in vitro

Experimental validation of the network model:

The potential control points and effect modifiers identified through in silico approaches represent potential drug targets. We will experimentally manipulate these molecules (over-expression and knock-down) to validate our prediction model. We will also perform pair-wise perturbations for potential control points and effect modifiers to determine whether the pairs interact, in the sense that they modulate each other’s effect on the cellular phenotypes.

Model adjustment and validation for transformed HMECs:

We will introduce single or combined genetic alterations present in the cancer cell lines in our model and predict the steady states of the PI3K/MAPK network and the associated phenotypes. To validate these predictions, we will generate a panel of cell lines derived from HMECs carrying the identical genetic alterations including PI3K mutation, loss of
PTEN expression and oncogenic activation of RAS proteins,
alone or combinations thereof.