2020 Fiscal Year Research-status Report
The role of the transcription factor Tox2 in Treg and Tfh biology
| Project/Area Number |
18K07175
|
|---|
| Research Institution | Osaka University |
|---|
Principal Investigator |
ウィング ジェイムス 大阪大学, 免疫学フロンティア研究センター, 特任准教授(常勤) (00648694)
|
|---|
| Project Period (FY) |
2018-04-01 – 2022-03-31
|
| Keywords | Regulatory T-cells / T-follicular regulatory / T-follicular helper / Mass Cytometry (CyTOF) |
|---|
| Outline of Annual Research Achievements |
We have now bred the Tox2 flox mice to Foxp3-IRES-Cre mice and found some evidence of disruption of the germinal center environment and particularly IgA producing B-cells in the payer’s patch of the gut environment. This suggests that Tox2 expressing Tregs and particualrly Tfr may have their function regulated by Tox2.
Since Tox2 appears to be upregulated by TCR stimulation in vitro we have used CRISPR KO of around 50 genes in human cells to determine the effect of components of TCR signaling pathways on the expression of Tox2 in CD4 T-cells. Here we find that particularly modulation of the tumor necrosis factor alpha-induced protein 3 (TNFAIP3/A20) a negative regulator of TCR mediated Nf-KB function. The expression of Tox2 correlates with expression of it family member Tox to a certain extent but does not have an identical expression profile.
|
| Current Status of Research Progress |
Current Status of Research Progress
3: Progress in research has been slightly delayed.
Reason
We did have some disruption of our plans due to the COVID-19 outbreak. In particular this affected staff recruitment due to the difficulty of international travel. As a result, we requested some carry over of funding into a fourth year.
|
| Strategy for Future Research Activity |
Our plan for the next year is more detailed analysis of the effect of the loss of Tox2 on murine Tregs by vaccination experiments. In these experiments we will vaccinate mice with NP-Ova allowing the detailed measurement of changes to antibody responses and germinal center formation. We will also closely monitor the formation of antigen specific plasma cells since recent accumulating evidence is that Tfr have a particularly important role in plasma cell formation.
We have also further refined our Mass cytometry technique to allow the monitoring of the immunometabolism and will use this to define any effects of Tox2 on metabolic parameter of Tregs. Tox is known to have some role in the control of the immunometabolism so we also expect some effect from Tox2.
Additionally, we will monitor the effect of Tox2 CRISPR on human cells particularly in the context of changes to the immunometabolism by mass cytometry and chromatin accessibility as measured by single-cell Assay for Transposase Accessible Chromatin by sequencing (scATAC-seq) a novel form of sequencing that we recently contributed too the invention of.
|
| Causes of Carryover |
We did have some disruption of our plans due to the COVID-19 outbreak. As a result, we requested some carry over of funding into a fourth year. The research plan is not substantially altered by this.
|
