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.

miR-182-5p Promotes Growth in Oral Squamous Cell Carcinoma by Inhibiting CAMK2N1

“…miR-182-5p is a highly conserved miRNA whose expression has been found to be increased in several malignant human cancers, such as glioblastoma, hepatocellular carcinoma, and breast and gastric cancer [14]. However, how miR-182-5p contributes to the proliferation of OSCC cells remains poorly characterized.…”

“…miRNAs are known to be an integral component of cancer development [17][18][19]. Several studies have highlighted the role of miR-182-5p in the development of different cancers [13,14]. In glioblastoma, miR-182-5p targets protein phosphatase 1 regulatory inhibitor subunit 1C [20].…”

CRISPR/Cas9 Age in Genome Editing and Leukemia Applications

“…KML hücre hattı K-562'de miR182-5p'nin, tirozin kinaz inhibitörlerine (TKI) karşı gelişen direnç üzerine etkisi araştırmak amacı ile CRISPR/Cas9 teknolojisi ile miR182 lokusu homozigot olarak inhibe edilerek; miR182-5p ekspresyonu azaltılmıştır. Böylelikle, myeloid hücre proliferasyonu engellenerek, hücre farklılaşması sağlanmış, eritrosit çoğalması indüklenmiş ve kemotedavi direnci kırılarak, TKI sensitizasyonu sağlanmıştır 102 . Akut monositik lösemi hücre hattı MV4-11 ile yapılan farklı bir çalışmada, lösemi hücre büyümesi üzerinde etkili miRNA'ları belirlemek için CRISPR/Cas9 kütüphanesi (lentiCRISPRv2 kütüphanesi) oluşturulmuştur.…”

Emerging Role of CRISPR/Cas9 Technology for MicroRNAs Editing in Cancer Research

“…There has been an interest in analyzing the role of miRNAs in hematopoiesis and related disease states (40). In a recent study, it was reported that myeloid differentiation in chronic myeloid leukemia cells (K562) is mediated by the loss of miRNA182-5p expression (41). In this study, Arya and colleagues used the CRISPR/Cas gene-editing approach to targeting miRNA182 locus and abrogate miRNA182-5p expression.…”