Fatigue in teriflunomide-treated patients with relapsing remitting multiple sclerosis in the real-world Teri-FAST study

BACKGROUND: Fatigue is a frequent and disabling symptom of multiple sclerosis (MS) often associated with impaired quality of life (QoL) in patients. Teriflunomide is a once-daily oral immunomodulator used for the treatment of relapsing remitting forms of MS. However, its effect on fatigue is not well known in real life practice. We evaluated the impact of teriflunomide on fatigue in patients with relapsing remitting MS (RRMS) after 2 years of treatment in the real-world Teri-FAST study. METHODS: Teri-FAST was a 2-year, prospective, observational study conducted in France in RRMS patients treated with teriflunomide 14 mg. Fatigue was assessed using the French version of the modified fatigue impact scale (EMIF-SEP). The primary endpoint was the change from baseline in EMIF-SEP score after 2 years of treatment. Secondary endpoints included evaluation of depression (Beck Depression Inventory [BDI]), health-related QoL (Two-Life Scale TLS-QoL 10), self-reported physical activity, and adverse events. RESULTS: 210 eligible patients were included in the study with a mean age of 45.4 years and a mean ± SD Expanded Disability Status Scale score of 1.76 ± 1.43 at baseline. About half (52.4%) of patients had no previous treatment for MS. In the 163 patients who completed at least 1 follow-up visit, the mean change in EMIF-SEP score at Year 2 was -1.54 (95% CI: -4.02, 0.94) indicating that fatigue remained stable. Similarly, there were no changes in depression level and QoL after 2 years of treatment. Physical activity slightly improved with 57% of patients reporting being physically active after 2 years as compared to 46% at baseline. The safety profile of teriflunomide was consistent with that seen during clinical development, and compliance with treatment was high. CONCLUSION: Fatigue scores remained stable in RRMS patients treated with teriflunomide 14 mg over 2 years in real-life setting. Teriflunomide did not negatively impact depression or QoL

Natalizumab treatment shows low cumulative probabilities of confirmed disability worsening to EDSS milestones in the long-term setting.

Abstract Background Though the Expanded Disability Status Scale (EDSS) is commonly used to assess disability level in relapsing-remitting multiple sclerosis (RRMS), the criteria defining disability progression are used for patients with a wide range of baseline levels of disability in relatively short-term trials. As a result, not all EDSS changes carry the same weight in terms of future disability, and treatment benefits such as decreased risk of reaching particular disability milestones may not be reliably captured. The objectives of this analysis are to assess the probability of confirmed disability worsening to specific EDSS milestones (i.e., EDSS scores ≥3.0, ≥4.0, or ≥6.0) at 288 weeks in the Tysabri Observational Program (TOP) and to examine the impact of relapses occurring during natalizumab therapy in TOP patients who had received natalizumab for ≥24 months. Methods TOP is an ongoing, open-label, observational, prospective study of patients with RRMS in clinical practice. Enrolled patients were naive to natalizumab at treatment initiation or had received ≤3 doses at the time of enrollment. Intravenous natalizumab (300 mg) infusions were given every 4 weeks, and the EDSS was assessed at baseline and every 24 weeks during treatment. Results Of the 4161 patients enrolled in TOP with follow-up of at least 24 months, 3253 patients with available baseline EDSS scores had continued natalizumab treatment and 908 had discontinued (5.4% due to a reported lack of efficacy and 16.4% for other reasons) at the 24-month time point. Those who discontinued due to lack of efficacy had higher baseline EDSS scores (median 4.5 vs. 3.5), higher on-treatment relapse rates (0.82 vs. 0.23), and higher cumulative probabilities of EDSS worsening (16% vs. 9%) at 24 months than those completing therapy. Among 24-month completers, after approximately 5.5 years of natalizumab treatment, the cumulative probabilities of confirmed EDSS worsening by 1.0 and 2.0 points were 18.5% and 7.9%, respectively (24-week confirmation), and 13.5% and 5.3%, respectively (48-week confirmation). The risks of 24- and 48-week confirmed EDSS worsening were significantly higher in patients with on-treatment relapses than in those without relapses. An analysis of time to specific EDSS milestones showed that the probabilities of 48-week confirmed transition from EDSS scores of 0.0–2.0 to ≥3.0, 2.0–3.0 to ≥4.0, and 4.0–5.0 to ≥6.0 at week 288 in TOP were 11.1%, 11.8%, and 9.5%, respectively, with lower probabilities observed among patients without on-treatment relapses (8.1%, 8.4%, and 5.7%, respectively). Conclusions In TOP patients with a median (range) baseline EDSS score of 3.5 (0.0–9.5) who completed 24 months of natalizumab treatment, the rate of 48-week confirmed disability worsening events was below 15%; after approximately 5.5 years of natalizumab treatment, 86.5% and 94.7% of patients did not have EDSS score increases of ≥1.0 or ≥2.0 points, respectively. The presence of relapses was associated with higher rates of overall disability worsening. These results were confirmed by assessing transition to EDSS milestones. Lower rates of overall 48-week confirmed EDSS worsening and of transitioning from EDSS score 4.0–5.0 to ≥6.0 in the absence of relapses suggest that relapses remain a significant driver of disability worsening and that on-treatment relapses in natalizumab-treated patients are of prognostic importance

Finerenone, a Non-Steroidal Mineralocorticoid Receptor Antagonist, Reduces Vascular Injury and Increases Regulatory T-Cells: Studies in Rodents with Diabetic and Neovascular Retinopathy

Vision loss in diabetic retinopathy features damage to the blood–retinal barrier and neovascularization, with hypertension and the renin–angiotensin system (RAS) having causal roles. We evaluated if finerenone, a non-steroidal mineralocorticoid receptor (MR) antagonist, reduced vascular pathology and inflammation in diabetic and neovascular retinopathy. Diabetic and hypertensive transgenic (mRen-2)27 rats overexpressing the RAS received the MR antagonist finerenone (10 mg/kg/day, oral gavage) or the angiotensin-converting enzyme inhibitor perindopril (10 mg/kg/day, drinking water) for 12 weeks. As retinal neovascularization does not develop in diabetic rodents, finerenone (5 mg/kg/day, i.p.) was evaluated in murine oxygen-induced retinopathy (OIR). Retinal vasculopathy was assessed by measuring gliosis, vascular leakage, neovascularization, and VEGF. Inflammation was investigated by quantitating retinal microglia/macrophages, pro-inflammatory mediators, and anti-inflammatory regulatory T-cells (Tregs). In diabetes, both treatments reduced systolic blood pressure, gliosis, vascular leakage, and microglial/macrophage density, but only finerenone lowered VEGF, ICAM-1, and IL-1ß. In OIR, finerenone reduced neovascularization, vascular leakage, and microglial density, and increased Tregs in the blood, spleen, and retina. Our findings, in the context of the FIDELIO-DKD and FIGARO-DKD trials reporting the benefits of finerenone on renal and cardiovascular outcomes in diabetic kidney disease, indicate the potential of finerenone as an effective oral treatment for diabetic retinopathy

Finerenone, a Non-Steroidal Mineralocorticoid Receptor Antagonist, Reduces Vascular Injury and Increases Regulatory T-Cells: Studies in Rodents with Diabetic and Neovascular Retinopathy

Vision loss in diabetic retinopathy features damage to the blood–retinal barrier and neovascularization, with hypertension and the renin–angiotensin system (RAS) having causal roles. We evaluated if finerenone, a non-steroidal mineralocorticoid receptor (MR) antagonist, reduced vascular pathology and inflammation in diabetic and neovascular retinopathy. Diabetic and hypertensive transgenic (mRen-2)27 rats overexpressing the RAS received the MR antagonist finerenone (10 mg/kg/day, oral gavage) or the angiotensin-converting enzyme inhibitor perindopril (10 mg/kg/day, drinking water) for 12 weeks. As retinal neovascularization does not develop in diabetic rodents, finerenone (5 mg/kg/day, i.p.) was evaluated in murine oxygen-induced retinopathy (OIR). Retinal vasculopathy was assessed by measuring gliosis, vascular leakage, neovascularization, and VEGF. Inflammation was investigated by quantitating retinal microglia/macrophages, pro-inflammatory mediators, and anti-inflammatory regulatory T-cells (Tregs). In diabetes, both treatments reduced systolic blood pressure, gliosis, vascular leakage, and microglial/macrophage density, but only finerenone lowered VEGF, ICAM-1, and IL-1ß. In OIR, finerenone reduced neovascularization, vascular leakage, and microglial density, and increased Tregs in the blood, spleen, and retina. Our findings, in the context of the FIDELIO-DKD and FIGARO-DKD trials reporting the benefits of finerenone on renal and cardiovascular outcomes in diabetic kidney disease, indicate the potential of finerenone as an effective oral treatment for diabetic retinopathy

RAP and LRP-1 silencing by siRNA decreased the level and density of lysosome in MCF-7R cells.

MCF-7S, MCF-7R cells that incubated with or without 500 nM RAP for 12 hours were used in this experiment. The endocytic organelles were isolated by density gradient centrifugation as detailed in Materials and Methods. sucrose gradient was analysed using invertase enzyme assay as described in Materials and Methods. Detection of P-gp and Lamp-1 were evaluated by Western-blot in all collected aliquots (A-D). The intensity of the bands was quantified by densitometry using quantity one program. Student’s t-test was used for the statistical significance of different values. ** p < 0.01, *** p < 0.001 for MCF-7R cells versus MCF-7S cells, ## p < 0.01, ### p < 0.001 for MCF-7R treated cells versus MCF-7R untreated cells (E,F,G).</p

Schematic diagram of P-gp internalization by LRP-1 receptor and its implication in MCF-7 cells drug resistance.

Schematic diagram of P-gp internalization by LRP-1 receptor and its implication in MCF-7 cells drug resistance.</p

RAP sensitized MCF-7R cells to Dox cytotoxic effects and reduced IC₅₀.

A and B, MCF-7S and MCF-7R cells were treated with different Dox concentrations (from 0 to 10 μM) with or without 500 nM RAP. After 48 and 72h, cell viability was measured by UptiBlue Viable Cell Counting Assay. The results were presented as percentage of control and represented with standard deviation (S.D.) of at least three independent experiments. Student’s t-test was used for the statistical significance of different values.° NS, *** pC, MCF-7R cells were pretreated with Verapamil (5 μM) for 6h and incubated with Dox (1 μM). After 48 and 72h, cell viability was measured using UptiBlue Viable Cell Assay. The results obtained from three independent experiments (% of control) were represented with standard deviation (S.D.). Student’s t-test was used for the statistical significance of different values. **** pD, summary table of IC50 and RI (resistance Index). IC50 represents the mean half maximal inhibitory concentration. RI was assessed using the quotient of the IC50 values (IC50 MCF-7R/IC50 MCF-7) in each treatment conditions. Student’s t-test was used for the statistical significance of different values. *** p<0.001 for RAP-MCF-7R cells compared to untreated MCF-7R cells.</p

ERK1/2 inhibition reduced Dox cytotoxic effects on MCF-7 cells.

MCF-7S and MCF-7R cells were incubated with 1 μM Dox with or without 500 nM RAP and 1 μM U0126 48 hours. A, Cell viability was measured using UptiBlue Viable Cell Assay. B, Caspase-7 activity was measured by caspACE assay kit. The results obtained from three independent experiments (% of control) were represented with standard deviation (S.D.). Student’s t-test was used for the statistical significance of different values. *** p<0.001 compared to Dox-treated cells and ¥¥¥ p<0.001, ¥¥¥¥ p<0.0001 compared to Dox- and RAP-treated cells.° NS, U0126-treated cells versus Dox-treated cells.</p

LRP-1 silencing by siRNA reduced P-gp expression and sensitized MCF-7R cells to Dox.

A, scRNA-MCF-7R and siRNA-MCF-7R cells were cultured for 24 hours. Detection of P-gp and LRP-1/β chain was evaluated by Western-blot. β-actin antibody was used as a control. The intensity of the bands was quantified by densitometry using quantity one program. Student’s t-test was used for the statistical significance of different values. *** pB, scRNA-MCF-7 and siRNA-MCF-7R cells were incubated with Dox at concentrations ranging from 0 to 10 μM. After 48 and 72h, cell viability was measured using UptiBlue Viable Cell Assay. The results obtained from three independent experiments (% of control) were represented with standard deviation (S.D.). Student’s t-test was used for the statistical significance of different values. *** pC, summary table of IC50. IC50 represents the mean half maximal inhibitory concentration. Student’s t-test was used for the statistical significance of different values. **** p<0.0001 for siRNA-MCF-7R cells compared to scRNA-MCF-7R cells.</p