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Development and application of fluorescence bioimaging technique

Clinical application of FRET biosensors

Fluorescence bioimaging can detect protein-protein interaction or protein conformational change with high sensitivity and quantitativity in living cells. Our laboratory utilizes the merit and applies fluorescent protein to clinical test.
Chronic myeloid leukemia (CML) is a hematopoietic malignancy characterized by the presence of the Philadelphia chromosome (Ph). The Ph results from a reciprocal translocation, leading to the formation of a novel fusion protein, BCR-ABL.
Recently, tyrosine kinase inhibitors of BCR-ABL were clinically approved. However, the efficacy of the drugs could not be judged before treatment, but be speculated by clinical courses after drug administration. Clinical testing to predict drug efficacy before treatment is therefore desired, which promise patients effective and secure therapy.
We develop a FRET biosensor, Pickles (Phosphorylation indicator of CrkL en substrate, Figure1), that utilizes CrkL, the most characteristic substrate of BCR-ABL. This biosensor successfully monitored the drug response in CML cells from the patients (Figure 2).
Appling this technique to CML patients enables us to predict the most effective drug against each patient before treatment. In addition, because cells are alive during this assay, drug resistant cells can be collected based on FRET efficiency and subjected to further analyses. This may help us to elucidate the mechanism of drug resistance in future.

bio_censor01

 

 

  

bio_censor02

 

【Related papers】

  • A novel FRET-based biosensor for the measurement of BCR-ABL activity and its response to drugs in living cells. T. Mizutani, T. Kondo, S. Darmanin, M. Tsuda, S. Tanaka, M. Tobiume, M. Asaka, & Y. Ohba*. Clin. Cancer Res. 16(15): 3964-3975 (2010)

  • Commentary on this paper: Clin. Cancer Res. 16(15): 3822-3824 (2010) 

 

 

【Related information】

 

 

Generation of organelle marker library

We have generated the organelle marker library to visualize various organelles, including nuclei, plasma membrane, actin, microtubules (tubulin), focal contacts, mitochondria, early endosomes, late endosomes, recycling endosomes, the Golgi, and ER. Each organelle marker fused with a series of fluorescent proteins: e.g. Sirius, SECFP, EGFP, Venus, tdTomato, TFP650, and Keima.

 

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