Abstract
Background: The t(12;21)(p13;q22), which fuses ETV6 and RUNX1 genes, is the most common genetic abnormality in children with B-cell precursor acute lymphoblastic leukaemia. The implication of the fusion protein in leukemogenesis seems to be clear. However, its role in the maintenance of the disease continues to be controversial. Methods: Generation of an in vitro ETV6/RUNX1 knock out model using the CRISPR/Cas9 gene editing system. Functional characterization by RNA sequencing, proliferation assays, apoptosis and pharmacologic studies, and generation of edited-cell xenograft model. Results: The expression of ETV6/RUNX1 fusion gene was completely eliminated, thus generating a powerful model on which to study the role of the fusion gene in leukemic cells. The loss of fusion gene expression led to the deregulation of biological processes affecting survival such as apoptosis resistance and cell proliferation capacity. Tumour cells showed higher levels of apoptosis, lower proliferation rate and a greater sensitivity to PI3K inhibitors in vitro along as a decrease in tumour growth in xenografts models after ETV6/RUNX1 fusion gene abrogation. Conclusions: ETV6/RUNX1 fusion protein seems to play an important role in the maintenance of the leukemic phenotype and could thus become a potential therapeutic target.
Highlights
The t(12;21)(p13;q22), which fuses ETV6 and RUNX1 genes, is the most common genetic abnormality in children with B-cell precursor acute lymphoblastic leukaemia.The implication of the fusion protein in leukemogenesis seems to be clear
E/R sequence was edited by CRISPR/Cas9 and evaluated by Sanger sequencing in REH cells (Figure S1)
KO1 clone carried an insertion of two cytosines at the end of ETV6 exon 5, near the Protospacer Adjacent Motif (PAM) sequence of the sgRNA G2, a frameshift mutation that generated a stop codon before finishing the exon
Summary
The t(12;21)(p13;q22), which fuses ETV6 and RUNX1 genes, is the most common genetic abnormality in children with B-cell precursor acute lymphoblastic leukaemia.The implication of the fusion protein in leukemogenesis seems to be clear. The gene fusion between the transcription factors ETV6 (TEL) and RUNX1 (AML1), generated by t(12;21)(p13;q22), is the most frequent chromosomal translocation in children with acute lymphoblastic leukaemia (ALL). This translocation fuses the 50 non-DNA binding region of the ETS family transcription factor ETV6 to almost the entire RUNX1 locus [1,2]. Patients carrying this translocation are associated with a good prognosis and excellent molecular response to treatment.
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