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The kinase DYRK1A reciprocally regulates the differentiation of Th17 and regulatory T cells.

Khor B, Gagnon JD, Goel G, Roche MI, Conway KL, Tran K, Aldrich LN, Sundberg TB, Paterson AM, Mordecai S, Dombkowski D, Schirmer M, Tan PH, Bhan AK, Roychoudhuri R, Restifo NP, O'Shea JJ, Medoff BD, Shamji AF, Schreiber SL, Sharpe AH, Shaw SY, Xavier RJ - Elife (2015)

Bottom Line: Importantly, both Treg cells generated using the DYRK1A inhibitor harmine and direct administration of harmine itself potently attenuate inflammation in multiple experimental models of systemic autoimmunity and mucosal inflammation.Our results identify DYRK1A as a physiologically relevant regulator of Treg cell differentiation and suggest a broader role for other DYRK family members in immune homeostasis.These results are discussed in the context of human diseases associated with dysregulated DYRK activity.

View Article: PubMed Central - PubMed

Affiliation: Gastrointestinal Unit and Center for the Study of Inflammatory Bowel Disease, Massachusetts General Hospital, Harvard Medical School, Boston, United States.

ABSTRACT
The balance between Th17 and T regulatory (Treg) cells critically modulates immune homeostasis, with an inadequate Treg response contributing to inflammatory disease. Using an unbiased chemical biology approach, we identified a novel role for the dual specificity tyrosine-phosphorylation-regulated kinase DYRK1A in regulating this balance. Inhibition of DYRK1A enhances Treg differentiation and impairs Th17 differentiation without affecting known pathways of Treg/Th17 differentiation. Thus, DYRK1A represents a novel mechanistic node at the branch point between commitment to either Treg or Th17 lineages. Importantly, both Treg cells generated using the DYRK1A inhibitor harmine and direct administration of harmine itself potently attenuate inflammation in multiple experimental models of systemic autoimmunity and mucosal inflammation. Our results identify DYRK1A as a physiologically relevant regulator of Treg cell differentiation and suggest a broader role for other DYRK family members in immune homeostasis. These results are discussed in the context of human diseases associated with dysregulated DYRK activity.

No MeSH data available.


Related in: MedlinePlus

Effect of harmine on NFAT1 nuclear localization with time.Time-course Amnis analyses showing effect of harmine (blue) relative to control Treglow conditions (black).DOI:http://dx.doi.org/10.7554/eLife.05920.022
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fig4s1: Effect of harmine on NFAT1 nuclear localization with time.Time-course Amnis analyses showing effect of harmine (blue) relative to control Treglow conditions (black).DOI:http://dx.doi.org/10.7554/eLife.05920.022

Mentions: DYRKs phosphorylate several proteins (Aranda et al., 2011). Notable amongst these in the context of T cell biology are NFAT proteins, whose phosphorylation by DYRKs leads to their nuclear exclusion (Gwack et al., 2006). Thus, inhibition of DYRKs would be predicted to lead to increased levels of NFAT in the nucleus. To assess this hypothesis, we performed studies using Amnis technology, which combines flow cytometry and high-resolution microscopy to allow precise quantitation of intracellular localization of individual proteins. Naïve CD4+ T cells largely retain NFAT1 in the cytoplasm (Figure 4I, black line). Upon stimulation in Treglow conditions, nuclear translocation of NFAT1 is observed with an accompanying right shift of the nuclear/cytoplasmic ratio (Figure 4I, red line). This nuclear translocation of NFAT1 is reproducibly enhanced approximately 40% with the addition of harmine (Figure 4I, blue line). In support of these results, we independently fractionated nuclei from cells treated with each of these conditions and performed Western blot analyses. These also showed increased NFAT1 in the nuclear compartment upon stimulation in Treglow conditions, with a similar (40%) additional increase when harmine was added (Figure 4J). Thus, harmine enhances nuclear accumulation of NFAT1 at early time points up to 2 hr after stimulation (Figure 4—figure supplement 1).


The kinase DYRK1A reciprocally regulates the differentiation of Th17 and regulatory T cells.

Khor B, Gagnon JD, Goel G, Roche MI, Conway KL, Tran K, Aldrich LN, Sundberg TB, Paterson AM, Mordecai S, Dombkowski D, Schirmer M, Tan PH, Bhan AK, Roychoudhuri R, Restifo NP, O'Shea JJ, Medoff BD, Shamji AF, Schreiber SL, Sharpe AH, Shaw SY, Xavier RJ - Elife (2015)

Effect of harmine on NFAT1 nuclear localization with time.Time-course Amnis analyses showing effect of harmine (blue) relative to control Treglow conditions (black).DOI:http://dx.doi.org/10.7554/eLife.05920.022
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4441007&req=5

fig4s1: Effect of harmine on NFAT1 nuclear localization with time.Time-course Amnis analyses showing effect of harmine (blue) relative to control Treglow conditions (black).DOI:http://dx.doi.org/10.7554/eLife.05920.022
Mentions: DYRKs phosphorylate several proteins (Aranda et al., 2011). Notable amongst these in the context of T cell biology are NFAT proteins, whose phosphorylation by DYRKs leads to their nuclear exclusion (Gwack et al., 2006). Thus, inhibition of DYRKs would be predicted to lead to increased levels of NFAT in the nucleus. To assess this hypothesis, we performed studies using Amnis technology, which combines flow cytometry and high-resolution microscopy to allow precise quantitation of intracellular localization of individual proteins. Naïve CD4+ T cells largely retain NFAT1 in the cytoplasm (Figure 4I, black line). Upon stimulation in Treglow conditions, nuclear translocation of NFAT1 is observed with an accompanying right shift of the nuclear/cytoplasmic ratio (Figure 4I, red line). This nuclear translocation of NFAT1 is reproducibly enhanced approximately 40% with the addition of harmine (Figure 4I, blue line). In support of these results, we independently fractionated nuclei from cells treated with each of these conditions and performed Western blot analyses. These also showed increased NFAT1 in the nuclear compartment upon stimulation in Treglow conditions, with a similar (40%) additional increase when harmine was added (Figure 4J). Thus, harmine enhances nuclear accumulation of NFAT1 at early time points up to 2 hr after stimulation (Figure 4—figure supplement 1).

Bottom Line: Importantly, both Treg cells generated using the DYRK1A inhibitor harmine and direct administration of harmine itself potently attenuate inflammation in multiple experimental models of systemic autoimmunity and mucosal inflammation.Our results identify DYRK1A as a physiologically relevant regulator of Treg cell differentiation and suggest a broader role for other DYRK family members in immune homeostasis.These results are discussed in the context of human diseases associated with dysregulated DYRK activity.

View Article: PubMed Central - PubMed

Affiliation: Gastrointestinal Unit and Center for the Study of Inflammatory Bowel Disease, Massachusetts General Hospital, Harvard Medical School, Boston, United States.

ABSTRACT
The balance between Th17 and T regulatory (Treg) cells critically modulates immune homeostasis, with an inadequate Treg response contributing to inflammatory disease. Using an unbiased chemical biology approach, we identified a novel role for the dual specificity tyrosine-phosphorylation-regulated kinase DYRK1A in regulating this balance. Inhibition of DYRK1A enhances Treg differentiation and impairs Th17 differentiation without affecting known pathways of Treg/Th17 differentiation. Thus, DYRK1A represents a novel mechanistic node at the branch point between commitment to either Treg or Th17 lineages. Importantly, both Treg cells generated using the DYRK1A inhibitor harmine and direct administration of harmine itself potently attenuate inflammation in multiple experimental models of systemic autoimmunity and mucosal inflammation. Our results identify DYRK1A as a physiologically relevant regulator of Treg cell differentiation and suggest a broader role for other DYRK family members in immune homeostasis. These results are discussed in the context of human diseases associated with dysregulated DYRK activity.

No MeSH data available.


Related in: MedlinePlus