mRNA sequencing of human osteoarthritis chondrocytes treated with or without autotaxin inhibitor

OBJECTIVE: To identify the changes in gene expression elicted by ATX inhibition in human OA primary chondrocytes. Chondrocytes were obtained from patients undergoing knee artheroplasty, cultured, and treated for 24 h with ATX inhibitor or control (without inhibitor). Total RNA was then extracted, and mRNA libraries were prepared using Illumina's TruSEQ stranded mRNA library preparation kit, pooled together, and sequenced on Illumina's NextSeq550. Differential expression analysis was performed on the mRNA-SEQ data obtained from 6 prepared libraries, where 3 were treated with the ATX inhibitor. Subsequent pathway analysis identified pathways related to cholesterol analogue metabolism and cell-cycle that could be modulated by ATX inhibition in human OA chondrocytes.

研究目的：探究人骨关节炎（Osteoarthritis, OA）原代软骨细胞经自分泌运动因子（Autotaxin, ATX）抑制后，其基因表达的变化情况。研究人员从接受膝关节置换术的患者体内分离得到软骨细胞，经体外培养后，分别以ATX抑制剂或不含抑制剂的对照体系处理24小时。随后提取总RNA，使用Illumina的TruSEQ链特异性mRNA文库制备试剂盒构建mRNA文库，将所有文库混合后，在Illumina NextSeq550测序平台上进行测序。本研究针对6个构建完成的mRNA文库所获得的测序数据开展差异表达分析，其中3个文库的软骨细胞经ATX抑制剂处理。后续通路分析显示，在人骨关节炎软骨细胞中，ATX抑制可调控胆固醇类似物代谢与细胞周期相关的信号通路。