Dysregulation of VDR-associated lncRNAs in patients with multiple sclerosis
Dysregulation of VDR-associated lncRNAs in patients with multiple sclerosis
Shahrokh Janamiri,1Somayeh Farahmand,2,*Mehdi Dadmehr,3
1. Department of Biology, Payame Noor University (PNU) 2. Department of Biology, Payame Noor University (PNU) 3. Department of Biology, Payame Noor University (PNU)
Introduction: Long non-coding RNAs (lncRNAs) have been shown to influence pathoetiology of multiple sclerosis (MS). We aimed at identification of expression levels of three vitamin D receptor-associated lncRNAs, namely SNHG6, SNHG16 and LINC00346 in the circulation of MS patients compared with healthy subjects. Expression of SNHG6 was significantly lower in MS patients compared with controls (expression ratio (ER) (95% CI)= 0.39 (0.22-0.69), adjusted P value=0.0015) and in female patients compared with female controls (ER (95% CI)= 0.28 (0.13-0.59), adjusted P value=0.0001). Expression of SNHG16 was also lower in total MS patients compared with total controls (ER (95% CI)= 0.24 (0.1-0.57), adjusted P value=0.0001). Expression of LINC00346 was lower in total patients compared with controls (ER (95% CI)= 0.03 (0.009-0.09), adjusted P value<0.0001).
Methods: Four mL of whole peripheral blood was acquired via venipuncture from MS patients and healthy subjects in EDTA tubes. Hybrid-RTM blood RNA extraction kit (GeneAll Biotechnology Co Ltd., South Koera) was used for extraction of total RNA. After verification of appropriate quality and concentration of RNA, cDNA was synthesized using High-Capacity cDNA Reverse Transcription kit (Applied Biosystems). Expression of SNHG6, SNHG16 and LINC00346 were quantified in the rotor gene 6000 Corbett Real-Time PCR System by using SYBR® Premix Ex TaqTM (TaKaRa, Japan). B2M gene was used as normalizer.The Statistical Package for the Social Sciences (SPSS) v.18.0 (SPSS Inc., Chicago, IL) was used for statistical assessments. GraphPad Prism version 9.0 for Windows (GraphPad Software, La Jolla California, USA) was used for depiction of graphs.
Results: There was no significant difference in expressions of SNHG6, SNHG16 and LINC00346 between male and female patients. There was no significant correlation between expressions of SNHG6, SNHG16 and LINC00346 lncRNAs and age, disease duration, age at onset or EDSS. LINC00346 had the AUC values of 0.84, 0.82 and 0.94 in differentiation of total MS patients from total controls, female patients from female controls and male patients from male controls, respectively. SNHG6 could separate female patients from female controls with AUC of 0.79. Finally, SNHG16 could separate male patients from male controls with AUC of 0.73.
Conclusion: these lncRNAs might be proposed as putative peripheral markers for MS and potential contributors in the pathogenesis of MS.