Lithostratigraphy and age of pre-Late Weichselian sediments in the Suupohja area, western Finland

Different types of laterally extensive sand- and gravel-dominated deposits, up to several tens of metres thick, were investigated in the Suupohja area of western Finland. The studied sediments were deposited in glacial, ice-marginal, glaciofluvial, sea or lake, littoral and terrestrial environments during several glacial-non-glacial cycles. Seventeen pre-Late Weichselian and three Late Weichselian/Holocene sedimentary units were identified. These were divided into ten formally and two informally defined formations that were together termed the Suupohja Group. Every unit are nevertheless not detectable throughout the study area. The informally defined “Karhukangas lower deposits” represent the lowest units in the Suupohja Group. The Karhukangas lower deposits with 5 till units, 3 glaciolacustrine/-marine units and 2 sand units, were interpreted as having been deposited during possibly four glacial-non-glacial cycles before the Late Pleistocene Subepoch (MIS 6 or earlier). The Kankalo Sand above the Karhukangas lower deposits comprises glaciofluvial and aeolian sands of Late Saalian, Eemian or Early Weichselian origin (MIS 6–MIS 5c). The Kariluoma Till above the Kankalo Sand was possibly deposited during the Late Saalian glacial advance, although an Early Weichselian origin is also possible. The Harrinkangas Formation, with glaciofluvial and quiet-water sediments, is interpreted as having been deposited during the Late Saalian and Eemian Stages (MIS 6–MIS 5e). The uppermost units in the deposits studied, the Kodesjärvi Formation (shore deposit), Isojoki Sand (aeolian), Rävåsen Formation (glaciofluvial), Vanhakylä Formation (shore line deposit), Dagsmark Till and Kauhajoki Till, were deposited during the Weichselian Stage (MIS 5d–MIS 2). In addition, Early Holocene (MIS 1) eskers without till cover were informally termed the “Holocene esker deposits”. The Lumikangas Formation represents gravelly shore deposits formed in the Holocene Epoch, when these areas last emerged from the sea. The first Weichselian ice expansion possibly reached the western part of Suupohja in the Early Weichselian Substage (MIS 5d?), but it did not expand further to the east. The second Weichselian glaciation of relatively short duration occupied the southern part of Finland in the later part of Middle Weichselian (MIS 3). Thus, the southern half of the country remained ice-free for the majority (~65–75%) of the Weichselian Stage. Instead, both humid temperate and periglacial conditions alternated. In the initial part of Middle Weichselian, this area was partly submerged, which indicates eastward expansion of the Scandinavian ice sheet(s), depressing the lithosphere. The exceptionally thick sediment cover, multiple lithofacies, relict landscape and preserved preglacially weathered bedrock are evidence of weak glacial erosion in the Suupohja area during the latest as well as earlier glaciations, making this area one of the key areas in Quaternary research.

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.