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The tyrphostin agent AG490 prevents and reverses type 1 diabetes in NOD mice.

Davoodi-Semiromi A, Wasserfall CH, Xia CQ, Cooper-DeHoff RM, Wabitsch M, Clare-Salzler M, Atkinson M - PLoS ONE (2012)

Bottom Line: However, the therapeutic effects of AG490 on the development of T1D are unknown.AG490 significantly inhibited the development of T1D (p = 0.02, p = 0.005; at two different time points).The use of such agents, given their extensive safety profiles, provides a strong foundation for their translation to humans with or at increased risk for the disease.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacotherapy & Translational Research, College of Pharmacy, University of Florida, Gainesville, Florida, United States of America. dsemiromi@cop.ufl.edu

ABSTRACT

Background: Recent studies in the NOD (non-obese diabetic) mouse model of type 1 diabetes (T1D) support the notion that tyrosine kinase inhibitors have the potential for modulating disease development. However, the therapeutic effects of AG490 on the development of T1D are unknown.

Materials and methods: Female NOD mice were treated with AG490 (i.p, 1 mg/mouse) or DMSO starting at either 4 or 8 week of age, for five consecutive week, then once per week for 5 additional week. Analyses for the development and/or reversal of diabetes, insulitis, adoptive transfer, and other mechanistic studies were performed.

Results: AG490 significantly inhibited the development of T1D (p = 0.02, p = 0.005; at two different time points). Monotherapy of newly diagnosed diabetic NOD mice with AG490 markedly resulted in disease remission in treated animals (n = 23) in comparision to the absolute inability (0%; 0/10, p = 0.003, Log-rank test) of DMSO and sustained eugluycemia was maintained for several months following drug withdrawal. Interestingly, adoptive transfer of splenocytes from AG490 treated NOD mice failed to transfer diabetes to recipient NOD.Scid mice. CD4 T-cells as well as bone marrow derived dendritic cells (BMDCs) from AG490 treated mice, showed higher expression of Foxp3 (p<0.004) and lower expression of co-stimulatory molecules, respectively. Screening of the mouse immune response gene arrary indicates that expression of costimulaotry molecule Ctla4 was upregulated in CD4+ T-cell in NOD mice treated with AG490, suggesting that AG490 is not a negative regulator of the immune system.

Conclusion: The use of such agents, given their extensive safety profiles, provides a strong foundation for their translation to humans with or at increased risk for the disease.

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Related in: MedlinePlus

In vivo AG490 treated splenocytes did not transfer diabetes to NOD.Scid mice.Prediabetic NOD mice (4- weeks of age) were treated with AG490 or DMSO for five consecutive week (n = 5 per group), three times per week and mice were sacrificed one week after the last injection at week 10. Splenocyte cell suspension of pooled mice immediately prepared and dead cells and red blood cells were removed by Histopacque gradient separation method and freshly isolated splenocytes containing >99% viable cells were transferred (5×106, i.p) into NOD.Scid mice. Blood glucose in recipient mice was monitored at least once a week as described earlier (*: p = 0.02, Kaplan-Meier test).
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pone-0036079-g003: In vivo AG490 treated splenocytes did not transfer diabetes to NOD.Scid mice.Prediabetic NOD mice (4- weeks of age) were treated with AG490 or DMSO for five consecutive week (n = 5 per group), three times per week and mice were sacrificed one week after the last injection at week 10. Splenocyte cell suspension of pooled mice immediately prepared and dead cells and red blood cells were removed by Histopacque gradient separation method and freshly isolated splenocytes containing >99% viable cells were transferred (5×106, i.p) into NOD.Scid mice. Blood glucose in recipient mice was monitored at least once a week as described earlier (*: p = 0.02, Kaplan-Meier test).

Mentions: In order to understand how AG490 reversed and blocked the onset of autoimmune diabetes in vivo, we treated prediabetic NOD mice (4-week of age) with AG490 or with DMSO for 5 week. Mice were sacrificied at week 10 and splenocyte cell suspensions were prepared with Histopaque gradient separation. The dead cells and reticulocytes were removed by the Histopaque gradient separation, therefore, dead cells were excluded without magnetic purification and without activation of T-cells and cell viability was greater than 99%. Five million freshely prepared of pooled splenocytes of NOD mice treated with AG490 (n = 5) or DMSO (n = 5) were transferred by i.p injection into recipient NOD.Scid mice and blood glucose and body weight was closely monitored. Our data as shown in figure 3 suggests that prevention of autoimmune diabetes by AG490 could be due to inactivation of atuoreactive T-cells and/or induction/generation of regulatory T-cell or other regulatory population.


The tyrphostin agent AG490 prevents and reverses type 1 diabetes in NOD mice.

Davoodi-Semiromi A, Wasserfall CH, Xia CQ, Cooper-DeHoff RM, Wabitsch M, Clare-Salzler M, Atkinson M - PLoS ONE (2012)

In vivo AG490 treated splenocytes did not transfer diabetes to NOD.Scid mice.Prediabetic NOD mice (4- weeks of age) were treated with AG490 or DMSO for five consecutive week (n = 5 per group), three times per week and mice were sacrificed one week after the last injection at week 10. Splenocyte cell suspension of pooled mice immediately prepared and dead cells and red blood cells were removed by Histopacque gradient separation method and freshly isolated splenocytes containing >99% viable cells were transferred (5×106, i.p) into NOD.Scid mice. Blood glucose in recipient mice was monitored at least once a week as described earlier (*: p = 0.02, Kaplan-Meier test).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3351395&req=5

pone-0036079-g003: In vivo AG490 treated splenocytes did not transfer diabetes to NOD.Scid mice.Prediabetic NOD mice (4- weeks of age) were treated with AG490 or DMSO for five consecutive week (n = 5 per group), three times per week and mice were sacrificed one week after the last injection at week 10. Splenocyte cell suspension of pooled mice immediately prepared and dead cells and red blood cells were removed by Histopacque gradient separation method and freshly isolated splenocytes containing >99% viable cells were transferred (5×106, i.p) into NOD.Scid mice. Blood glucose in recipient mice was monitored at least once a week as described earlier (*: p = 0.02, Kaplan-Meier test).
Mentions: In order to understand how AG490 reversed and blocked the onset of autoimmune diabetes in vivo, we treated prediabetic NOD mice (4-week of age) with AG490 or with DMSO for 5 week. Mice were sacrificied at week 10 and splenocyte cell suspensions were prepared with Histopaque gradient separation. The dead cells and reticulocytes were removed by the Histopaque gradient separation, therefore, dead cells were excluded without magnetic purification and without activation of T-cells and cell viability was greater than 99%. Five million freshely prepared of pooled splenocytes of NOD mice treated with AG490 (n = 5) or DMSO (n = 5) were transferred by i.p injection into recipient NOD.Scid mice and blood glucose and body weight was closely monitored. Our data as shown in figure 3 suggests that prevention of autoimmune diabetes by AG490 could be due to inactivation of atuoreactive T-cells and/or induction/generation of regulatory T-cell or other regulatory population.

Bottom Line: However, the therapeutic effects of AG490 on the development of T1D are unknown.AG490 significantly inhibited the development of T1D (p = 0.02, p = 0.005; at two different time points).The use of such agents, given their extensive safety profiles, provides a strong foundation for their translation to humans with or at increased risk for the disease.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacotherapy & Translational Research, College of Pharmacy, University of Florida, Gainesville, Florida, United States of America. dsemiromi@cop.ufl.edu

ABSTRACT

Background: Recent studies in the NOD (non-obese diabetic) mouse model of type 1 diabetes (T1D) support the notion that tyrosine kinase inhibitors have the potential for modulating disease development. However, the therapeutic effects of AG490 on the development of T1D are unknown.

Materials and methods: Female NOD mice were treated with AG490 (i.p, 1 mg/mouse) or DMSO starting at either 4 or 8 week of age, for five consecutive week, then once per week for 5 additional week. Analyses for the development and/or reversal of diabetes, insulitis, adoptive transfer, and other mechanistic studies were performed.

Results: AG490 significantly inhibited the development of T1D (p = 0.02, p = 0.005; at two different time points). Monotherapy of newly diagnosed diabetic NOD mice with AG490 markedly resulted in disease remission in treated animals (n = 23) in comparision to the absolute inability (0%; 0/10, p = 0.003, Log-rank test) of DMSO and sustained eugluycemia was maintained for several months following drug withdrawal. Interestingly, adoptive transfer of splenocytes from AG490 treated NOD mice failed to transfer diabetes to recipient NOD.Scid mice. CD4 T-cells as well as bone marrow derived dendritic cells (BMDCs) from AG490 treated mice, showed higher expression of Foxp3 (p<0.004) and lower expression of co-stimulatory molecules, respectively. Screening of the mouse immune response gene arrary indicates that expression of costimulaotry molecule Ctla4 was upregulated in CD4+ T-cell in NOD mice treated with AG490, suggesting that AG490 is not a negative regulator of the immune system.

Conclusion: The use of such agents, given their extensive safety profiles, provides a strong foundation for their translation to humans with or at increased risk for the disease.

Show MeSH
Related in: MedlinePlus