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Updated: Jul 31, 2025
Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays
Published on: October 23, 2019
A customized mass array panel for BCR::ABL1 tyrosine kinase domain mutation screening in chronic myeloid leukemia
Nittaya Limsuwanachot1, Budsaba Rerkamnuaychoke1, Pimjai Niparuck2
1Department of Pathology, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, Thailand.
Introduction:
The therapeutic strategy and management of chronic myeloid leukemia (CML) have rapidly improved with the discovery of effective tyrosine kinase inhibitors (TKIs) to target BCR::ABL1 oncoprotein. However, nearly 30% of patients develop TKI resistance due to acquired mutations on the tyrosine kinase domain (TKD) of BCR::ABL1.
Methods:
We customized a mass array panel initially intended to detect and monitor the mutational burden of hotspot BCR::ABL1 TKD mutations accumulated in our database, including key mutations recently recommended by European LeukemiaNet. Additionally, we extended the feasibility of using the assay panel for the molecular classification of myeloproliferative neoplasms (MPNs) by incorporating primer sets specific for analyzing JAK2 V617F, MPL 515 K/L, and CALR types 1 and 2.
Results:
We found that the developed mass array panel was superior for detecting and monitoring clinically significant BCR::ABL1 TKD mutations, especially in cases with low mutational burden and harboring compound/polyclonal mutations, compared with direct sequencing. Moreover, our customized mass array panel detected common genetic alterations in MPNs, and the findings were consistent with those of other comparable assays available in our laboratory.
Conclusions:
Our customized mass array panel was practicably used as a routine robust assay for screening and monitoring BCR::ABL1 TKD mutations in patients with CML undergoing TKI treatment and feasible for analyzing common genetic mutations in MPNs.
Insights
A new mass array panel effectively detects BCR::ABL1 tyrosine kinase domain mutations in chronic myeloid leukemia (CML) patients, outperforming direct sequencing. It also aids in classifying myeloproliferative neoplasms (MPNs).
Area of Science:
- Hematology
- Molecular Diagnostics
- Oncology
Background:
- Tyrosine kinase inhibitors (TKIs) have transformed chronic myeloid leukemia (CML) treatment.
- BCR::ABL1 tyrosine kinase domain (TKD) mutations cause TKI resistance in nearly 30% of CML patients.
- Accurate detection of these mutations is crucial for effective CML management and understanding myeloproliferative neoplasms (MPNs).
Purpose of the Study:
- To develop and validate a customized mass array panel for detecting BCR::ABL1 TKD mutations.
- To assess the panel's utility in monitoring mutational burden and identifying resistance mutations in CML.
- To evaluate the panel's feasibility for molecular classification of MPNs, including JAK2, MPL, and CALR mutations.
Main Methods:
- Customization of a mass array panel to target hotspot BCR::ABL1 TKD mutations.
- Inclusion of primer sets for common MPN-associated mutations (JAK2 V617F, MPL 515 K/L, CALR types 1 and 2).
- Comparison of the mass array panel's performance against direct sequencing for BCR::ABL1 mutation detection.
Main Results:
- The developed mass array panel demonstrated superior sensitivity in detecting and monitoring clinically significant BCR::ABL1 TKD mutations, especially those with low mutational burden or compound/polyclonal patterns.
- The panel successfully identified common genetic alterations in MPNs, with results consistent with other laboratory assays.
- The assay proved effective in detecting mutations that confer TKI resistance in CML.
Conclusions:
- The customized mass array panel serves as a robust and practical routine assay for screening and monitoring BCR::ABL1 TKD mutations in CML patients undergoing TKI therapy.
- The panel is also feasible for analyzing common genetic mutations in MPNs, supporting molecular classification.
- This assay enhances the molecular monitoring capabilities for both CML and MPNs.
