Related Experiment Video
Updated: Mar 10, 2026
Author Spotlight: Integrating BRET-Based Assays and Rare Mutation Analysis to Decipher RAF Kinase Regulation in Live Cells
Published on: March 1, 2024
Improved FRET Biosensor for the Measurement of BCR-ABL Activity in Chronic Myeloid Leukemia Cells
Mika Horiguchi1, Mari Fujioka, Takeshi Kondo
1Department of Cell Physiology, Hokkaido University Graduate School of Medicine.
Abstract:
Although the co-development of companion diagnostics with molecular targeted drugs is desirable, truly efficient diagnostics are limited to diseases in which chromosomal translocations or overt mutations are clearly correlated with drug efficacy. Moreover, even for such diseases, few methods are available to predict whether drug administration is effective for each individual patient whose disease is expected to respond to the drug(s). We have previously developed a biosensor based on the principle of Förster resonance energy transfer to measure the activity of the tyrosine kinase BCR-ABL and its response to drug treatment in patient-derived chronic myeloid leukemia cells. The biosensor harbors CrkL, one of the major substrates of BCR-ABL, and is therefore named Pickles after phosphorylation indicator of CrkL en substrate. The efficacy of this technique as a clinical test has been demonstrated, but the number of cells available for analysis is limited in a case-dependent manner, owing to the cleavage of the biosensor in patient-derived leukemia cells. Here, we describe an improved biosensor with an amino acid substitution and a nuclear export signal being introduced. Of the two predicted cleavage positions in CrkL, the mutations inhibited one cleavage completely and the other cleavage partially, thus collectively increasing the number of cells available for drug evaluation. This improved version of the biosensor holds promise in the future development of companion diagnostics to predict responses to tyrosine kinase inhibitors in patients with chronic myeloid leukemia.
Insights
Researchers improved a biosensor for chronic myeloid leukemia (CML) to better predict tyrosine kinase inhibitor response. This enhanced diagnostic tool increases available cells for drug evaluation, aiding personalized medicine in CML treatment.
Area of Science:
- Biotechnology and Biomedical Engineering
- Oncology and Hematology
- Molecular Diagnostics
Background:
- Co-development of companion diagnostics with targeted therapies is crucial but limited by the lack of efficient predictive methods.
- Existing diagnostics for chronic myeloid leukemia (CML) struggle with limited cell availability for drug response evaluation.
- Previous Förster resonance energy transfer (FRET)-based biosensor (Pickles) showed promise but suffered from biosensor cleavage in patient cells.
Purpose of the Study:
- To develop an improved biosensor for enhanced drug evaluation in chronic myeloid leukemia (CML) patient cells.
- To increase the number of available cells for analyzing tyrosine kinase inhibitor (TKI) response.
- To advance the development of companion diagnostics for personalized CML therapy.
Main Methods:
- Engineered a Förster resonance energy transfer (FRET)-based biosensor incorporating CrkL (CrkL phosphorylation indicator en substrate).
- Introduced an amino acid substitution and a nuclear export signal into the biosensor to inhibit cleavage.
- Evaluated the improved biosensor's performance in patient-derived CML cells for drug response assessment.
Main Results:
- The improved biosensor demonstrated significantly reduced cleavage at predicted sites within the CrkL substrate.
- Mutations effectively inhibited one cleavage site completely and another partially, increasing cell availability for analysis.
- The enhanced biosensor facilitates more robust evaluation of tyrosine kinase inhibitor efficacy in CML.
Conclusions:
- The modified biosensor overcomes limitations of previous versions by enhancing cell availability for drug response testing.
- This improved diagnostic tool holds significant potential for predicting patient response to tyrosine kinase inhibitors in CML.
- The study paves the way for more effective companion diagnostics in personalized cancer therapy.
More Related Videos
09:57Comprehensive Protocol to Sample and Process Bone Marrow for Measuring Measurable Residual Disease and Leukemic Stem Cells in Acute Myeloid Leukemia
Published on: March 5, 2018
08:27Detection of Human Leukocyte Antigen Biomarkers in Breast Cancer Utilizing Label-free Biosensor Technology
Published on: March 24, 2015