Long Intergenic Noncoding RNAs Mediate the Human Chondrocyte Inflammatory Response and Are Differentially Expressed in Osteoarthritis Cartilage.
Long Intergenic Noncoding RNAs Mediate the Human Chondrocyte Inflammatory Response and Are Differentially Expressed in Osteoarthritis Cartilage.
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DOI：
10.1002/art.39520
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发表时间：
2016-04
期刊：
影响因子：
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通讯作者：
Jones SW
中科院分区：
文献类型：
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作者：
Pearson MJ;Philp AM;Heward JA;Roux BT;Walsh DA;Davis ET;Lindsay MA;Jones SW
鉴定与人原发性骨关节炎（OA）软骨细胞炎症反应相关的长链非编码RNA（lncRNA），包括长链基因间非编码RNA（lincRNA）、反义RNA和假基因，并探讨其在OA中的表达和功能。OA软骨从关节置换手术后的髋关节或膝关节OA患者获得。非OA软骨来自死后供体和股骨颈骨折患者。胶原酶消化分离原代OA软骨细胞。通过RNA测序（RNAseq）和定量逆转录酶-聚合酶链反应进行LncRNA表达分析。使用LNA longRNA GapmeR（Lnqon）实现lncRNA软骨细胞表达的调节。用Luminex测量细胞因子产生。RNaseq在原代人髋关节OA软骨细胞中鉴定出983个lncRNA，其中183个之前未被鉴定。在白细胞介素-1 β（IL-1β）刺激后，我们鉴定了125种差异表达的lincRNA。与非OA软骨相比，lincRNA p50相关的环氧合酶2基因外RNA（PACER）和2种新型软骨细胞炎症相关lincRNA（CILinc 01和CILinc 02）在膝关节和髋关节OA软骨中的表达差异。在原代OA软骨细胞中，这些lincRNA响应于多种促炎细胞因子而被快速且瞬时地诱导。人软骨细胞中CILinc 01和CILinc 02表达的敲低显著增强了IL-1刺激的促炎细胞因子的分泌。人OA软骨细胞中的炎症反应与lncRNA谱的广泛变化相关，包括PACER、CILinc 01和CILinc 02。髋关节和膝关节OA软骨中CILinc 01和CILinc 02的差异表达及其在软骨细胞炎症反应期间调节细胞因子产生的作用表明，它们可能在介导OA中炎症驱动的软骨退变中发挥重要作用。
To identify long noncoding RNAs (lncRNAs), including long intergenic noncoding RNAs (lincRNAs), antisense RNAs, and pseudogenes, associated with the inflammatory response in human primary osteoarthritis (OA) chondrocytes and to explore their expression and function in OA. OA cartilage was obtained from patients with hip or knee OA following joint replacement surgery. Non‐OA cartilage was obtained from postmortem donors and patients with fracture of the neck of the femur. Primary OA chondrocytes were isolated by collagenase digestion. LncRNA expression analysis was performed by RNA sequencing (RNAseq) and quantitative reverse transcriptase–polymerase chain reaction. Modulation of lncRNA chondrocyte expression was achieved using LNA longRNA GapmeRs (Exiqon). Cytokine production was measured with Luminex. RNAseq identified 983 lncRNAs in primary human hip OA chondrocytes, 183 of which had not previously been identified. Following interleukin‐1β (IL‐1β) stimulation, we identified 125 lincRNAs that were differentially expressed. The lincRNA p50‐associated cyclooxygenase 2–extragenic RNA (PACER) and 2 novel chondrocyte inflammation–associated lincRNAs (CILinc01 and CILinc02) were differentially expressed in both knee and hip OA cartilage compared to non‐OA cartilage. In primary OA chondrocytes, these lincRNAs were rapidly and transiently induced in response to multiple proinflammatory cytokines. Knockdown of CILinc01 and CILinc02 expression in human chondrocytes significantly enhanced the IL‐1–stimulated secretion of proinflammatory cytokines. The inflammatory response in human OA chondrocytes is associated with widespread changes in the profile of lncRNAs, including PACER, CILinc01, and CILinc02. Differential expression of CILinc01 and CIinc02 in hip and knee OA cartilage, and their role in modulating cytokine production during the chondrocyte inflammatory response, suggest that they may play an important role in mediating inflammation‐driven cartilage degeneration in OA.