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Abstract：
OBJECTIVE: To characterize aspects of triiodothyronine (T3) induced chondrocyte terminal maturation within the molecular osteoarthritis pathophysiology using the previously established T3 human ex vivo osteochondral explant model.
METHODS: RNA-sequencing was performed on explant cartilage obtained from OA patients (n = 8), that was cultured ex vivo with or without T3 (10 ng/ml), and main findings were validated using RT-qPCR in an independent sample set (n = 22). Enrichment analysis was used for functional clustering and comparisons with available OA patient RNA-sequencing and GWAS datasets were used to establish relevance for OA pathophysiology by linking to OA patient genomic profiles.
RESULTS: Besides the upregulation of known hypertrophic genes EPAS1 and ANKH, T3 treatment resulted in differential expression of 247 genes with main pathways linked to extracellular matrix and ossification. CCDC80, CDON, ANKH and ATOH8 were among the genes found to consistently mark early, ongoing and terminal maturational OA processes in patients. Furthermore, among the 37 OA risk genes that were significantly affected in cartilage by T3 were COL12A1, TNC, SPARC and PAPPA.
CONCLUSIONS: RNA-sequencing results show that metabolic activation and recuperation of growth plate morphology are induced by T3 in OA chondrocytes, indicating terminal maturation is accelerated. The molecular mechanisms involved in hypertrophy were linked to all stages of OA pathophysiology and will be used to validate disease models for drug testing.
METHODS: RNA-sequencing was performed on explant cartilage obtained from OA patients (n = 8), that was cultured ex vivo with or without T3 (10 ng/ml), and main findings were validated using RT-qPCR in an independent sample set (n = 22). Enrichment analysis was used for functional clustering and comparisons with available OA patient RNA-sequencing and GWAS datasets were used to establish relevance for OA pathophysiology by linking to OA patient genomic profiles.
RESULTS: Besides the upregulation of known hypertrophic genes EPAS1 and ANKH, T3 treatment resulted in differential expression of 247 genes with main pathways linked to extracellular matrix and ossification. CCDC80, CDON, ANKH and ATOH8 were among the genes found to consistently mark early, ongoing and terminal maturational OA processes in patients. Furthermore, among the 37 OA risk genes that were significantly affected in cartilage by T3 were COL12A1, TNC, SPARC and PAPPA.
CONCLUSIONS: RNA-sequencing results show that metabolic activation and recuperation of growth plate morphology are induced by T3 in OA chondrocytes, indicating terminal maturation is accelerated. The molecular mechanisms involved in hypertrophy were linked to all stages of OA pathophysiology and will be used to validate disease models for drug testing.
摘要：
目的：使用先前建立的T3人离体骨软骨外植体模型，在分子骨关节炎病理生理学中表征三碘甲状腺原氨酸（T3）诱导的软骨细胞终末成熟的方面。
方法：对从OA患者（n=8）获得的外植体软骨进行RNA测序，在有或没有T3（10ng/ml）的情况下离体培养，和主要发现在一个独立的样品集（n=22）中使用RT-qPCR进行验证。富集分析用于功能聚类，并与可用的OA患者RNA测序和GWAS数据集进行比较，通过与OA患者基因组谱链接，建立与OA病理生理学的相关性。
结果：除了已知的肥大基因EPAS1和ANKH的上调，T3处理导致247个基因的差异表达，其主要途径与细胞外基质和骨化有关。CCDC80,CDON,ANKH和ATOH8是早期发现一致标记的基因之一，患者正在进行和终末成熟的OA过程。此外，在37个OA风险基因中，T3显著影响软骨的是COL12A1，TNC，SPARC和PAPPA。
结论：RNA测序结果表明，T3诱导OA软骨细胞代谢激活和生长板形态恢复，表明终端成熟加速。肥大中涉及的分子机制与OA病理生理学的所有阶段相关，并将用于验证药物测试的疾病模型。
方法：对从OA患者（n=8）获得的外植体软骨进行RNA测序，在有或没有T3（10ng/ml）的情况下离体培养，和主要发现在一个独立的样品集（n=22）中使用RT-qPCR进行验证。富集分析用于功能聚类，并与可用的OA患者RNA测序和GWAS数据集进行比较，通过与OA患者基因组谱链接，建立与OA病理生理学的相关性。
结果：除了已知的肥大基因EPAS1和ANKH的上调，T3处理导致247个基因的差异表达，其主要途径与细胞外基质和骨化有关。CCDC80,CDON,ANKH和ATOH8是早期发现一致标记的基因之一，患者正在进行和终末成熟的OA过程。此外，在37个OA风险基因中，T3显著影响软骨的是COL12A1，TNC，SPARC和PAPPA。
结论：RNA测序结果表明，T3诱导OA软骨细胞代谢激活和生长板形态恢复，表明终端成熟加速。肥大中涉及的分子机制与OA病理生理学的所有阶段相关，并将用于验证药物测试的疾病模型。
