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Updated: May 30, 2025
Pre-clinical Evaluation of Tyrosine Kinase Inhibitors for Treatment of Acute Leukemia
Published on: September 18, 2013
Sensitivity to Tyrosine Kinase Inhibitors in a Human Philadelphia Chromosome-Positive (Ph+) Leukemia Model With the
Thao T Nguyen1, Minori Tamai1, Daisuke Harama1
1Department of Pediatrics, University of Yamanashi, Chuo, JPN.
Abstract:
The T315I-inclusive compound mutation, the multiple mutations including the T315I mutation on the same BCR::ABL1 gene, confers resistance to diverse tyrosine kinase inhibitors (TKIs). Development of the F311I/T315I compound mutation has been reported in chronic myeloid leukemia patients who sequentially showed clinical resistance to imatinib and dasatinib. The establishment of a human leukemia model with the T315I-inclusive compound mutation remains an experimental challenge. Here, we introduced the F311I/T315I compound mutation into the intrinsic BCR::ABL1 gene of a human TKI-sensitive Philadelphia chromosome-positive leukemia cell line via homologous recombination using the CRISPR/Cas9 system and obtained three types of sublines: the F311I mutation alone, the T315I mutation alone, and the F311I/T315I compound mutation. The F311I subline was sensitive to dasatinib but moderately resistant to imatinib and nilotinib, while the T315I subline and the F311I/T315I subline were highly resistant to these TKIs. Notably, the T315I subline and the F311I/T315I subline were sensitive to therapeutic concentrations of ponatinib, although more resistant than the F311I subline. Moreover, the T315 subline and the F311I/T315 subline were sensitive to asciminib at therapeutic concentration, as was the F311I subline. This is the first human leukemia model in which the impact of the T315I-inclusive compound mutation on TKI sensitivity was directly confirmed.
Insights
The F311I/T315I compound mutation in the BCR::ABL1 gene causes resistance to tyrosine kinase inhibitors (TKIs). This study created a human leukemia model to confirm this resistance and test new drug sensitivities.
Area of Science:
- Hematology
- Molecular Biology
- Genetics
Background:
- The T315I mutation in the BCR::ABL1 gene confers resistance to tyrosine kinase inhibitors (TKIs), a common challenge in treating chronic myeloid leukemia (CML).
- Compound mutations, such as F311I/T315I, further complicate TKI resistance, necessitating novel therapeutic strategies.
- Establishing reliable human leukemia models for studying these complex mutations is crucial for preclinical research.
Purpose of the Study:
- To establish a human leukemia model harboring the F311I/T315I compound mutation in the BCR::ABL1 gene.
- To investigate the impact of the F311I/T315I compound mutation on sensitivity to various TKIs.
- To provide a platform for evaluating novel therapeutic agents against TKI-resistant leukemia.
Main Methods:
- Utilized CRISPR/Cas9 gene editing and homologous recombination to introduce specific mutations (F311I, T315I, and F311I/T315I) into a TKI-sensitive human leukemia cell line.
- Generated distinct sublines representing each mutation for comparative analysis.
- Assessed the sensitivity of these sublines to imatinib, dasatinib, nilotinib, ponatinib, and asciminib at therapeutic concentrations.
Main Results:
- The F311I mutation alone conferred moderate resistance to imatinib and nilotinib but retained sensitivity to dasatinib.
- Both the T315I single mutation and the F311I/T315I compound mutation resulted in high resistance to imatinib, dasatinib, and nilotinib.
- The T315I and F311I/T315I sublines demonstrated sensitivity to ponatinib and asciminib at therapeutic doses, though with varying degrees of resistance compared to the F311I subline.
Conclusions:
- This study successfully established the first human leukemia model to directly demonstrate the impact of the T315I-inclusive compound mutation on TKI resistance.
- The model confirms the significant resistance conferred by the F311I/T315I mutation to several first- and second-generation TKIs.
- The findings highlight the potential of ponatinib and asciminib as effective therapeutic options for leukemia patients with T315I-inclusive compound mutations.
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