Aims
In collaboration with the Industrial Partners, cellular screening systems will be developed to detect apoptosis modulating properties of candidate drugs. The following approaches will form the basis of the screening systems.
I.Enhancement/inhibition of Fas receptor-mediated apoptosis by targeting anti-apoptotic signaling. To evaluate whether the drugs of interest could activate/inhibit anti-apoptotic signaling, we will test facilitation of Fas receptor responsiveness in the same way as presented in Fig. 1 in background
II.Inhibition/activation of HSP70 synthesis (enhancement/inhibition of apoptosis). The aim is to evaluate whether a drug could induce or inhibit a heat shock stress response and, thereby act as an apoptotic modulator. For the detection of a stress response, we will use standard methods used in our laboratories for detection of transcriptional activation of stress genes (activation of heat shock factor 1, measurement of HSP70 protein levels) and activation of heat shock signaling (activation of HSFs).
Fig. 2. An example of a screening approach. Here a stable HeLa cell line has been generated, which expresses green fluorescent protein (GFP) under the control of the heat shock promoter element. When a cell is stressed the expression of GFP is activated and can be easily measured by a fluorescence scanner. The inhibition of GFP expression is a measure of the anti-Hsp70 potential of a given drug. The basic principle of this idea is presented in Fig. 2A. The same construct can also be used for screening drugs that are activating the stress response (Fig. 2B).