Abstract 1379: High throughput screening to identify molecules that alter VHL stability Academic Article uri icon

abstract

  • Abstract Von Hippel-Lindau (VHL) germ line and somatic mutations play determining roles in renal-cell carcinomas (RCC) as well as other diseases, including hemangioblastomas and pheochromocytomas. Although molecules upregulated as a consequence of VHL deficiency have been employed as drug targets in RCC cancer therapy, modulation of pVHL itself has not been explored as a therapeutic strategy in the treatment of VHL derived diseases. Approximately one third of VHL mutations are point mutations, which destabilize pVHL and decrease pVHL intracellular levels, despite in some cases possessing residual functionality. Therefore, interventions that restabilize mutated pVHL may restore physiological protein levels and function. We established a high-throughput screening system to identify small molecules that alter VHL stability. We fused an unstable W117A VHL mutant to a fluorescent protein Venus and transduced the VHL null 786-0 RCC cell line. The fluorescence and protein levels of W117A VHL-Venus were low in untreated cells, and increased significantly after treatment with the proteasome inhibitors MG132 or bortezomib. The fluorescence level served as a VHL protein stability readout for the screen. We then performed a screen on the Prestwick drug library and identified multiple candidates that stabilized VHL protein. Certain candidates have been shown to functionally regulate VHL stability as well as its downstream effecter HIF1a. Our work shows that the high-throughput screening system we established could efficiently identify small molecules that alter VHL stability and functionality, and facilitates exploring VHL as a drug target in the treatment of RCC as well as other VHL diseases. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1379. doi:10.1158/1538-7445.AM2011-1379

published proceedings

  • Cancer Research

author list (cited authors)

  • Ding, Z., Feng, Z., Fikes, N. A., Gao, M., Si, W., Sobieski, M. M., ... Jonasch, E.

citation count

  • 0

complete list of authors

  • Ding, Zhiyong||Feng, Zhehui||Fikes, Nelda A||Gao, Meng||Si, Wendy||Sobieski, Mary M||Stephan, Clifford C||Mills, Gordon B||Jonasch, Eric

publication date

  • January 2011