Thursday, November 21, 2024
11/21/2024
PROJECT SUMMARY Over the past 30 years, the field of protein kinase inhibitors development has rapidly expanded. As a result, several dozens of small-molecule kinase inhibitors have been approved by FDA, that led to improved outcomes in cancer treatment for thousands of patients. Moreover, kinase inhibition was recently extended towards non- oncology applications such as autoimmune and inflammatory diseases. The ultimate success of a drug development process relies on the availability of prompt, reliable, and cost- efficient analytical methodologies for potential inhibitor profiling. In the case of kinase inhibition, the screening is often performed against purified recombinant kinase domains in vitro. While this approach has led to successful identification of kinase inhibitors, it suffers from some major limitations that hamper drug development process. A critical limitation is the poor correlation between in vitro and cellular behavior of inhibitor candidates. This limitation is a reason for late-in-the-process failures of some drug candidates, that overall contribute to high cost of the drug development process. This limitation can be overcome via development and use of highly efficient cellular assays for inhibitor profiling. However, currently there are no efficient cellular assays for kinase inhibitor profiling in native environments. In the proposed work, we will design and develop a new cellular assay for profiling inhibitors of a model kinase, EGFR tyrosine kinase. The assay will enable screening inhibitor activity in native conditions, inside the living cells. To develop the cellular assay, we will accomplish three specific aims: (1) rational design, synthesis, and in vitro characterization of the new cell-permeable fluorescent sensors that selectively respond on low-nanomolar concentrations of EGFR; (2) development and characterization of intracellular delivery, localization, and EGFR binding of these new sensors; (3) development of a competitive cellular assay for inhibitors profiling and quantitative assessment of the inhibitor binding. Upon successful completion of this work, the new assay will not only provide a foundation for the high-throughput profiling of EGFR inhibitors, but also will establish general development principles applicable for other protein targets. In addition, this project will enhance research environment and infrastructure at Northern Illinois University. Thus, generations of undergraduate students will get valuable experience in design and synthesis of the new sensors, characterizing biomolecule/sensor interaction, working on developing in vitro and cellular assays. Eventually, participation in this project will prepare and empower the undergraduate students for future careers in biomedical field.
HHS’ Tracking Accountability in Government Grants System (TAGGS) website provides access to detailed descriptions of grants, loans, aggregated direct payments and other types of financial assistance awarded by the HHS Operating Divisions (OPDIVs) and the Office of the Secretary Staff Divisions (STAFFDIVs).
