Flow Cytometric Assay of Telomerase Activity

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DESCRIPTION (provided by applicant): The potential contribution of telomerase activity towards unregulated cell proliferation and tumor growth has yet to be determined, and currently used methods for the measurement of telomerase activity are too cumbersome to be employed in the clinic, or used to facilitate pharmacologic screening. There is currently an acute and urgent need for a rapid telomerase assay. The proposed highly innovative program should lead to the development of a non-radioactive, rapid, inexpensive, sensitive, and ultimately homogeneous and routine flow-cytometric assay for telomerase activity. At first, this novel assay will be bead-based, employ a PCR step to maintain sensitivity, and function without the need for analytical gels. The latter half of our research plan involves optimization of our bead-based technology to eliminate the PCR step. Our bead-based assay would establish the needed expertise to develop routine diagnostic and prognostic assays for the detection of telomerase activity associated with diseases that involve undesired cell proliferation. Furthermore, the establishment of expertise regarding the development of bead-based assays of polymerases could foster the development of analogous tests designed to detect the selective transcriptional activation of interesting genes with prognostic significance. We describe a robust and thorough research plan that describes how we are highly likely to succeed in our goal to develop a much needed assay for the routine detection of telomerase activity. The optimization of telomerase activity detection should thereafter lead to the development, in phase II, of solid phase multiplex analysis that would be used to screen thousands of compounds for their ability to inhibit telomerase activity. Such compounds might be helpful in augmenting currently used agents to selectively interfere with the ability of tumor cells to replicate indefinitely. Thus, upon completion of phase I, we will have developed telomerase assays suitable for routine clinical, and research use, and upon completion of phase II we will have an assay suitable for drug discovery use.