[P2-2-03] Identification and functional analysis of genes involved in the onset and progression of osteoarthritis
Keywords:変形性関節症、マウスジェネティクス、ゲノム編集
[Objective] Osteoarthritis (OA) is a prevalent irreversible degradation of joint disease. Because molecular mechanisms involved in pathogenesis of OA are still elusive, there are no effective therapy for the patients. To develop early diagnostic markers and effective therapy of OA, we attempted to identify the genes associated with pathogenesis of OA. [Methods] The destabilization of the medial meniscus (DMM) treatment and sham operations were performed in right and left knee joints of 8 weeks C57BL/6J male mice, respectively. 4 weeks after the treatments, the femoral and tibial heads were isolated from the mice, and total RNA extracted from the articular cartilage tissues was subjected to RNA-seq and RT-qPCR analyses. [Results] According to the previous studies that Col10a1 expression is increased by DMM treatment, we subjected the 2 samples in which Col10a1 expressions were elevated in the DMM sides compared to the sham sides to the RNA-seq analyses. The RNA-seq analyses indicated upregulation of 193 genes in DMM sides in comparison with the sham sides among approximately 20000 genes. RT-qPCR analyses suggest that 4 candidate genes including Dpt elevated on the DMM side may be involved in the pathogenesis of OA. We succeeded to generate 4 genes mutant mice by CRISPR/Cas9-based TAKE method. [Discussion] We could identify the candidate genes that would be involved in onset and/or progression of OA. We succeeded to generate knockout mice of these candidate genes associated with OA. We are plannning to analyze the pathogenic roles of the genes after DMM treatment in the mice.