity of the gain and the deposited energy resolution over the pad plane. An energy resolution of about 10% is obtained. A detailed physical model has been developed to describe the charge dispersion phenomena in the resistive Micromegas anode. The detailed physical description includes initial ionization, electron drift, diffusion effects and the readout electronics effects. The model provides an excellent characterization of the charge spreading of the experimental measurements and allowed the simultaneous extraction of gain and RC information of the modules

Measurements of the νμ\nu_{\mu} and νˉμ\bar{\nu}_{\mu}-induced Coherent Charged Pion Production Cross Sections on 12C^{12}C by the T2K experiment

We report an updated measurement of the $\nu_{\mu}$-induced, and the first measurement of the $\bar{\nu}_{\mu}$-induced coherent charged pion production cross section on $^{12}C$ nuclei in the T2K experiment. This is measured in a restricted region of the final-state phase space for which $p_{\mu,\pi} > 0.2$ GeV, $\cos(\theta_{\mu}) > 0.8$ and $\cos(\theta_{\pi}) > 0.6$, and at a mean (anti)neutrino energy of 0.85 GeV using the T2K near detector. The measured $\nu_{\mu}$ CC coherent pion production flux-averaged cross section on $^{12}C$ is $(2.98 \pm 0.37 (stat.) \pm 0.31 (syst.) \substack{ +0.49 \\ -0.00 } \mathrm{ (Q^2\,model)}) \times 10^{-40}~\mathrm{cm}^{2}$. The new measurement of the $\bar{\nu}_{\mu}$-induced cross section on $^{12}{C}$ is $(3.05 \pm 0.71 (stat.) \pm 0.39 (syst.) \substack{ +0.74 \\ -0.00 } \mathrm{(Q^2\,model)}) \times 10^{-40}~\mathrm{cm}^{2}$. The results are compatible with both the NEUT 5.4.0 Berger-Sehgal (2009) and GENIE 2.8.0 Rein-Sehgal (2007) model predictions

Measurements of the νμ and ν¯μ -induced coherent charged pion production cross sections on C12 by the T2K experiment

We report an updated measurement of the ν μ -induced, and the first measurement of the ¯ ν μ -induced coherent charged pion production cross section on 12 C nuclei in the Tokai-to-Kamioka experiment. This is measured in a restricted region of the final-state phase space for which p μ , π > 0.2     GeV , cos ( θ μ ) > 0.8 and cos ( θ π ) > 0.6 , and at a mean (anti)neutrino energy of 0.85 GeV using the T2K near detector. The measured ν μ charged current coherent pion production flux-averaged cross section on 12 C is ( 2.98 ± 0.37 ( stat ) ± 0.31 ( syst ) + 0.49 − 0.00 ( Q 2   model ) ) × 10 − 40     cm 2 . The new measurement of the ¯ ν μ -induced cross section on 12 C is ( 3.05 ± 0.71 ( stat ) ± 0.39 ( syst ) + 0.74 − 0.00 ( Q 2   model ) ) × 10 − 40     cm 2 . The results are compatible with both the NEUT 5.4.0 Berger-Sehgal (2009) and GENIE 2.8.0 Rein-Sehgal (2007) model predictions

Basal brain oxidative and nitrative stress levels are finely regulated by the interplay between superoxide dismutase 2 and p53

Superoxide dismutases (SODs) are the primary ROS scavenging enzymes of the cell and catalyze the dismutation of superoxide radicals O2−. to H2O2 and molecular oxygen (O2). Among the three forms of SOD identified manganese-containing SOD (MnSOD, SOD2) is a homotetramer located wholly in the mitochondrial matrix. Due to SOD2 strategic location, it represents the first mechanism of defense against the augmentation of reactive oxygen/reactive nitrogen species (ROS/RNS) levels in the mitochondria in order to prevent further damages. Here, we aimed to understand the effects that the partial lack (SOD2(−/+)) or the overexpression (TgSOD2) of MnSOD produces on oxidative/nitrative stress basal levels in different brain-isolated cellular fractions (i.e. mitochondrial, nuclear, cytosolic) as well as in the whole brain homogenate. Furthermore, due to the known interaction between SOD2 and p53 protein, we aimed to clarify the impact that the double mutation has on oxidative/nitrative stress levels in the brain of the new mice carrying the double mutation (p53(−/−)xSOD2(−/+)) and (p53(−/−)xTgSOD2). Interestingly, we found that each mutation differently affects (i) mitochondrial; (ii) nuclear; and (iii) cytosolic oxidative/nitrative stress basal levels; but, overall, no changes or a reduction of oxidative/nitrative stress levels were found in the whole brain homogenate. Finally, the analysis of well-known antioxidant systems such as thioredoxin-1 and the Nrf2/HO-1/BVR-A system suggested their potential role in the maintenance of the cellular redox homeostasis in the presence of changes of SOD2 and/or p53 protein levels

Differential effects of thiaminase on branched chain amino acid catabolism.

<p>A. Schematic diagram showing branched chain amino acid catabolism. B. Different metabolomic signatures indicating inhibition of BCKDH by thiaminase. The top three panels show accumulation of BCKDH substrates after 48-7 cells after thiaminase treatment- most notably isovalerylcarnitine, which is also reversed by rapamycin. C. An immunoblot of cytosolic and mitochondrial branched chain amino acid transferase (cBCAT and mBCAT), the enzymes that catalyzes the reactions that produce BCKDH substrates, and total and phosphorylated BCKDH subunit E1 (BCKD-E1 and pBCKD-E1, respectively) in RS4 leukemia cells treated with thiaminase, rapamycin or both. For C-48 vs T-48 and C-48 vs T+R-48 comparisons, ** indicates p<0.05 and * indicates 0.05</p