| 研究実績の概要 |
Collagen-induced arthritis is performed and arthritic synovial cells are recovered from the inflamed synovium (knee and ankle joints) by collagenase digestion and analyzed by FACS. We used the following surface protein antibodies to distinguish the different cell populations in arthritic joints. In arthritic condition, immune cells accumulate in the inflamed synovium, the majority of which are CD45+CD11b+ myeloid cells, and also the CD45+CD3+ T lymphocytes. The hypertrophic synovium is also enriched in the CD45-negative mesenchymal lineage cells, of which the majority is FLSs, which are defined by the lack of CD45 and characteristic expression of surface protein podoplanin (gp38), but also endothelial cells (CD45-CD31+) and pericytes (CD45-CD146+).
In order to clarify the major osteoclastogenic factor-expressing cells in arthritic joints, we FACS-sorted several cells from fresh arthritic synovium and examined for the expression of osteoclastogenic factors using our newly developed reporter mice. We found that the majority of the cells that express osteoclastogenic factors within the inflamed synovium are FLSs (CD45-gp38+), but not other cell populations. Hence, our preliminary results indicate that FLSs are the major osteoclastogenic cells in arthritic joints.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
Arthritis model was successfully induced and arthritic synovial cells were isolated from the inflamed synovium (knee and ankle joints) by enzymatic digestion. We performed FACS analysis and identified different cell populations in arthritic joints. In arthritic condition, immune cells accumulated in the inflamed synovium, and also be enriched in the CD45-negative mesenchymal lineage cells, of which the majority were FLSs. Importantly, we found that the majority of the cells that express osteoclastogenic factors are FLSs. Hence, our preliminary results accord with our experimental expectations.
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| 今後の研究の推進方策 |
To identify the unique characteristics of bone-damaging FLSs, we plan to perform the RNAseq analysis. Differential expression analysis and Gene set enrichment analysis will be performed to define the molecular and gene signatures of bone-damaging FLSs. In addition, pathway and gene-ontology enrichment analysis will be performed to define the candidate pathways that contribute to the induction of bone-damaging FLSs. We will also generate the osteoclastogenic factor-deficient mice to evaluate the functions in vivo under the arthritic condition.
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