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Feb 15, 2010 (Vol. 30, No. 4)

Refined Tools Aid in Pathway Analysis

Scientists Explore Range of Options to Remove Roadblocks Along the New Product Development Highway

  • Effects of HSP90 Inhibition

    Click Image To Enlarge +
    A merged multichannel image of lung cancer cells stained for DNA and for cell-cycle phase markers (Exelixis)

    Susan Lyman, Ph.D., research scientist at Exelixis, presented a cell-cycle analysis method that she and her colleagues developed. “This new method allows us to easily profile cell-cycle perturbations in a large number of samples, with improved throughput and greater information content versus classical methods of cell-cycle analysis like FACS.”

    Dr. Lyman noted that cell-cycle analysis has traditionally been carried out by fluorescence activated cell sorting (FACS). The new method, in contrast, uses image-based assessment of fluorescently labeled cells, combining a measurement of DNA content with imaging of several phase-specific readouts.

    “The end result is a novel and robust high-throughput Cellomics-based cell-cycle assay that reports the phase status of a cell as well as its DNA content.” Dr. Lyman and her colleagues applied this technique to examine the cell-cycle perturbations caused by inhibition of HSP90, a molecular chaperone that enhances the stability of client proteins. 

    “We used the high-content cell-cycle method to analyze the cell-cycle effects of several different small molecule inhibitors of HSP90 in a large panel of cancer cell lines and were amazed at how distinct HSP90 inhibitors yielded nearly identical cell-cycle phenotypes. It’s been a very useful tool for profiling small molecules,” Dr. Lyman insisted.

    “In developing this cell-cycle analysis method, we tried to combine the best aspects of FACS and the best aspects of high-content imaging to generate high-quality cell-profiling data for our internal preclinical studies. Essentially, we have created a new application of some existing technologies to produce an efficient and robust means of assessing cell-cycle perturbations to help drive our preclinical development.”

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