2022 Fiscal Year Research-status Report
Elucidating the role of IL-1R2, a decoy receptor of IL-1, in inflammation-induced stem cell aging
| Project/Area Number |
22K15126
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| Research Institution | Kumamoto University |
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Principal Investigator |
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| Project Period (FY) |
2022-04-01 – 2025-03-31
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| Keywords | IL-1R2 / TPA / Tet-on system |
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| Outline of Annual Research Achievements |
Experimental Plan1: We successfully generated Il1r2CreER/Rosa-tdTomato lines. Tamoxifen-treated Il1r2CreER/Rosa-tdTomato mice, either through intraperitoneal injection (IP) or diet, exhibited tomato signal specifically in the inter-scale region, a typical territory of slow-cycling stem cells in the mouse tail skin.
The lineage tracing of these tomato-positive basal cells in homeostasis or IL1a/b-induced condition will reveal their stem cell potential and behavior.
Experimental Plan 2: Using the Tet-on system, we successfully generated an Il1r2 over-expressing mouse line. Il1r2 over-expressing mice counteracted the effect of TPA, an IL1a-mediated inflammatory agent, on tail skin compared to wild-type control mice.
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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
It takes time for mouse mating and housing.
I had a problem with the in vitro experiment since I needed to spend time to do troubleshooting the primary keratinocytes isolation protocol at Kumamoto University.
The troubleshooting has already finished, and I could be able to isolate primary keratinocytes from mice so we can proceed with the in vitro experiment.
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| Strategy for Future Research Activity |
Experimental Plan 1:The lineage tracing experiment using Il1r2CreER/Rosa-tdTomato lines under homeostasis or inflammatory environment will be conducted
Experimental Plan 2: Il1r2 conditionally knock-out mice are being generated.
Experimental Plan 3: To investigate the molecular mechanisms by which IL-1R2 protects epidermal stem cells (SCs) from chronic inflammation, mouse primary keratinocytes, either control or IL-1R2 over-expressing cells, will be treated with IL-1a/b in vitro, then the activation of canonical downstream molecules of IL-1 signaling pathways including NFkB, MAPK (JNK1/2/3, ERK1/2) will be tested. The impact of IL-1a/b exposure on cellular phenotypes, including cell proliferation, survival, differentiation, ROS production, and SC marker expressions, will be investigated.
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| Causes of Carryover |
Because of Covid19 situation, some reagents and antibodies did not arrive on time
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