Identification and validation of circulating miRNAs as potential new biomarkers for severe liver disease in patients with leptospirosis

Background Leptospirosis, a global zoonotic infectious disease, has various clinical manifestations ranging from mild self-limiting illness to life-threatening with multi-organ damage, including liver involvement. This study was aimed at identifying circulating microRNAs (miRNAs) as novel biomarkers for predicting severe liver involvement in patients with leptospirosis. Methods In a discovery set, 12 serum samples of patients with anicteric and icteric leptospirosis at initial clinical presentation were used for miRNA profiling by a NanoString nCounter miRNA assay. In a validated cohort, top candidate miRNAs were selected and further tested by qRT-PCR in serum samples of 81 and 16 individuals with anicteric and icteric leptospirosis, respectively. Results The discovery set identified 38 significantly differential expression miRNAs between the two groups. Among these, miR-601 and miR-630 were selected as the top two candidates significantly up-regulated expressed in the icteric group. The enriched KEGG pathway showed that these miRNAs were mainly involved in immune responses and inflammation. In the validated cohort, miR-601 and miR-630 levels were significantly higher in the icteric group compared with the anicteric group. Additionally, these two miRNAs displayed good predictors of subsequent acute liver failure with a high sensitivity of 100%. On regression analysis, elevated miR-601 and miR-630 expression were also predictive of multi-organ failures and poor overall survival. Conclusion Our data indicated that miRNA expression profiles were significantly differentiated between the icteric and anicteric groups. Serum miR-601 and miR-630 at presentation could potentially serve as promising biomarkers for predicting subsequent acute liver failure and overall survival in patients with leptospirosis.

[1]  Dongming Xing,et al.  Circulating microRNAs: biomarkers of disease. , 2021, Clinica chimica acta; international journal of clinical chemistry.

[2]  Bo Zhong,et al.  RNF115 plays dual roles in innate antiviral responses by catalyzing distinct ubiquitination of MAVS and MITA , 2020, Nature Communications.

[3]  N. Chuaypen,et al.  Circulating microtranscriptome profiles reveal distinct expression of microRNAs in severe leptospirosis , 2020, PLoS neglected tropical diseases.

[4]  X. Zhao,et al.  A novel circulating miRNA‐based signature for the early diagnosis and prognosis prediction of non–small‐cell lung cancer , 2020, Journal of clinical laboratory analysis.

[5]  D. Langford,et al.  MicroRNAs Regulated by the LPS/TLR2 Immune Axis as Bona Fide Biomarkers for Diagnosis of Acute Leptospirosis , 2020, mSphere.

[6]  G. Gao,et al.  Insights into catalysis and regulation of non-canonical ubiquitination and deubiquitination by bacterial deamidase effectors , 2020, Nature Communications.

[7]  B. Ganesh,et al.  Leptospirosis: a neglected tropical zoonotic infection of public health importance—an updated review , 2020, European Journal of Clinical Microbiology & Infectious Diseases.

[8]  H. Pu,et al.  circMTDH.4/miR‐630/AEG‐1 axis participates in the regulation of proliferation, migration, invasion, chemoresistance, and radioresistance of NSCLC , 2019, Molecular carcinogenesis.

[9]  M. Brandi,et al.  Human Circulating miRNAs Real-time qRT-PCR-based Analysis: An Overview of Endogenous Reference Genes Used for Data Normalization , 2019, International journal of molecular sciences.

[10]  Jinha Hwang,et al.  MicroRNA Expression Profiles in Gastric Carcinogenesis , 2018, Scientific Reports.

[11]  A. Mishra,et al.  Emerging role of circulating microRNA in the diagnosis of human infectious diseases , 2018, Journal of cellular physiology.

[12]  M. Marinho,et al.  Characterization of the microtranscriptome of macrophages infected with virulent, attenuated and saprophyte strains of Leptospira spp. , 2018, PLoS neglected tropical diseases.

[13]  Mariko Matsui,et al.  Leptospirosis Pathophysiology: Into the Storm of Cytokines , 2018, Front. Cell. Infect. Microbiol..

[14]  V. Sitprija,et al.  Thai-Lepto-on-admission probability (THAI-LEPTO) score as an early tool for initial diagnosis of leptospirosis: Result from Thai-Lepto AKI study group , 2018, PLoS neglected tropical diseases.

[15]  R. Rettig,et al.  Identification of miR-16 as an endogenous reference gene for the normalization of urinary exosomal miRNA expression data from CKD patients , 2017, PloS one.

[16]  Ming O. Li,et al.  Regulation of the Immune Response by TGF-β: From Conception to Autoimmunity and Infection. , 2017, Cold Spring Harbor perspectives in biology.

[17]  Aaron Saguil,et al.  Evaluation of Jaundice in Adults. , 2017, American family physician.

[18]  Damian Szklarczyk,et al.  The STRING database in 2017: quality-controlled protein–protein association networks, made broadly accessible , 2016, Nucleic Acids Res..

[19]  Zhanguo Zhang,et al.  MicroRNA-630 suppresses tumor metastasis through the TGF-β- miR-630-Slug signaling pathway and correlates inversely with poor prognosis in hepatocellular carcinoma , 2016, Oncotarget.

[20]  R. Greil,et al.  miR-16-5p Is a Stably-Expressed Housekeeping MicroRNA in Breast Cancer Tissues from Primary Tumors and from Metastatic Sites , 2016, International journal of molecular sciences.

[21]  D. Bartel,et al.  Predicting effective microRNA target sites in mammalian mRNAs , 2015, eLife.

[22]  Hui Liu,et al.  Identification and validation of reference genes for the detection of serum microRNAs by reverse transcription-quantitative polymerase chain reaction in patients with bladder cancer. , 2015, Molecular medicine reports.

[23]  P. Newton,et al.  A Systematic Review of the Mortality from Untreated Leptospirosis , 2015, PLoS neglected tropical diseases.

[24]  Davide Heller,et al.  STRING v10: protein–protein interaction networks, integrated over the tree of life , 2014, Nucleic Acids Res..

[25]  Qingchuan Zhao,et al.  Increased MicroRNA-630 Expression in Gastric Cancer Is Associated with Poor Overall Survival , 2014, PloS one.

[26]  A. Rishi,et al.  Upregulation of miR-150* and miR-630 Induces Apoptosis in Pancreatic Cancer Cells by Targeting IGF-1R , 2013, PloS one.

[27]  E. Harhaj,et al.  RING Finger Protein 11 Targets TBK1/IKKi Kinases to Inhibit Antiviral Signaling , 2013, PloS one.

[28]  Xiaobing Fu,et al.  A Facile and Specific Assay for Quantifying MicroRNA by an Optimized RT-qPCR Approach , 2012, PloS one.

[29]  YOUHAI H. Chen,et al.  Ras family of small GTPases in immunity and inflammation. , 2012, Current opinion in pharmacology.

[30]  Guangchuang Yu,et al.  clusterProfiler: an R package for comparing biological themes among gene clusters. , 2012, Omics : a journal of integrative biology.

[31]  Brad T. Sherman,et al.  DAVID-WS: a stateful web service to facilitate gene/protein list analysis , 2012, Bioinform..

[32]  D. Bartel.  MicroRNAs: Target Recognition and Regulatory Functions , 2009, Cell.

[33]  P. Levett.  Leptospirosis , 2001, Clinical Microbiology Reviews.

[34]  D. Haake,et al.  Leptospirosis in humans. , 2015, Current topics in microbiology and immunology.

[35]  S. Knudsen,et al.  Evaluation of microRNA expression profiles that may predict recurrence of localized stage I non-small cell lung cancer after surgical resection. , 2010, Cancer research.