2020 Fiscal Year Annual Research Report
免疫疾患におけるＲＮＡ分解酵素Ｒｅｇｎａｓｅ－１調節機構解明とその操作法の開発
| Project/Area Number |
19J23450
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| Research Institution | Kyoto University |
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Principal Investigator |
Tse Ka Man Carman 京都大学, 医学研究科, 特別研究員(DC1)
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| Project Period (FY) |
2019-04-25 – 2022-03-31
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| Keywords | Regnase-1 / autoimmune diseases / pulmonary diseases / mRNA stability / antisense oligos |
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| Outline of Annual Research Achievements |
It was reported that mice with Regnase-1 deficiency spontaneously develop lymphadenopathy and splenomegaly; while Regnase-1 haploinsuficient mice display exacerbated inflammation in autoimmune disease and pulmonary inflammation models, therefore we hypothesized that enhancing the availability of Regnase-1 protein could relieve the inflammatory responses and shifting the balance in favor of an anti-inflammatory state.
We found that targeting the secondary structural elements, i.e. the stem-loop structures, located on the Regnase-1 3'UTR could effectively reduce LPS-mediated pro-inflammatory cytokine production such as Il6, Il1b and Tnf in purified bone marrow-derived macrophages; and in vivo using various pulmonary disease and autoimmune disease models. To our knowledge, this is the first research project showing that manipulation of Regnase-1 availability can attenuate disease severity in various inflammatory disease models and represent a new strategy to influence inflammatory and autoimmune responses in a therapeutic setting.
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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
This year we evaluated the safety and therapeutic aspects of MO in various primary cells (in vitro) and mouse models (in vivo).
We successfully demonstrated that enhanced Regnase-1 availability by morpholino oligonucleotides exerted therapeutic effects in LPS-lung inflammation model by reducing the expression of inflammatory cytokines as well as alveolar neutrophil accumulation in lung tissue and bronchoalveolar lavage fluid. Furthermore, treatment with MO markedly alleviated the histopathology changes induced by LPS, most significantly by retaining the pulmonary interstitial spaces and reducing the hemorrhage incidence.
As an alternative model, we also tested the therapeutic potential of Regnase-1 in treating autoimmune diseases such as experimental autoimmune encephalitis model. We observed that MO treatment effectively postponed and relived the EAE symptoms and allowed better weight recovery, which was associated with a significant reduction of spinal cord/brain-infiltrating immune cells.
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| Strategy for Future Research Activity |
In the following year, we will identify the cell types and physiologically relevant pathways that were altered by increased Regnase-1 abundance by performing transcriptome analysis and single cell RNA sequencing in the disease models tested.
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