RT Journal Article
T1 A yeast-based in vivo bioassay to screen for class I phosphatidylinositol 3-kinase specific inhibitors.
A1 Fernández-Acero Bascones, Teresa
A1 Rodríguez Escudero, María Isabel
A1 Vicente, Francisca
A1 Monteiro, Maria Cândida
A1 Tormo, José R.
A1 Cantizani, Juan
A1 Molina, María
A1 Cid, Víctor J.
AB The phosphatidylinositol 3-kinase (PI3K) pathway couples receptor-mediated signaling to essential cellular functions by generating the lipid second messenger phosphatidylinositol-3,4,5-trisphosphate. This pathway is implicated in multiple aspects of oncogenesis. A low-cost bioassay that readily measures PI3K inhibition in vivo would serve as a valuable tool for research in this field. Using heterologous expression, we have previously reconstituted the PI3K pathway in the model organism Saccharomyces cerevisiae. On the basis of the fact that the overproduction of PI3K is toxic in yeast, we tested the ability of commercial PI3K inhibitors to rescue cell growth. All compounds tested counteracted the PI3K-induced toxicity. Among them, 15e and PI-103 were the most active. Strategies to raise the intracellular drug concentration, specifically the use of 0.003% sodium dodecyl sulfate and the elimination of the Snq2 detoxification pump, optimized the bioassay by enhancing its sensitivity. The humanized yeast-based assay was then tested on a pilot scale for high-throughput screening (HTS) purposes using a collection of natural products of microbial origin. From 9600 extracts tested, 0.6% led to a recovery of yeast growth reproducibly, selectively, and in a dose-dependent manner. Cumulatively, we show that the developed PI3K inhibition bioassay is robust and applicable to large-scale HTS.
SN 1552-454X
YR 2012
FD 2012-06-15
LK https://hdl.handle.net/20.500.14352/43862
UL https://hdl.handle.net/20.500.14352/43862
LA eng
NO Fernández-Acero Bascones, T., Rodríguez Escudero, M. I., Vicente, F. et al. «A Yeast-Based In Vivo Bioassay to Screen for Class I Phosphatidylinositol 3-Kinase Specific Inhibitors». SLAS Discovery, vol. 17, n.o 8, septiembre de 2012, pp. 1018-29. DOI.org (Crossref), https://doi.org/10.1177/1087057112450051.
NO Ministerio de Ciencia, Innovación y Universidades (España)
NO Universidad Complutense de Madrid/Banco de Santander
NO Fundación MEDINA, a public-private partnership of Merck Sharp & Dohme de España S.A./Universidad de Granada/Junta de Andalucía. T.F-A
NO Ministerio de Educación, Formación Profesional y Deportes (España)
DS Docta Complutense
RD 5 ago 2024