Dimethyl Fumarate Alleviates Dextran Sulfate Sodium-Induced Colitis, through the Activation of Nrf2-Mediated Antioxidant and Anti-inflammatory Pathways.

Oxidative stress and chronic inflammation play critical roles in the pathogenesis of ulcerative colitis (UC) and inflammatory bowel diseases (IBD). A previous study has demonstrated that dimethyl fumarate (DMF) protects mice from dextran sulfate sodium (DSS)-induced colitis via its potential antioxidant capacity, and by inhibiting the activation of the NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3) inflammasome. This study aims to clarify the nuclear factor erythroid 2-related factor 2/antioxidant responsive element (Nrf2/ARE) pathway pharmacological activation and anti-inflammatory effect by DMF, through focusing on other crucial antioxidant enzymes and inflammatory mediator, including glutamate-cysteine ligase catalytic subunit (GCLC), glutathione peroxidase (GPX) and cyclooxygenase-2 (COX-2), in a DSS-induced colitis mouse model. The oral administration of DMF attenuated the shortening of colons and alleviated colonic inflammation. Furthermore, the expression of key antioxidant enzymes, including GCLC and GPX, in the colonic tissue were significantly increased by DMF administration. In addition, protein expression of the inflammatory mediator, COX-2, was reduced by DMF administration. Our results suggest that DMF alleviates DSS-induced colonic inflammatory damage, likely via up-regulating GCLC and GPX and down-regulating COX-2 protein expression in colonic tissue

Effect of Antioxidants in Cathepsin B Release by HIV Infected Macrophages

During HIV infection of macrophages, the lysosomal protein cathepsin B is released and induces neurotoxicity. Also, the levels of cathepsin B are increased in plasma and post-mortem brain tissue of patients with HIV-associated dementia. Oxidative damage is increased in HIV- infected patients, while antioxidants are decreased in HIV-associated dementia. Dimethyl fumarate (DMF), an antioxidant, has been reported to decrease HIV replication and neurotoxicity caused by HIV-infected macrophages. Since HIV also increases cathepsin B, we hypothesize that DMF will also reduce cathepsin B release from HIV-infected macrophages. Monocyte-derived macrophages (MDM) were isolated from healthy donors and inoculated with HIV-1ADA. After removal of infection, MDM were treated with DMF at different concentrations (15, 30, and 60 µM) until day 12 post-infection, changing and collecting media every three days. HIV-1p24 and cathepsin B levels were assessed from HIV-infected MDM supernatants at the end of cultures using ELISA. Results indicate that DMF reduced HIV-1 replication and cathepsin B secretion from HIV-infected macrophages, in a concentration-dependent manner, in comparison with vehicle (DMSO)-treated controls. However, cathepsin B secretion was not affected by HIV infection in vehicle-treated controls. In conclusion, DMSO may have had an unexpected effect in cathepsin B secretion in our experiments, and this could explain why cathepsin B secretion was not affected by HIV infection. Future experiments will include an untreated control group to determine if DMSO vehicle is having an effect in cathepsin B secretion. This will lead us to determine the role of DMF in cathepsin B secretion from HIV-infected macrophages

Metabolic Inhibitor Effects on Eyespot Formation in Regenerating Planaria

Planaria are flatworms known for their remarkable ability to regenerate. Glycolytic activity has been shown to increase during planarian regeneration (Osuma et al., 2017) and regeneration requires both an increase in mitotic activity and an increase in apoptotic activity to successfully regrow missing structures (Pellettieri et al., 2010; Wenemoser & Reddien, 2010). The objectives of this study are to determine if metabolic inhibitors affect the time needed to regenerate eyespots after head amputation as a measurement of regeneration and if there are any correlations between average numbers of mitotic and apoptotic cells in regenerating planaria in the presence or absence of metabolic inhibitors. Planaria (D. dorotocephala and D. tigrina) were exposed to non-lethal doses of metabolic inhibitors after amputation between the head and the pharynx to determine if metabolic inhibitors affect eyespot regeneration times. Mucus removal, fixation, and bleaching were used to prepare planaria for immunolabeling procedures. Mitotic cells were labeled to determine if metabolic inhibitors affect average numbers of mitotic cells in regenerating planaria. Immunolabelling procedures were used in an attempt to visualize apoptotic cells in planaria. Results from the exposure experiments suggest that the earlier planaria are exposed to metabolic inhibitors after amputation, the longer it takes to regrow eyespots. By comparing the average number of mitotic cells planaria in the presence or absence of metabolic inhibitors, it was determined that delays in eyespot formation are not associated with changes in the average number of mitotic cells. Further studies should be conducted to test for significant differences between apoptotic cells in inhibitor-treated and untreated planaria

Effectiveness of delayed-release dimethyl fumarate on patient-reported outcomes and clinical measures in patients with relapsing-remitting multiple sclerosis in a real-world clinical setting: PROTEC.

Ensaio clínico PROTEC, Protocolo nº 109MS408Abstract BACKGROUND: Patient-reported outcomes (PRO) and clinical outcomes give a broad assessment of relapsing-remitting multiple sclerosis (RRMS) disease. OBJECTIVE: The aim is to evaluate the effectiveness of delayed-release dimethyl fumarate (DMF) on disease activity and PROs in patients with RRMS in the clinic. METHODS: PROTEC, a phase 4, open-label, 12-month observational study, assessed annualized relapse rate (ARR), proportion of patients relapsed, and changes in PROs. Newly diagnosed and early MS (≤3.5 EDSS and ≤1 relapse in the prior year) patient subgroups were evaluated. RESULTS: Unadjusted ARR at 12 months post-DMF versus 12 months before DMF initiation was 75% lower (0.161 vs. 0.643, p < 0.0001) overall (n = 1105) and 84%, 77%, and 71% lower in newly diagnosed, ≤3.5 EDSS, and ≤1 relapse subgroups, respectively. Overall, 88% of patients were relapse-free 12 months after DMF initiation (84%, newly diagnosed; 88%, ≤3.5 EDSS; 88%, ≤1 relapse). PRO measures for fatigue, treatment satisfaction, daily living, and work improved significantly over 12 months of DMF versus baseline. CONCLUSION: At 12 months after versus 12 months before DMF initiation, ARR was significantly lower, the majority of patients were relapse-free, and multiple PRO measures showed improvement (overall and for subgroups), suggesting that DMF is effective based on clinical outcomes and from a patient perspective.Clinical trial: A Study Evaluating the Effectiveness of Tecfidera (Dimethyl Fumarate) on Multiple Sclerosis (MS) Disease Activity and Patient-Reported Outcomes (PROTEC), NCT01930708,info:eu-repo/semantics/publishedVersio

Effect of DMF on liver I/R

AIM To investigate the hypothesis that treatment with dimethyl fumarate (DMF) may ameliorate liver ischemia/reperfusion injury (I/RI). METHODS Rats were divided into 3 groups: sham, control (CTL), and DMF. DMF (25 mg/kg, twice/d) was orally administered for 2 d before the procedure. The CTL and DMF rats were subjected to ischemia for 1 h and reperfusion for 2 h. The serum alanine aminotransferase (ALT) and malondialdehyde (MDA) levels, adenosine triphosphate (ATP), NO × metabolites, anti-oxidant enzyme expression level, anti-inflammatory effect, and anti-apoptotic effect were determined. RESULTS Histological tissue damage was significantly reduced in the DMF group (Suzuki scores: sham: 0 ± 0; CTL: 9.3 ± 0.5; DMF: 2.5 ± 1.2; sham vs CTL, P < 0.0001; CTL vs DMF, P < 0.0001). This effect was associated with significantly lower serum ALT (DMF 5026 ± 2305 U/L vs CTL 10592 ± 1152 U/L, P = 0.04) and MDA (DMF 18.2 ± 1.4 μmol/L vs CTL 26.0 ± 1.0 μmol/L, P = 0.0009). DMF effectively improved the ATP content (DMF 20.3 ± 0.4 nmol/mg vs CTL 18.3 ± 0.6 nmol/mg, P = 0.02), myeloperoxidase activity (DMF 7.8 ± 0.4 mU/mL vs CTL 6.0 ± 0.5 mU/mL, P = 0.01) and level of endothelial nitric oxide synthase expression (DMF 0.38 ± 0.05-fold vs 0.17 ± 0.06-fold, P = 0.02). The higher expression levels of anti-oxidant enzymes (catalase and glutamate-cysteine ligase modifier subunit and lower levels of key inflammatory mediators (nuclear factor-kappa B and cyclooxygenase-2 were confirmed in the DMF group. CONCLUSION DMF improved the liver function and the anti-oxidant and inflammation status following I/RI. Treatment with DMF could be a promising strategy in patients with liver I/RI

Effects of dimethyl fumarate on neuroprotection and immunomodulation

BACKGROUND: Neuronal degeneration in multiple sclerosis has been linked to oxidative stress. Dimethyl fumarate is a promising novel oral therapeutic option shown to reduce disease activity and progression in patients with relapsing-remitting multiple sclerosis. These effects are presumed to originate from a combination of immunomodulatory and neuroprotective mechanisms. We aimed to clarify whether neuroprotective concentrations of dimethyl fumarate have immunomodulatory effects. FINDINGS: We determined time- and concentration-dependent effects of dimethyl fumarate and its metabolite monomethyl fumarate on viability in a model of endogenous neuronal oxidative stress and clarified the mechanism of action by quantitating cellular glutathione content and recycling, nuclear translocation of transcription factors, and the expression of antioxidant genes. We compared this with changes in the cytokine profiles released by stimulated splenocytes measured by ELISPOT technology and analyzed the interactions between neuronal and immune cells and neuronal function and viability in cell death assays and multi-electrode arrays. Our observations show that dimethyl fumarate causes short-lived oxidative stress, which leads to increased levels and nuclear localization of the transcription factor nuclear factor erythroid 2-related factor 2 and a subsequent increase in glutathione synthesis and recycling in neuronal cells. Concentrations that were cytoprotective in neuronal cells had no negative effects on viability of splenocytes but suppressed the production of proinflammatory cytokines in cultures from C57BL/6 and SJL mice and had no effects on neuronal activity in multi-electrode arrays. CONCLUSIONS: These results suggest that immunomodulatory concentrations of dimethyl fumarate can reduce oxidative stress without altering neuronal network activity

An Investigation of the Succination of the Selenoproteins Thioredoxin Reductase and Glutathione Peroxidase

We have identified the irreversible protein modification S-(2-succino)cysteine, a product of the Kreb’s cycle intermediate fumarate reacting with the thiol group of cysteine (Cys) residues, also termed succination. Succination is increased in adipose tissue of diabetic mice in vivo and in adipocytes matured in high glucose medium (25 mM) in vitro. Selenocysteine (Sec) is a less common but highly conserved amino acid that has major functions in redox chemistry. Sec possesses a lower pKa (~5.4) and is much more reactive in comparison to Cys. This project aimed to determine if succination occurred on Sec residues in two selenoproteins that play roles of maintaining intracellular redox environments and contain Sec in their active sites, thioredoxin reductase (TrxR) and glutathione peroxidase (GPx). We hypothesized that increased fumarate levels will lead to the succination of Sec and causing decreased enzymatic activity for each enzyme. Two cellular models were used with elevated fumarate levels: (1) 3T3-L1 adipocytes matured in high glucose medium (25 mM) to mimic diabetic conditions and (2) 3T3-L1 fibroblasts where fumarase was knocked-down (Fh k/d) to mimic fumarase deficient cancers. As a positive control to demonstrate the ability to succinate TrxR in vitro, recombinant TrxR1 was incubated with the reactive fumarate ester, dimethyl fumarate (DMF). After analysis, a ~56% reduction in activity was detected due to succination of an active site residue, which was confirmed by mass spectrometric analyses. Unexpectedly, TrxR activity was found to be significantly increased in 3T3-L1 adipocytes matured in 25 mM glucose compared to 5 mM glucose, despite no changes in protein levels. No statistically significant changes in TrxR activity were found in Fh k/d 3T3-L1 fibroblasts compared to controls. GPx activity was found to be significantly reduced in 3T3-L1 adipocytes matured in high glucose with no changes in levels of GPx protein. GPx activity was also analyzed in Fh k/d 3T3-L1 fibroblasts, however no significant changes were detected. Overall, the results demonstrate that TrxR can be modified readily by reactive fumarate esters, leading to decreased enzymatic activity. However, endogenously produced fumarate does not appear to significantly contribute to TrxR or GPx modification in the models studied

Dimethyl fumarate: a new oral treatment option for multiple sclerosis

Multiple Sclerosis (MS) is a slowly progressive, immunologically mediated disease of the CNS. The recent years have witnessed great efforts in establishing new therapeutic options for multiple sclerosis. There is a clear need for more effective, safe and at the same time orally available treatment options. Here we review the recently approved drug Dimethyl fumarate (DMF, Tecfidera®) as a new therapeutic option for MS and its role in context to the existing oral treatment options for MS. Dimethyl fumarate is the methyl ester of fumaric acid and has been claimed to possess immunomodulatory properties and is already in clinical use as Fumaderm for severe systemic psoriasis. In addition, Dimethyl fumarate was also shown to act on the blood-brain barrier and exert neuroprotective properties via activation of anti-oxidative pathways and displayed beneficial effects in experimental autoimmune encephalomyelitis (EAE), a model mimicking many aspects of MS. Based on two global phase III studies. Dimethyl fumarate has been clinically proven to significantly reduce important measures of disease activity, including relapses and development of brain lesions, as well as to slow disability progression over time, while demonstrating a favourable safety and tolerability profile

Dimethyl fumarate in the management of multiple sclerosis: Appropriate patient selection and special considerations

Delayed-release dimethyl fumarate (DMF), also known as gastroresistant DMF, is the most recently approved oral disease-modifying treatment (DMT) for relapsing multiple sclerosis. Two randomized clinical trials (Determination of the Efficacy and Safety of Oral Fumarate in Relapsing–Remitting MS [DEFINE] and Comparator and an Oral Fumarate in Relapsing-Remitting Multiple Sclerosis [CONFIRM]) demonstrated significant efficacy in reducing relapse rate and radiological signs of disease activity, as seen on magnetic resonance imaging. The DEFINE study also indicated a significant effect of DMF on disability worsening, while the low incidence of confirmed disability worsening in the CONFIRM trial rendered an insignificant reduction among the DMF-treated groups when compared to placebo. DMF also demonstrated a good safety profile and acceptable tolerability, since the most common side effects (gastrointestinal events and flushing reactions) are usually transient and mild to moderate in severity. Here, we discuss the place in therapy of DMF for individuals with relapsing multiple sclerosis, providing a tentative therapeutic algorithm to manage newly diagnosed patients and those who do not adequately respond to self-injectable DMTs. Literature data supporting the potential role of DMF as a first-line therapy are presented. The possibility of using DMF as switching treatment or even as an add-on strategy in patients with breakthrough disease despite self-injectable DMTs will also be discussed. Lastly, we argue about the role of DMF as an exit strategy from natalizumab-treated patients who are considered at risk for developing multifocal progressive leukoencephalopathy