Next generation sequencing identifies miRNA-based biomarker panel for lupus nephritis

// Yu-Jih Su 1 , I-Chun Lin 2 , Lin Wang 3 , Cheng-Hsien Lu 4,5,6 , Yi-Ling Huang 1 and Ho-Chang Kuo 7 1 Department of Rheumatology, Allergy and Immunology, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung, Taiwan 2 Department of Pediatrics, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung, Taiwan 3 Department of Pediatrics, PoJen Hospital, Kaohsiung, Taiwan 4 Department of Neurology, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung, Taiwan 5 Department of Biological Science, National Sun Yat-Sen University, Kaohsiung, Taiwan 6 Department of Neurology, Xiamen Chang Gung Memorial Hospital, Xiamen, Fujian, China 7 Department of Pediatrics and Kawasaki Disease Center, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, Taiwan Correspondence to: Ho-Chang Kuo, email: erickuo48@yahoo.com.tw Keywords : next generation sequencing; lupus; nephritis; microRNA; Immunology Received: October 29, 2017     Accepted: May 08, 2018     Published: June 15, 2018 Abstract The symptomatology of lupus nephritis (LN) consists of foamy urine and lower leg edema, as well as such systemic manifestations as oral ulcers, arthralgia/arthritis, and lymphadenopathy. However, these symptoms may appear mild and non-specific. If these symptoms are unrecognized, thus delaying treatment, approximately 10% of LN patients will develop permanent kidney damage and end-stage kidney disease. Therefore, the purpose of this study is to identify a surrogate biomarker for the early detection of LN. In this study, we first adopted next generation sequencing (NGS) in order to screen differential expression levels of microRNA between SLE patients with and without LN. The results of both the NGS and the literature review confirmed the potential of 15 microRNAs through real-time qPCR. We further considered clinical laboratory data for additional analysis. In total, 41 microRNAs demonstrated significant differences through NGS screening. We then verified eight microRNAs from NGS and seven microRNAs from the literature review using the real-time qPCR method in peripheral mononuclear cells. Ultimately, mir-125a-5p, miR-146a-5p, and mir-221-3p were found to be statistically significant not only in the screening study but also in the real-time qPCR verification studies. miR-146a-5p was observed to have a significant correlation with clinical biochemistry markers, as well as to be a surrogate biomarker for the early detection of lupus nephritis. This study is the first to show that the intracellular biomarker miR-146a-5p may serve as a useful specific biomarker for the detection of lupus nephritis among lupus patients in the future, regardless of serum albumin levels and spot urine protein/creatinine ratio.