RNA Sequencing Reveals LINC00167 as a Potential Diagnosis Biomarker for Primary Osteoarthritis: A Multi-Stage Study

Liying Jiang; Yiqin Zhou; Junjie Shen; Yi Chen; Ziyuan Ma; Yuhui Yu; Minjie Chu; Qirong Qian; Xun Zhuang; Shengli Xia

doi:10.3389/fgene.2020.539489

RNA Sequencing Reveals LINC00167 as a Potential Diagnosis Biomarker for Primary Osteoarthritis: A Multi-Stage Study

Front Genet. 2021 Jan 14:11:539489. doi: 10.3389/fgene.2020.539489. eCollection 2020.

Authors

Liying Jiang¹, Yiqin Zhou², Junjie Shen³, Yi Chen², Ziyuan Ma⁴, Yuhui Yu³, Minjie Chu³, Qirong Qian¹, Xun Zhuang³, Shengli Xia⁵

Affiliations

¹ Shanghai Key Laboratory for Molecular Imaging, Shanghai University of Medicine & Health Sciences, Shanghai, China.
² Department of Joint Surgery and Sports Medicine, Shanghai Changzheng Hospital, Second Military Medical University, Huangpu, China.
³ Department of Epidemiology, School of Public Health, Nantong University, Nantong, China.
⁴ Department of Orthopedics, School of Medical, Strategically Strategic Medical University, Guiyang, China.
⁵ Department of Orthopedics, Shanghai University of Medicine & Health Sciences Affiliated Zhoupu Hospital, Shanghai, China.

Abstract

Objectives: Given the roles played by lncRNA in human diseases and the high incidence of OA, this study investigated the pivotal pathways involved in the disease and identified potential biomarkers for OA diagnosis.

Methods: We first performed an exploration of RNA-sequencing in peripheral blood leukocytes from six subjects (3 OA and 3 healthy controls). Promising candidate lncRNAs were evaluated in first stage validation using a GEO dataset (GSE114007) of 38 subjects (20 OA and 18 healthy controls), followed by a second stage validation using quantitative PCR analysis with 101 subjects (67 OA and 34 controls). The third stage investigated the potential value of validated lncRNA in the early diagnosis of OA in peripheral blood leukocytes from a total of 120 participants (60 cases and 60 controls).

Results: The dataset identified a total of 1,380 up-regulated and 719 down-regulated mRNAs and 5,743 up-regulated and 7,384 down-regulated lncRNAs. The up-regulated DEGs were mainly enriched in the extracellular matrix, while the down-regulated DEGs were mainly enriched in the IL-17 and wnt signaling pathways. 18 overlapping candidate lncRNAs survived after first-stage validation. 3 hub lncRNAs were selected for the second validation stage and qualified in an external sample, and lncRNA LINC00167 was further confirmed with a similar result (down-expressed in both stages). Receiver operating characteristic analysis showed that LINC00167 can distinguish OA cases from healthy controls with a high area under the curve of 0.879 (95%CI: 0.819, 0.938; P < 0.001), with a sensitivity of 80.7% and specificity of 83.5%.

Conclusion: The expression profile of OA was identified and critical pathways were elucidated by an integrated approach to RNA-seq from easily accessible blood. LINC00167 may serve as a potential early diagnosis marker for OA in clinical practice. The detailed mechanism of action of this lncRNA requires further elucidation in future studies.

Keywords: RNA-seq; lncRNA; marker; osteoarthritis; peripheral blood leukocytes.