Correction of aberrant FGFR1 RNA splicing through blocking intronic regulatory elements

Analysis of the human genome has revealed that more than 74% of human genes undergo alternative RNA splicing. Aberrations in alternative RNA splicing have been associated with several human disorders, including cancer. ^ We studied the aberrant expression of alternative RNA splicing isoforms of the Fibroblast Growth Factor Receptor 1 (FGFR1) gene in a human glioblastoma cancer model. Normal glial cells express the FGFR1α, which contains three extracellular domains. In tumors the most abundant isoform is the FGFR1β, which lacks the first extracellular domain due to the skipping of a single exon, termed alpha. The skipping of the α-exon is regulated by two intronic silencing sequences within the precursor mRNA. Since we observed no mutations on these elements in tumor cells, we hypothesized that the over-expression of regulatory proteins that recognize these sequences is responsible for the aberrant expression of splicing isoforms. Hence, we blocked the formation of protein complexes on the ISS using antisense RNA oligonucleotides in vitro. We also evaluated the impact of the ISS antisense oligonucleotides on the endogenous FGFR1 splicing, in a glioblastoma cell model. By targeting intronic regulatory elements we were able to increase the level of alpha exon inclusion up to 90% in glioblastoma cells. The effect was dose dependent, sequence specific and reproducible in glioblastoma and other cancer cells, which also exhibit an alpha exon skipping phenotype. Targeting FGFR1 endogenous ISS1 and ISS2 sequences did not have an additive or synergistic effect, which suggest a regulatory splicing mechanism that requires the interaction of complexes formed on these elements. An increase in the levels of the FGFR1α isoform resulted in a reduction in cell invasiveness. Also, a significant increase in the levels of caspase 3/7 activities, which is indicative of an elevation in apoptosis levels, suggests that expression of FGFR1β might be relevant for tumor survival. These studies demonstrate that it is possible to prevent aberrant expression of exon skipping events through the targeting of intronic regulatory elements, providing an important new therapeutic tool for the correction of human disease caused by alternative RNA splicing. ^

J. Ebbeler, Disciplining Christians: Correction and Community in Augustine’s Letters, Oxford University Press, New York 2012, 254 pp. ISBN 9780195372564.

Fil: Correa, Soledad. Universidad Nacional de Rosario

High resolution bathymetric compilation for Potter Cove, WAP, Antarctica, with links to data in ArcGIS format

The bathymetry raster with a resolution of 5 m x 5 m was processed from unpublished single beam data from the Argentine Antarctica Institute (IAA, 2010) and multibeam data from the United Kingdom Hydrographic Office (UKHO, 2012) with a cell size of 5 m x 5 m. A coastline digitized from a satellite image (DigitalGlobe, 2014) supplemented the interpolation process. The 'Topo to Raster' tool in ArcMap 10.3 was used to merge the three data sets, while the coastline represented the 0-m-contour to the interpolation process ('contour type option').

Updated bathymetric chart of the East Aral Sea

The Aral Sea is located in an arid region with much sand and salt deposits in the surrounding barren open land. Hence, significant displacements of sediments into the lakebed by the action of wind, water, gravity, or snow are likely. The bathymetry of the lake was last observed in the 1960s, and within the last half century, the structure of the lakebed has changed. Based on satellite observations of the temporal changes of shoreline (Landsat optical remote sensing) and water level (multi-mission satellite altimetry) over more than one decade an updated bathymetric chart for the East Basin of the Aral Sea has been generated. During this time, the geometry of the shallow East Basin experienced strong fluctuations due to the occurrence of periods of drying and strong inflow. By the year 2014 the East Basin fell dry. The dynamic behavior of the basin allowed for estimating the lake's bathymetry from a series of satellite-based information. The river mouth made its impression in the present East Aral Sea, because its shrinking led to the inflow of much sediment into the lake's interior. In addition, salt deposits along the shorelines increased the corresponding elevation, a phenomenon that is more pronounced in the reduced lakebed because of increased salinity. It must be noted that height estimates from satellite altimetry were only possible down to a minimum elevation of 27 m above sea level due to a lack of reliable altimetry data over the largely reduced water surface. In order to construct a complete bathymetric chart of the lakebed of the East Aral Sea heights below 27 m were obtained solely from Landsat optical images following the SRB (Selected Region Boundary) approach as described by Singh et al. (2015).