2013 Fiscal Year Annual Research Report
| Project/Area Number |
12F02736
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| Research Institution | Osaka University |
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Principal Investigator |
審良 静男 大阪大学, 免疫学フロンティア研究センター, 教授
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| Co-Investigator(Kenkyū-buntansha) |
KEBER Rok 大阪大学, 免疫学フロンティア研究センター, 外国人特別研究員
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| Keywords | Renase-1 / RNA stabilit |
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| Research Abstract |
Primary goal of the FY2013 was to characterize immune cell-specific targets of Regnase-1, which is currently one of the main interests of my hosting laboratory. Regnase-1 is an enzyme critical for the suppression of autoimmunity by controlling the stability of mRNAs that encode immune-related factors. Since the discovery of immunomodulatory function of Regnase-1 in 2009, there have been three high impact publications, describing targets and function of this protein in macrophages and CD4 subset of T-cells. My aim is to examine the specific modes of action in additional two immune cell types ; cytotoxic T cells (CD8) and regulatory T-cells (Treg) and reveal novel and specific targets of regnase. Outline of the research performance can be summarized in the following points ;
1. Development of conditional mouse models to study CD8 and Treg-specific role or Regnase-1
2. Extensive phenotypisation of immune parameters in both animal models
3. Functional characterization of CD8 and Treg cells
3. In depth analysis of gene expression in naïve and stimulated CD8 and Treg cells with ablated Regnase-1 protein
In parallel to Regnase-1 project, I aim to perform in vivo studies of Myd88 mutations with oncogenic potential. I designed and generated a knock-in DNA construct that will enable a targeted replacement of the wild-type Myd86 allele with orthologous-human L265P-carrying allele in the mouse. This mutation has been described in some types of human lymphoma, but the mechanism has not been studied in detail. In addition, potential gain-of-function Myd88 mutants obtained by random ENU mutagenesis in RIKEN center (Japan), will be examined.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
Characterization of cell-specific in vivo role of Regnase-1 in two immune cell types is a laborious and time consuming work, especially when performed individually (not in a bigger collaborative research group). Despite the fact that all the research is going according to the plan, publications (high quality) are to be expected only by the end of the tenure.
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| Strategy for Future Research Activity |
Further plan in FY2014 is to finish high throughput transcriptional analysis of Treg cells with Regnase-1 disfunction and identify novel cell-specific targets of Regnase-1. After that most interesting targets will be examined in detailed using gene knockouts, lentiviral gene silencing and overexpression. Main aim is to dissect in details the Regnase-1-affected downstream pathways and reveal previously unidentified molecular mechanisms leading to the onset of autoimmune phenotype.
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