title: MiRNA182 regulates percentage of myeloid and erythroid cells in chronic myeloid leukemia.
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.
authors.affiliation: Cellular Organization and Signalling Group, National Centre for Biological Sciences, Tata Institute of Fundamental Research, Bangalore, India.|Manipal University, Manipal, India.
authors.forename: Deepak
authors.initials: D
authors.lastname: Arya
authors.affiliation: Cellular Organization and Signalling Group, National Centre for Biological Sciences, Tata Institute of Fundamental Research, Bangalore, India.
authors.forename: Sasikala P
authors.initials: SP
authors.lastname: Sachithanandan
authors.affiliation: Department of Medicine, St Johns Medical College and Hospitals, Bangalore, India.
authors.forename: Cecil
authors.initials: C
authors.lastname: Ross
authors.affiliation: Stem Cells and Regeneration Group, Institute for Stem Cell Biology and Regenerative Medicine, Bangalore, India.
authors.forename: Dasaradhi
authors.initials: D
authors.lastname: Palakodeti
authors.affiliation: Duke-NUS Graduate Medical School, Singapore.
authors.forename: Shang
authors.initials: S
authors.lastname: Li
authors.affiliation: Cellular Organization and Signalling Group, National Centre for Biological Sciences, Tata Institute of Fundamental Research, Bangalore, India.
authors.forename: Sudhir
authors.initials: S
authors.lastname: Krishna
mesh_terms: D019070:Cell Lineage / Q000235:genetics; D049109:Cell Proliferation / Q000235:genetics*; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D041905:Erythroid Cells / Q000378:metabolism / Q000473:pathology; D016044:Fusion Proteins, bcr-abl / Q000235:genetics; D015973:Gene Expression Regulation, Leukemic / Q000187:drug effects; D006801:Humans; D020014:K562 Cells; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics* / Q000473:pathology; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D022423:Myeloid Cells / Q000378:metabolism / Q000473:pathology; D047428:Protein Kinase Inhibitors / Q000627:therapeutic use; D000072056:Transcription Factor HES-1 / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D035683:MicroRNAs; C538984:Mirn182 microRNA, human; D047428:Protein Kinase Inhibitors; D000072056:Transcription Factor HES-1; C078493:HES1 protein, human; D016044:Fusion Proteins, bcr-abl