MiRNA182 regulates percentage of myeloid and erythroid cells in chronic myeloid leukemia
Authors: Cecil Ross
Sasikala P Sachithanandan
Deepak Arya
Shang Li
Dasaradhi Palakodeti
Sudhir Krishna
Journal: Cell death & disease (2017)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: lineage control programs; haematological malignancies; tyrosine kinase inhibitor resistance; chronic myeloid leukemia; microRNAs
Affiliations: Cellular Organization and Signalling Group, National Centre for Biological Sciences, Tata Institute of Fundamental Research, Bangalore, India., Manipal University, Manipal, India., Department of Medicine, St Johns Medical College and Hospitals, Bangalore, India., Stem Cells and Regeneration Group, Institute for Stem Cell Biology and Regenerative Medicine, Bangalore, India., Duke-NUS Graduate Medical School, Singapore.

Abstract:  The deregulation of lineage control programs is often associated with the progression of haematological malignancies. The molecular regulators of lineage choices in the context of tyrosine kinase inhibitor (TKI) resistance remain poorly understood in chronic myeloid leukemia (CML). To find a potential molecular regulator contributing to lineage distribution and TKI resistance, we undertook an RNA-sequencing approach for identifying microRNAs (miRNAs). Following an unbiased screen, elevated miRNA182-5p levels were detected in Bcr-Abl-inhibited K562 cells (CML blast crisis cell line) and in a panel of CML patients. Earlier, miRNA182-5p upregulation was reported in several solid tumours and haematological malignancies. We undertook a strategy involving transient modulation and CRISPR/Cas9 (clustered regularly interspersed short palindromic repeats)-mediated knockout of the MIR182 locus in CML cells. The lineage contribution was assessed by methylcellulose colony formation assay. The transient modulation of miRNA182-5p revealed a biased phenotype. Strikingly, Δ182 cells (homozygous deletion of MIR182 locus) produced a marked shift in lineage distribution. The phenotype was rescued by ectopic expression of miRNA182-5p in Δ182 cells. A bioinformatic analysis and Hes1 modulation data suggested that Hes1 could be a putative target of miRNA182-5p. A reciprocal relationship between miRNA182-5p and Hes1 was seen in the context of TK inhibition. In conclusion, we reveal a key role for miRNA182-5p in restricting the myeloid development of leukemic cells. We propose that the Δ182 cell line will be valuable in designing experiments for next-generation pharmacological interventions. 

Retracted: No

PubPeer comments:
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 Leuconostoc Miyukkimchii, why donu2019t you compare spots 51, 61, 62 and 71 to appreciate that it is not a case of background adjusted use of the same gel as you have accused. If you have not done 2-D gels then please do not discredit the experiments of others. You should apologize to the authors. , 
 The images are highly similar but there are key features that differ between the two. However, the first panel does have a numbering issue and the number 46 appears twice. , 
 Thanks for agreeing that the image differ between the to. The typgraphical error is deeply regretted. However, it wil not influence the outcome of the study. D. PARMAR , 
 ![file] Could the author post original blots. , 
 #5 ![file] It is a very impressive explanation given by Peperomia Verruculosa. It clearly shows stamping of selected regions of one blot to another. I also explored some some more regions of similarity highlighted in green circles or boxes. Even the background of both blots at lower left corner as very similar except its contrast, which is not possible even same samples run in 2D gel electrophoresis. This is clear cut case of fabrication of images using image editing tools. , 
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