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Early-stage treatment with Withaferin A reduces levels of misfolded superoxide dismutase 1 and extends lifespan in a mouse model of amyotrophic lateral sclerosis.

Patel P, Julien JP, Kriz J - Neurotherapeutics (2015)

Bottom Line: Approximately 20% of cases of familial amyotrophic lateral sclerosis (ALS) are caused by mutations in the gene encoding Cu/Zn superoxide dismutase (SOD1).The beneficial effects of WA in the SOD1(G93A) mice model were accompanied by an alleviation of neuroinflammation, a decrease in levels of misfolded SOD1 species in the spinal cord, and a reduction in loss of motor neurons resulting in delayed disease progression and mortality.These results suggest that WA may represent a potential lead compound for drug development aiming to treat ALS.

View Article: PubMed Central - PubMed

Affiliation: Research Centre of Institut Universitaire en Santé Mentale de Québec, and Department of Psychiatry and Neuroscience, Laval University, 2601 Chemin de la Canardière, Québec, QC, G1J 2G3, Canada.

ABSTRACT
Approximately 20% of cases of familial amyotrophic lateral sclerosis (ALS) are caused by mutations in the gene encoding Cu/Zn superoxide dismutase (SOD1). Recent studies have shown that Withaferin A (WA), an inhibitor of nuclear factor-kappa B activity, was efficient in reducing disease phenotype in a TAR DNA binding protein 43 transgenic mouse model of ALS. These findings led us to test WA in mice from 2 transgenic lines expressing different ALS-linked SOD1 mutations, SOD1(G93A) and SOD1(G37R). Intraperitoneal administration of WA at a dosage of 4 mg/kg of body weight was initiated from postnatal day 40 until end stage in SOD1(G93A) mice, and from 9 months until end stage in SOD1(G37R) mice. The beneficial effects of WA in the SOD1(G93A) mice model were accompanied by an alleviation of neuroinflammation, a decrease in levels of misfolded SOD1 species in the spinal cord, and a reduction in loss of motor neurons resulting in delayed disease progression and mortality. Interestingly, WA treatment triggered robust induction of heat shock protein 25 (a mouse ortholog of heat shock protein 27), which may explain the reduced level of misfolded SOD1 species in the spinal cord of SOD1(G93A) mice and the decrease of neuronal injury responses, as revealed by real-time imaging of biophotonic SOD1(G93A) mice expressing a luciferase transgene under the control of the growth-associated protein 43 promoter. These results suggest that WA may represent a potential lead compound for drug development aiming to treat ALS.

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No change in proliferation and polarization of peripheral immune cells in superoxide dismutase 1 (SOD1)G93A mice after Withaferin A treatment. Topographic representation of CD4+CD25+Foxp3+ regulatory T (Treg) cells over CD45+ population of (A)WA treated mice and (B) the control group, as analyzed by flow cytometry. (C) Flow cytometry analysis showed no difference in Treg population between the treated and control group at 112 or 125 days. Topographic representation of mean (D, E) interleukin (IL)-10 and (G, H) IL-4 intensity in Tregs of WA-treated and control SOD1G93A mice. Analysis of (F) IL-10 and (I) IL-4 mean intensity in Treg cells in the WA-treated and control groups showed no difference at any mentioned time point. (J, K) No change in the CD4+ and CD8+ T lymphocyte population at any given time point. (L) Fluorescence-activated cell sorting analysis showed a tendency for a higher CD11b+ population in the WA-treated group at 112 days (WA: 15.80 ± 1.30; vehicle: 10.69 ± 1.80; p =0.08) but not at 125 days (WA: 10.42 ± 2.10; vehicle: 9.20 ± 0.50; p =0.73). FITC = fluorescein isothiocyanate; PE = phycoerythrin
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Fig6: No change in proliferation and polarization of peripheral immune cells in superoxide dismutase 1 (SOD1)G93A mice after Withaferin A treatment. Topographic representation of CD4+CD25+Foxp3+ regulatory T (Treg) cells over CD45+ population of (A)WA treated mice and (B) the control group, as analyzed by flow cytometry. (C) Flow cytometry analysis showed no difference in Treg population between the treated and control group at 112 or 125 days. Topographic representation of mean (D, E) interleukin (IL)-10 and (G, H) IL-4 intensity in Tregs of WA-treated and control SOD1G93A mice. Analysis of (F) IL-10 and (I) IL-4 mean intensity in Treg cells in the WA-treated and control groups showed no difference at any mentioned time point. (J, K) No change in the CD4+ and CD8+ T lymphocyte population at any given time point. (L) Fluorescence-activated cell sorting analysis showed a tendency for a higher CD11b+ population in the WA-treated group at 112 days (WA: 15.80 ± 1.30; vehicle: 10.69 ± 1.80; p =0.08) but not at 125 days (WA: 10.42 ± 2.10; vehicle: 9.20 ± 0.50; p =0.73). FITC = fluorescein isothiocyanate; PE = phycoerythrin

Mentions: Evidence suggests that WA may affect the ratio and polarization properties of peripheral myeloid cells, including macrophages and T-cells. One of the particular concerns was the potential effects of WA on the subpopulation of regulatory T cells (Tregs) [61]. Namely, it has been well documented that there is an alteration in the population of T lymphocytes (specifically Tregs) in the blood of patients with ALS and in ALS mouse models [62–66]. Tregs are critically involved in suppressing inflammation induced by neurotoxic T lymphocytes and microglia/macrophages, and they play a prominent role in slowing the rate of disease progression in ALS mice [67–70]. Therefore, we analyzed the number of Tregs in the blood by fluorescence-activated cell sorting (FACS) in the WA-treated and nontreated SOD1G93A mice at two time points, postnatal day 112 and postnatal day 125. As Tregs can express anti-inflammatory cytokines, we also measured the levels of IL-4 and IL-10. The Treg transcription factor FoxP3 is currently the most reliable marker for identifying Tregs. Therefore, CD4+ CD25+FoxP3+ Tregs from the WA-treated and control SOD1G93A mice were quantified. As shown in Fig. 6 (A–C) there was no significant difference in the number of Treg cells from the groups of animals at 112 or 125 days. Likewise, there was no difference in the levels of IL-10 (Fig. 6D–F) and IL-4 (Fig. 6G–I) between WA-treated and control group. In addition, we also investigated for possible shift in CD4+ and CD8+, as well as the CD11b+, population. The quantitative FACS analysis revealed no changes in the CD4+ and CD8+ T lymphocyte population at any mentioned time point (Fig. 6J,K). At the initial time point of 112 days we observed a slight and transient increase in the CD11b+ population. However, the observed changes did not reach statistical significance (Fig. 6L).Fig. 6


Early-stage treatment with Withaferin A reduces levels of misfolded superoxide dismutase 1 and extends lifespan in a mouse model of amyotrophic lateral sclerosis.

Patel P, Julien JP, Kriz J - Neurotherapeutics (2015)

No change in proliferation and polarization of peripheral immune cells in superoxide dismutase 1 (SOD1)G93A mice after Withaferin A treatment. Topographic representation of CD4+CD25+Foxp3+ regulatory T (Treg) cells over CD45+ population of (A)WA treated mice and (B) the control group, as analyzed by flow cytometry. (C) Flow cytometry analysis showed no difference in Treg population between the treated and control group at 112 or 125 days. Topographic representation of mean (D, E) interleukin (IL)-10 and (G, H) IL-4 intensity in Tregs of WA-treated and control SOD1G93A mice. Analysis of (F) IL-10 and (I) IL-4 mean intensity in Treg cells in the WA-treated and control groups showed no difference at any mentioned time point. (J, K) No change in the CD4+ and CD8+ T lymphocyte population at any given time point. (L) Fluorescence-activated cell sorting analysis showed a tendency for a higher CD11b+ population in the WA-treated group at 112 days (WA: 15.80 ± 1.30; vehicle: 10.69 ± 1.80; p =0.08) but not at 125 days (WA: 10.42 ± 2.10; vehicle: 9.20 ± 0.50; p =0.73). FITC = fluorescein isothiocyanate; PE = phycoerythrin
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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Fig6: No change in proliferation and polarization of peripheral immune cells in superoxide dismutase 1 (SOD1)G93A mice after Withaferin A treatment. Topographic representation of CD4+CD25+Foxp3+ regulatory T (Treg) cells over CD45+ population of (A)WA treated mice and (B) the control group, as analyzed by flow cytometry. (C) Flow cytometry analysis showed no difference in Treg population between the treated and control group at 112 or 125 days. Topographic representation of mean (D, E) interleukin (IL)-10 and (G, H) IL-4 intensity in Tregs of WA-treated and control SOD1G93A mice. Analysis of (F) IL-10 and (I) IL-4 mean intensity in Treg cells in the WA-treated and control groups showed no difference at any mentioned time point. (J, K) No change in the CD4+ and CD8+ T lymphocyte population at any given time point. (L) Fluorescence-activated cell sorting analysis showed a tendency for a higher CD11b+ population in the WA-treated group at 112 days (WA: 15.80 ± 1.30; vehicle: 10.69 ± 1.80; p =0.08) but not at 125 days (WA: 10.42 ± 2.10; vehicle: 9.20 ± 0.50; p =0.73). FITC = fluorescein isothiocyanate; PE = phycoerythrin
Mentions: Evidence suggests that WA may affect the ratio and polarization properties of peripheral myeloid cells, including macrophages and T-cells. One of the particular concerns was the potential effects of WA on the subpopulation of regulatory T cells (Tregs) [61]. Namely, it has been well documented that there is an alteration in the population of T lymphocytes (specifically Tregs) in the blood of patients with ALS and in ALS mouse models [62–66]. Tregs are critically involved in suppressing inflammation induced by neurotoxic T lymphocytes and microglia/macrophages, and they play a prominent role in slowing the rate of disease progression in ALS mice [67–70]. Therefore, we analyzed the number of Tregs in the blood by fluorescence-activated cell sorting (FACS) in the WA-treated and nontreated SOD1G93A mice at two time points, postnatal day 112 and postnatal day 125. As Tregs can express anti-inflammatory cytokines, we also measured the levels of IL-4 and IL-10. The Treg transcription factor FoxP3 is currently the most reliable marker for identifying Tregs. Therefore, CD4+ CD25+FoxP3+ Tregs from the WA-treated and control SOD1G93A mice were quantified. As shown in Fig. 6 (A–C) there was no significant difference in the number of Treg cells from the groups of animals at 112 or 125 days. Likewise, there was no difference in the levels of IL-10 (Fig. 6D–F) and IL-4 (Fig. 6G–I) between WA-treated and control group. In addition, we also investigated for possible shift in CD4+ and CD8+, as well as the CD11b+, population. The quantitative FACS analysis revealed no changes in the CD4+ and CD8+ T lymphocyte population at any mentioned time point (Fig. 6J,K). At the initial time point of 112 days we observed a slight and transient increase in the CD11b+ population. However, the observed changes did not reach statistical significance (Fig. 6L).Fig. 6

Bottom Line: Approximately 20% of cases of familial amyotrophic lateral sclerosis (ALS) are caused by mutations in the gene encoding Cu/Zn superoxide dismutase (SOD1).The beneficial effects of WA in the SOD1(G93A) mice model were accompanied by an alleviation of neuroinflammation, a decrease in levels of misfolded SOD1 species in the spinal cord, and a reduction in loss of motor neurons resulting in delayed disease progression and mortality.These results suggest that WA may represent a potential lead compound for drug development aiming to treat ALS.

View Article: PubMed Central - PubMed

Affiliation: Research Centre of Institut Universitaire en Santé Mentale de Québec, and Department of Psychiatry and Neuroscience, Laval University, 2601 Chemin de la Canardière, Québec, QC, G1J 2G3, Canada.

ABSTRACT
Approximately 20% of cases of familial amyotrophic lateral sclerosis (ALS) are caused by mutations in the gene encoding Cu/Zn superoxide dismutase (SOD1). Recent studies have shown that Withaferin A (WA), an inhibitor of nuclear factor-kappa B activity, was efficient in reducing disease phenotype in a TAR DNA binding protein 43 transgenic mouse model of ALS. These findings led us to test WA in mice from 2 transgenic lines expressing different ALS-linked SOD1 mutations, SOD1(G93A) and SOD1(G37R). Intraperitoneal administration of WA at a dosage of 4 mg/kg of body weight was initiated from postnatal day 40 until end stage in SOD1(G93A) mice, and from 9 months until end stage in SOD1(G37R) mice. The beneficial effects of WA in the SOD1(G93A) mice model were accompanied by an alleviation of neuroinflammation, a decrease in levels of misfolded SOD1 species in the spinal cord, and a reduction in loss of motor neurons resulting in delayed disease progression and mortality. Interestingly, WA treatment triggered robust induction of heat shock protein 25 (a mouse ortholog of heat shock protein 27), which may explain the reduced level of misfolded SOD1 species in the spinal cord of SOD1(G93A) mice and the decrease of neuronal injury responses, as revealed by real-time imaging of biophotonic SOD1(G93A) mice expressing a luciferase transgene under the control of the growth-associated protein 43 promoter. These results suggest that WA may represent a potential lead compound for drug development aiming to treat ALS.

Show MeSH
Related in: MedlinePlus