IN DUBIO PRO NATURA? A Philosophical Analysis of the Precautionary Principle in Environmental and Health Risk Governance

In this book, I apply a philosophical approach to study the precautionary principle in environmental (and health) risk decision-making. The principle says that unacceptable environmental and health risks should be anticipated, and they ought to be forestalled before the damage comes to fruition even if scientific understanding of the risks is inadequate. The study consists of introductory chapters, summary and seven original publications which aim at explicating the principle, critically analysing the debate on the principle, and constructing a basis for the well-founded use of the principle. Papers I-V present the main thesis of this research. In the two last papers, the discussion is widened to new directions. The starting question is how well the currently embraced precautionary principle stands up to critical philosophical scrutiny. The approach employed is analytical: mainly conceptual, argumentative and ethical. The study draws upon Anglo-American style philosophy on the one hand, and upon sources of law as well as concrete cases and decision-making practices at the European Union level and in its member countries on the other. The framework is environmental (and health) risk governance, including the related law and policy. The main thesis of this study is that the debate on the precautionary principle needs to be shifted from the question of whether the principle (or its weak or strong interpretation) is well-grounded in general to questions about the theoretical plausibility and ethical and socio-political justifiability of specific understandings of the principle. The real picture of the precautionary principle is more complex than that found (i.e. presumed) in much of the current academic, political and public debate surrounding it. While certain presumptions and interpretations of the principle are found to be sound, others are theoretically flawed or include serious practical problems. The analysis discloses conceptual and ethical presumptions and elementary understandings of the precautionary principle, critically assesses current practices invoked in the name of the precautionary principle and public participation, and seeks to build bridges between precaution, engagement and philosophical ethics. Hence, it is intended to provide a sound basis upon which subsequent academic scrutiny can build.

Targeting oncogenic signaling proteins : new insights using a FRET-based chemical biology approach

Cancer affects more than 20 million people each year and this rate is increasing globally. The Ras/MAPK-pathway is one of the best-studied cancer signaling pathways. Ras proteins are mutated in almost 20% of all human cancers and despite numerous efforts, no effective therapy that specifically targets Ras is available to date. It is now well established that Ras proteins laterally segregate on the plasma membrane into transient nanoscale signaling complexes called nanoclusters. These Ras nanoclusters are essential for the high-fidelity signal transmission. Disruption of nanoclustering leads to reduction in Ras activity and signaling, therefore targeting nanoclusters opens up important new therapeutic possibilities in cancer. This work describes three different studies exploring the idea of membrane protein nanoclusters as novel anti-cancer drug targets. It is focused on the design and implementation of a simple, cell-based Förster Resonance Energy Transfer (FRET)-biosensor screening platform to identify compounds that affect Ras membrane organization and nanoclustering. Chemical libraries from different sources were tested and a number of potential hit molecules were validated on full-length oncogenic proteins using a combination of imaging, biochemical and transformation assays. In the first study, a small chemical library was screened using H-ras derived FRET-biosensors. Surprisingly from this screen, commonly used protein synthesis inhibitors (PSIs) were found to specifically increase H-ras nanoclustering and downstream signalling in a H-ras dependent manner. Using a representative PSI, increase in H-ras activity was shown to induce cancer stem cell (CSC)-enriched mammosphere formation and tumor growth of breast cancer cells. Moreover, PSIs do not increase K-ras nanoclustering, making this screening approach suitable for identifying Ras isoform-specific inhibitors. In the second study, a nanoncluster-directed screen using both H- and K-ras derived FRET biosensors identified CSC inhibitor salinomycin to specifically inhibit K-ras nanocluster organization and downstream signaling. A K-ras nanoclusteringassociated gene signature was established that predicts the drug sensitivity of cancer cells to CSC inhibitors. Interestingly, almost 8% of patient tumor samples in the The Cancer Genome Atlas (TCGA) database had the above gene signature and were associated with a significantly higher mortality. From this mechanistic insight, an additional microbial metabolite screen on H- and K-ras biosensors identified ophiobolin A and conglobatin A to specifically affect K-ras nanoclustering and to act as potential breast CSC inhibitors. In the third study, the Ras FRET-biosensor principle was used to investigate membrane anchorage and nanoclustering of myristoylated proteins such as heterotrimeric G-proteins, Yes- and Src-kinases. Furthermore, Yes-biosensor was validated to be a suitable platform for performing chemical and genetic screens to identify myristoylation inhibitors. The results of this thesis demonstrate the potential of the Ras-derived FRETbiosensor platform to differentiate and identify Ras-isoform specfic inhibitors. The results also highlight that most of the inhibitors identified predominantly perturb Ras subcellular distribution and membrane organization through some novel and yet unknown mechanisms. The results give new insights into the role of Ras nanoclusters as promising new molecular targets in cancer and in stem cells.