A novel microRNA boosts hyper-β-oxidation of fatty acids in liver by impeding CEP350-mediated sequestration of PPARα and thus restricts chronic hepatitis C
Authors: Joyeeta Chakraborty
Susree Roy
Raghunath Chatterjee
Amit Ghosh
Soma Banerjee
Suvendra Nath Bhattacharyya
Avijit Goswami
Sayantani Bhowmik
Sakshi Dokania
Simanti Datta
Abhijit Chowdhury
Suchandrima Ghosh
Priyanka Kumari
Journal: RNA Biology (2020)
Discipline: BIOLOGY & BIOCHEMISTRY
Keywords: HCV; Lipid metabolism; hepatitis C virus; microrna; novel microRNA.
Affiliations: Centre for Liver Research, School of Digestive and Liver Diseases,  Institute of Post Graduate Medical Education and Research , Kolkata, India, Human Genetics Unit,  Indian Statistical Institute , Kolkata, Human Genetics Unit, India, Department of Molecular Genetics,  Indian Institute of Chemical Biology , Kolkata, India, Department of Hepatology,  School of Digestive and Liver Diseases, Institute of Post Graduate Medical Education and Research , Kolkata, India

Abstract:  Imbalance in lipid metabolism induces steatosis in liver during Chronic hepatitis C (CHC). Contribution of microRNAs in regulating lipid homoeostasis and liver disease progression is well established using small RNA-transcriptome data. Owing to the complexity in the development of liver diseases, the existence and functional importance of yet undiscovered regulatory miRNAs in disease pathogenesis was explored in this study using the unmapped sequences of the transcriptome data of HCV-HCC liver tissues following miRDeep2.pl pipeline. MicroRNA-c12 derived from the first intron of LGR5 of chromosome 12 was identified as one of the miRNA like sequences retrieved in this analysis that showed human specific origin. Northern blot hybridization has proved its existence in the hepatic cell line. Enrichment of premiR-c12 in dicer-deficient cells and miR-c12 in Ago2-RISC complex clearly suggested that it followed canonical miRNA biogenesis pathway and accomplished its regulatory function. Expression of this miRNA was quite low in CHC tissues than normal liver implying HCV-proteins might be regulating its biogenesis. Promoter scanning and ChIP analysis further revealed that under expression of p53 and hyper-methylation of STAT3 binding site upon HCV infection restricted its expression in CHC tissues. Centrosomal protein 350 (CEP350), which sequestered PPARα, was identified as one of the targets of miR-c12 using Miranda and validated by luciferase assay/western blot analysis. Furthermore, reduced triglyceride accumulation and enhanced PPARα mediated transcription of β-oxidation genes upon restoration of miR-c12 in liver cells suggested its role in lipid catabolism. Thus this study is reporting miR-c12 for the first time and showed its' protective role during chronic HCV infection. 

Retracted: No

PubPeer comments:
 ![file] , 
 Checked this image from the paper after changing the brightness and contrast. I do not think there is any duplication. ![file] , 
 I do agree with #Dracophyllum Balansae. There is a duplication between lane 3 and 4 of histone H3 panel. I have also found an splicing mark between lane 1 and 2.![file] , 
 We are investigating this issue and looking into the original blot carefully. If we find any mistake by the main author of this manuscript, we will contact the journal editor as soon as possible. The experiment was performed multiple times. We will submit all the data. , 
 #2 I disagree. The duplication is obvious here, and the splice confirms that. When an image at low resolution is duplicated, compression artifacts occur, and little differences at pixel level are recognizable, although the image is the same. Here similarities of band shape and internal features within the bands are too high to be other than a duplication. , 
 We have contacted the Editor-In-Chief with corrected figure. The correction is in progress. , 
 #6 Could the authors please post the uncropped scans? It is difficult to understand how this could occur by chance. , 
 Correction Published online: 25 Aug 2021  In Figure 4e, incorrect control panel (sic) (Histone H3) was used. Western blot for PPARu03b1 and Histone H3 were repeated and band intensity ratios were calculated. Figure 4e. ![file] In Figure 5c, the same (sic) image was used for premir-c12 and anti-sense CEP350 treatment panels. Anti-sense CEP350 has been replaced with correct picture. Figure 5c. ![file] The final conclusion of each figure remains same as presented in the article. Disappointing that the authors did not address the actual concerns in this correction. ,