Axitinib induces DNA damage response leading to senescence, mitotic catastrophe, and increased NK cell recognition in human renal carcinoma cells.

Tyrosine kinase inhibitors (TKIs) including axitinib have been introduced in the treatment of renal cell carcinoma (RCC) because of their anti-angiogenic properties. However, no evidence are presently available on a direct cytotoxic anti-tumor activity of axitinib in RCC.Herein we reported by western blot analysis that axitinib treatment induces a DNA damage response (DDR) initially characterized by γ-H2AX phosphorylation and Chk1 kinase activation and at later time points by p21 overexpression in A-498 and Caki-2 RCC cells although with a different potency. Analysis by immunocytochemistry for the presence of 8-oxo-7,8-dihydro-2'-deoxyguanosine in cellular DNA and flow cytometry using the redox-sensitive fluorescent dye DCFDA, demonstrated that DDR response is accompanied by the presence of oxidative DNA damage and reactive oxygen species (ROS) generation. This response leads to G2/M cell cycle arrest and induces a senescent-like phenotype accompanied by enlargement of cells and increased senescence-associated β-galactosidase activity, which are abrogated by N-acetyl cysteine (NAC) pre-treatment. In addition, axitinib-treated cells undergo to cell death through mitotic catastrophe characterized by micronucleation and abnormal microtubule assembly as assessed by fluorescence microscopy.On the other hand, axitinib, through the DDR induction, is also able to increase the surface NKG2D ligand expression. Accordingly, drug treatment promotes NK cell recognition and degranulation in A-498 RCC cells in a ROS-dependent manner.Collectively, our results indicate that both cytotoxic and immunomodulatory effects on RCC cells can contribute to axitinib anti-tumor activity

Capsaicin triggers autophagic cell survival which drives epithelial mesenchymal transition and chemoresistance in bladder cancer cells in an Hedgehog-dependent manner

Bladder cancer (BC) is a common urologic tumor characterized by high risk of recurrence and mortality. Capsaicin (CPS), used as an intravesical drug for overactive bladder, was demonstrated to induce cell death in different cancer cells including BC cells.Here we found that treatment of high-grade BC cells with high dose of CPS triggers autophagy. Infact, the CPS treatment alters the redox homeostasis by inducing production of radicals, mitochondrial depolarization, alterations of ADP/ATP ratio and activation of AMPK pathway stimulating the autophagic process in BC cells. The inhibition of autophagy, by using the specific inhibitor bafilomycin A or Beclin 1 knock-down, enhanced the CPS-induced cell death, demonstrating that CPS-induced autophagy acts as a pro-survival process in BC cells. By using PCR arrays and FACS analysis, we found that the CPS-treated BC cells displayed typical mesenchymal features of the epithelial mesenchymal transition (EMT) as elongated shape and over-expression of vimentin, α5 and β1 integrin subunits, integrin-like kinase and the anti-apoptotic Bcl-2 proteins. Moreover, we demonstrated that CPS treatment stimulates upregulation of Dhh/Ptch2/Zeb2 members of the Hedgehog signaling pathway, increases CD24, VEGFA and TIMP1 and decreases CD44 and ALCAM mRNA expression levels. By PTCH2 knock-down we found that the Hedgehog signaling pathway is involved in the CPS-induced autophagy and EMT phenotype.Finally, we also showed that the CPS-resistant EMT-positive BC cells displayed an increased drug-resistance to the cytotoxic effects of mitomycin C, gemcitabine and doxorubicine drugs commonly used in BC therapy

Axitinib induces senescence-associated cell death and necrosis in glioma cell lines: The proteasome inhibitor, bortezomib, potentiates axitinib-induced cytotoxicity in a p21(Waf/Cip1) dependent manner.

Glioblastoma is associated with a poor overall survival despite new treatment advances. Antiangiogenic strategies targeting VEGF based on tyrosine kinase inhibitors (TKIs) are currently undergoing extensive research for the treatment of glioma. Herein we demonstrated that the TKI axitinib induces DNA damage response (DDR) characterized by γ-H2AX phosphorylation and Chk1 kinase activation leading to G2/M cell cycle arrest and mitotic catastrophe in U87, T98 and U251 glioma cell lines. Moreover, we found that p21(Waf1/Cip1) increased levels correlates with induction of ROS and senescence-associated cell death in U87 and T98 cell lines, which are reverted by N-acetyl cysteine pretreatment. Conversely, U251 cell line showed a resistant phenotype in response to axitinib treatment, as evidenced by cell cycle arrest but no sign of cell death. The combinatorial use of axitinib with other therapies, with the aim of inhibiting multiple signaling pathways involved in tumor growth, can increase the efficiency of this TKI. Thus, we addressed the combined effects of axitinib with no toxic doses of the proteasome inhibitor bortezomib on the growth of U87 and T98 axitinib- sensitive and axitinib-resistant U251 cell lines. Compared to single treatments, combined exposure was more effective in inhibiting cell viability of all glioma cell lines, although with different cell death modalities. The regulation of key DDR and cell cycle proteins, including Chk1, γ-H2AX and p21(Waf1/Cip1) was also studied in glioma cell lines. Collectively, these findings provide new perspectives for the use of axitinib in combination with Bortezomib to overcome the therapy resistance in gliomas

MicroRNA-222 regulates muscle alternative splicing through Rbm24 during differentiation of skeletal muscle cells

A number of microRNAs have been shown to regulate skeletal muscle development and differentiation. MicroRNA-222 is downregulated during myogenic differentiation and its overexpression leads to alteration of muscle differentiation process and specialized structures. By using RNA-induced silencing complex (RISC) pulldown followed by RNA sequencing, combined with in silico microRNA target prediction, we have identified two new targets of microRNA-222 involved in the regulation of myogenic differentiation, Ahnak and Rbm24. Specifically, the RNA-binding protein Rbm24 is a major regulator of muscle-specific alternative splicing and its downregulation by microRNA-222 results in defective exon inclusion impairing the production of muscle-specific isoforms of Coro6, Fxr1 and NACA transcripts. Reconstitution of normal levels of Rbm24 in cells overexpressing microRNA-222 rescues muscle-specific splicing. In conclusion, we have identified a new function of microRNA-222 leading to alteration of myogenic differentiation at the level of alternative splicing, and we provide evidence that this effect is mediated by Rbm24 protei

Remote landslide mapping using a laser rangefinder binocular and GPS

We tested a high-quality laser rangefinder binocular coupled with a GPS receiver connected to a Tablet PC running dedicated software to help recognize and map in the field recent rainfall-induced landslides. The system was tested in the period between March and April 2010, in the Monte Castello di Vibio area, Umbria, Central Italy. To test the equipment, we measured thirteen slope failures that were mapped previously during a visual reconnaissance field campaign conducted in February and March 2010. For reference, four slope failures were also mapped by walking the GPS receiver along the landslide perimeter. Comparison of the different mappings revealed that the geographical information obtained remotely for each landslide by the rangefinder binocular and GPS was comparable to the information obtained by walking the GPS around the landslide perimeter, and was superior to the information obtained through the visual reconnaissance mapping. Although our tests were not exhaustive, we maintain that the system is effective to map recent rainfall induced landslides in the field, and we foresee the possibility of using the same (or similar) system to map landslides, and other geomorphological features, in other areas

Identification and mapping of recent rainfall-induced landslides using elevation data collected by airborne Lidar

International audienceA high resolution Digital Elevation Model with a ground resolution of 2 m×2 m (DEM2) was obtained for the Collazzone area, central Umbria, through weighted linear interpolation of elevation points acquired by Airborne Lidar Swath Mapping. Acquisition of the elevation data was performed on 3 May 2004, following a rainfall period that resulted in numerous landslides. A reconnaissance field survey conducted immediately after the rainfall period allowed mapping 70 landslides in the study area, for a total landslide area of 2.7×105 m2. Topographic derivative maps obtained from the DEM2 were used to update the reconnaissance landslide inventory map in 22 selected sub-areas. The revised inventory map shows 27% more landslides and 39% less total landslide area, corresponding to a smaller average landslide size. Discrepancies between the reconnaissance and the revised inventory maps were attributed to mapping errors and imprecision chiefly in the reconnaissance field inventory. Landslides identified exploiting the Lidar elevation data matched the local topography more accurately than the same landslides mapped using the existing topographic maps. Reasons for the difference include an incomplete or inaccurate view of the landslides in the field, an unfaithful representation of topography in the based maps, and the limited time available to map the landslides in the field. The high resolution DEM2 was compared to a coarser resolution (10 m×10 m) DEM10 to establish how well the two DEMs captured the topographic signature of landslides. Results indicate that the improved topographic information provided by DEM2 was significant in identifying recent rainfall-induced landslides, and was less significant in improving the representation of stable slopes

Landslide hazard assessment in the Collazzone area, Umbria, Central Italy

We present the results of the application of a recently proposed model to determine landslide hazard. The model predicts where landslides will occur, how frequently they will occur, and how large they will be in a given area. For the Collazzone area, in the central Italian Apennines, we prepared a multi-temporal inventory map through the interpretation of multiple sets of aerial photographs taken between 1941 and 1997 and field surveys conducted in the period between 1998 and 2004. We then partitioned the 79 square kilometres study area into 894 slope units, and obtained the probability of spatial occurrence of landslides by discriminant analysis of thematic variables, including morphology, lithology, structure and land use. For each slope unit, we computed the expected landslide recurrence by dividing the total number of landslide events inventoried in the terrain unit by the time span of the investigated period. Assuming landslide recurrence was constant, and adopting a Poisson probability model, we determined the exceedance probability of having one or more landslides in each slope unit, for different periods. We obtained the probability of landslide size, a proxy for landslide magnitude, by analysing the frequency-area statistics of landslides, obtained from the multi-temporal inventory map. Lastly, assuming independence, we determined landslide hazard for each slope unit as the joint probability of landslide size, of landslide temporal occurrence, and of landslide spatial occurrence

LANDSLIDE EVOLUTION PATTERN REVEALED BY MULTI-TEMPORAL DSMS OBTAINED FROM HISTORICAL AERIAL IMAGES

Abstract. Landslides are a widespread natural hazard that cause damages to people and to the built up environment. Accurate knowledge of landslide distribution is crucial to develop planning strategies, prevention and resilient communities worldwide. One of the most diffuse way of reporting landslides distribution in a territory is by preparing landslide inventory maps. Such a task is mostly accomplished by expert photo-interpretation of historical aerial photographs, which are an invaluable source of information because they portray the landscape when the anthropic pressure was lower than the present day, providing an observation of the landscape closer to the natural conditions. Despite such a common use of aerial photographs, they are poorly exploited to obtain quantitative measures to support landslide mapping activities. In this paper we present a comparison of two photogrammetric approaches to measure elevation changes in a 50-years period for an area densely affected by landslides in Southern Italy. The obtained results allowed to revisit the original expert mapping proving that such a method is a useful tool to support geomorphological mapping and to improve the overall accuracy of landslide inventories