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Early intervention with a small molecule inhibitor for tumor necrosis factor-α prevents cognitive deficits in a triple transgenic mouse model of Alzheimer's disease.

Gabbita SP, Srivastava MK, Eslami P, Johnson MF, Kobritz NK, Tweedie D, Greig NH, Zemlan FP, Sharma SP, Harris-White ME - J Neuroinflammation (2012)

Bottom Line: Thus, a critical need exists for novel treatments directed towards modifying the pathophysiology and progression.After 10 weeks of treatment, cognitive performance was assessed using radial arm maze and neuroinflammation was assessed using biochemical, stereological and flow cytometric endpoints. 3,6'-dithiothalidomide reduced tumor necrosis factor-α mRNA and protein levels in the brain and improved working memory performance and the ratio of resting to reactive microglia in the hippocampus of triple transgenic mice.These results suggest that modulation of tumor necrosis factor-α with small molecule inhibitors is safe and effective with potential for the long-term prevention and treatment of Alzheimer's disease.

View Article: PubMed Central - HTML - PubMed

Affiliation: P2D Bioscience, Cincinnati, OH 45242, USA.

ABSTRACT

Background: Chronic neuroinflammation is an important component of Alzheimer's disease and could contribute to neuronal dysfunction, injury and loss that lead to disease progression. Multiple clinical studies implicate tumor necrosis factor-α as an inflammatory mediator of neurodegeneration in patients with Alzheimer's because of elevated levels of this cytokine in the cerebrospinal fluid, hippocampus and cortex. Current Alzheimer's disease interventions are symptomatic treatments with limited efficacy that do not address etiology. Thus, a critical need exists for novel treatments directed towards modifying the pathophysiology and progression.

Methods: To investigate the effect of early immune modulation on neuroinflammation and cognitive outcome, we treated triple transgenic Alzheimer's disease mice (harboring PS1(M146V), APP(Swe), and tau(P301L) transgenes) with the small molecule tumor necrosis factor-α inhibitors, 3,6'-dithiothalidomide and thalidomide, beginning at four months of age. At this young age, mice do not exhibit plaque or tau pathology but do show mild intraneuronal amyloid beta protein staining and a robust increase in tumor necrosis factor-α. After 10 weeks of treatment, cognitive performance was assessed using radial arm maze and neuroinflammation was assessed using biochemical, stereological and flow cytometric endpoints.

Results: 3,6'-dithiothalidomide reduced tumor necrosis factor-α mRNA and protein levels in the brain and improved working memory performance and the ratio of resting to reactive microglia in the hippocampus of triple transgenic mice. In comparison to non-transgenic controls, triple transgenic Alzheimer's disease mice had increased total numbers of infiltrating peripheral monomyelocytic/granulocytic leukocytes with enhanced intracytoplasmic tumor necrosis factor-α, which was reduced after treatment with 3,6'-dithiothalidomide.

Conclusions: These results suggest that modulation of tumor necrosis factor-α with small molecule inhibitors is safe and effective with potential for the long-term prevention and treatment of Alzheimer's disease.

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Related in: MedlinePlus

Thalidomide and 3,6′-dithiothalidomide attenuate lipopolysaccharide-induced increase in tumor necrosis factor-α. BV2 cells were treated with 1 ng/mL LPS ± Thal or 3,6′-DT for 24 h. Initial studies in BV2 cells demonstrate that both Thal and 3,6′-DT are effective at attenuating LPS-induced TNFα release into culture media. 3,6′-DT has an IC50 value of approximately 1 μM whereas the IC50 for Thal is > 10 μM. n = 6 per group. One-way analysis of variance: P < 0.0001; *P < 0.001 versus both drugs, both doses. LPS: lipopolysaccharide; Thal: thalidomide; TNFα: tumor necrosis factor-α; 3,6′-DT: 3,6′-dithiothalidomide.
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Figure 2: Thalidomide and 3,6′-dithiothalidomide attenuate lipopolysaccharide-induced increase in tumor necrosis factor-α. BV2 cells were treated with 1 ng/mL LPS ± Thal or 3,6′-DT for 24 h. Initial studies in BV2 cells demonstrate that both Thal and 3,6′-DT are effective at attenuating LPS-induced TNFα release into culture media. 3,6′-DT has an IC50 value of approximately 1 μM whereas the IC50 for Thal is > 10 μM. n = 6 per group. One-way analysis of variance: P < 0.0001; *P < 0.001 versus both drugs, both doses. LPS: lipopolysaccharide; Thal: thalidomide; TNFα: tumor necrosis factor-α; 3,6′-DT: 3,6′-dithiothalidomide.

Mentions: Initial studies were conducted in vitro to verify the efficacy of Thal and 3,6′-DT (see Figure 1 for chemical structures) to inhibit TNFα. BV2 microglial cell cultures were treated with 1 ng/mL LPS with or without Thal or 3,6′-DT (1 and 10 μM). Culture media (n = 6 wells/treatment) was collected 24 h later and evaluated for TNFα protein levels via ELISA (Figure 2) and cytotoxicity by measuring LDH release into the media. One-way ANOVA revealed a significant effect of treatment (F723 = 51.03; P < 0.0001). Both Thal and 3,6′-DT significantly inhibited BV2 TNFα production at both concentrations compared with LPS alone (P < 0.0001 versus both drugs, both doses). 3,6′-DT was a more potent inhibitor, with a half maximal inhibitory concentration (IC50) value for TNFα inhibition of approximately 1 μM while the IC50 value of Thal was in excess of 10 μM, which is congruent with previous publications [36]. There was no increase in LDH in any treatment group including DMSO alone, LPS alone, Thal or 3,6′-DT alone or LPS plus Thal or 3,6′-DT (data not shown).


Early intervention with a small molecule inhibitor for tumor necrosis factor-α prevents cognitive deficits in a triple transgenic mouse model of Alzheimer's disease.

Gabbita SP, Srivastava MK, Eslami P, Johnson MF, Kobritz NK, Tweedie D, Greig NH, Zemlan FP, Sharma SP, Harris-White ME - J Neuroinflammation (2012)

Thalidomide and 3,6′-dithiothalidomide attenuate lipopolysaccharide-induced increase in tumor necrosis factor-α. BV2 cells were treated with 1 ng/mL LPS ± Thal or 3,6′-DT for 24 h. Initial studies in BV2 cells demonstrate that both Thal and 3,6′-DT are effective at attenuating LPS-induced TNFα release into culture media. 3,6′-DT has an IC50 value of approximately 1 μM whereas the IC50 for Thal is > 10 μM. n = 6 per group. One-way analysis of variance: P < 0.0001; *P < 0.001 versus both drugs, both doses. LPS: lipopolysaccharide; Thal: thalidomide; TNFα: tumor necrosis factor-α; 3,6′-DT: 3,6′-dithiothalidomide.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3403851&req=5

Figure 2: Thalidomide and 3,6′-dithiothalidomide attenuate lipopolysaccharide-induced increase in tumor necrosis factor-α. BV2 cells were treated with 1 ng/mL LPS ± Thal or 3,6′-DT for 24 h. Initial studies in BV2 cells demonstrate that both Thal and 3,6′-DT are effective at attenuating LPS-induced TNFα release into culture media. 3,6′-DT has an IC50 value of approximately 1 μM whereas the IC50 for Thal is > 10 μM. n = 6 per group. One-way analysis of variance: P < 0.0001; *P < 0.001 versus both drugs, both doses. LPS: lipopolysaccharide; Thal: thalidomide; TNFα: tumor necrosis factor-α; 3,6′-DT: 3,6′-dithiothalidomide.
Mentions: Initial studies were conducted in vitro to verify the efficacy of Thal and 3,6′-DT (see Figure 1 for chemical structures) to inhibit TNFα. BV2 microglial cell cultures were treated with 1 ng/mL LPS with or without Thal or 3,6′-DT (1 and 10 μM). Culture media (n = 6 wells/treatment) was collected 24 h later and evaluated for TNFα protein levels via ELISA (Figure 2) and cytotoxicity by measuring LDH release into the media. One-way ANOVA revealed a significant effect of treatment (F723 = 51.03; P < 0.0001). Both Thal and 3,6′-DT significantly inhibited BV2 TNFα production at both concentrations compared with LPS alone (P < 0.0001 versus both drugs, both doses). 3,6′-DT was a more potent inhibitor, with a half maximal inhibitory concentration (IC50) value for TNFα inhibition of approximately 1 μM while the IC50 value of Thal was in excess of 10 μM, which is congruent with previous publications [36]. There was no increase in LDH in any treatment group including DMSO alone, LPS alone, Thal or 3,6′-DT alone or LPS plus Thal or 3,6′-DT (data not shown).

Bottom Line: Thus, a critical need exists for novel treatments directed towards modifying the pathophysiology and progression.After 10 weeks of treatment, cognitive performance was assessed using radial arm maze and neuroinflammation was assessed using biochemical, stereological and flow cytometric endpoints. 3,6'-dithiothalidomide reduced tumor necrosis factor-α mRNA and protein levels in the brain and improved working memory performance and the ratio of resting to reactive microglia in the hippocampus of triple transgenic mice.These results suggest that modulation of tumor necrosis factor-α with small molecule inhibitors is safe and effective with potential for the long-term prevention and treatment of Alzheimer's disease.

View Article: PubMed Central - HTML - PubMed

Affiliation: P2D Bioscience, Cincinnati, OH 45242, USA.

ABSTRACT

Background: Chronic neuroinflammation is an important component of Alzheimer's disease and could contribute to neuronal dysfunction, injury and loss that lead to disease progression. Multiple clinical studies implicate tumor necrosis factor-α as an inflammatory mediator of neurodegeneration in patients with Alzheimer's because of elevated levels of this cytokine in the cerebrospinal fluid, hippocampus and cortex. Current Alzheimer's disease interventions are symptomatic treatments with limited efficacy that do not address etiology. Thus, a critical need exists for novel treatments directed towards modifying the pathophysiology and progression.

Methods: To investigate the effect of early immune modulation on neuroinflammation and cognitive outcome, we treated triple transgenic Alzheimer's disease mice (harboring PS1(M146V), APP(Swe), and tau(P301L) transgenes) with the small molecule tumor necrosis factor-α inhibitors, 3,6'-dithiothalidomide and thalidomide, beginning at four months of age. At this young age, mice do not exhibit plaque or tau pathology but do show mild intraneuronal amyloid beta protein staining and a robust increase in tumor necrosis factor-α. After 10 weeks of treatment, cognitive performance was assessed using radial arm maze and neuroinflammation was assessed using biochemical, stereological and flow cytometric endpoints.

Results: 3,6'-dithiothalidomide reduced tumor necrosis factor-α mRNA and protein levels in the brain and improved working memory performance and the ratio of resting to reactive microglia in the hippocampus of triple transgenic mice. In comparison to non-transgenic controls, triple transgenic Alzheimer's disease mice had increased total numbers of infiltrating peripheral monomyelocytic/granulocytic leukocytes with enhanced intracytoplasmic tumor necrosis factor-α, which was reduced after treatment with 3,6'-dithiothalidomide.

Conclusions: These results suggest that modulation of tumor necrosis factor-α with small molecule inhibitors is safe and effective with potential for the long-term prevention and treatment of Alzheimer's disease.

Show MeSH
Related in: MedlinePlus