Differential chemosensitivity to antifolate drugs between RAS and BRAF melanoma cells.

BACKGROUND: The importance of the genetic background of cancer cells for the individual susceptibility to cancer treatments is increasingly apparent. In melanoma, the existence of a BRAF mutation is a main predictor for successful BRAF-targeted therapy. However, despite initial successes with these therapies, patients relapse within a year and have to move on to other therapies. Moreover, patients harbouring a wild type BRAF gene (including 25% with NRAS mutations) still require alternative treatment such as chemotherapy. Multiple genetic parameters have been associated with response to chemotherapy, but despite their high frequency in melanoma nothing is known about the impact of BRAF or NRAS mutations on the response to chemotherapeutic agents. METHODS: Using cell proliferation and DNA methylation assays, FACS analysis and quantitative-RT-PCR we have characterised the response of a panel of NRAS and BRAF mutant melanoma cell lines to various chemotherapy drugs, amongst them dacarbazine (DTIC) and temozolomide (TMZ) and DNA synthesis inhibitors. RESULTS: Although both, DTIC and TMZ act as alkylating agents through the same intermediate, NRAS and BRAF mutant cells responded differentially only to DTIC. Further analysis revealed that the growth-inhibitory effects mediated by DTIC were rather due to interference with nucleotide salvaging, and that NRAS mutant melanoma cells exhibit higher activity of the nucleotide synthesis enzymes IMPDH and TK1. Importantly, the enhanced ability of RAS mutant cells to use nucleotide salvaging resulted in resistance to DHFR inhibitors. CONCLUSION: In summary, our data suggest that the genetic background in melanoma cells influences the response to inhibitors blocking de novo DNA synthesis, and that defining the RAS mutation status could be used to stratify patients for the use of antifolate drugs

Spatial control of Cdc42 signalling by a GM130-RasGRF complex regulates polarity and tumorigenesis

The small GTPase Cdc42 is a key regulator of polarity, but little is known in mammals about its spatial regulation and the relevance of spatial Cdc42 pools for polarity. Here we report the identification of a GM130-RasGRF complex as a regulator of Cdc42 at the Golgi. Silencing GM130 results in RasGRF-dependent inhibition of the Golgi pool of Cdc42, but does not affect Cdc42 at the cell surface. Furthermore, active Cdc42 at the Golgi is important to sustain asymmetric front-rear Cdc42-GTP distribution in directionally migrating cells. Concurrent to Cdc42 inhibition, silencing GM130 also results in RasGRF-dependent Ras-ERK pathway activation. Moreover, depletion of GM130 is sufficient to induce E-cadherin downregulation, indicative of a loss in cell polarity and epithelial identity. Accordingly, GM130 expression is frequently lost in colorectal and breast cancer patients. These findings establish a previously unrecognized role for a GM130-RasGRF-Cdc42 connection in regulating polarity and tumorigenesis

PDL1 Signals through Conserved Sequence Motifs to Overcome Interferon-Mediated Cytotoxicity

PDL1 blockade produces remarkable clinical responses, thought to occur by T cell reactivation through prevention of PDL1-PD1 T cell inhibitory interactions. Here, we find that PDL1 cell-intrinsic signaling protects cancer cells from interferon (IFN) cytotoxicity and accelerates tumor progression. PDL1 inhibited IFN signal transduction through a conserved class of sequence motifs that mediate crosstalk with IFN signaling. Abrogation of PDL1 expression or antibody-mediated PDL1 blockade strongly sensitized cancer cells to IFN cytotoxicity through a STAT3/caspase-7-dependent pathway. Moreover, somatic mutations found in human carcinomas within these PDL1 sequence motifs disrupted motif regulation, resulting in PDL1 molecules with enhanced protective activities from type I and type II IFN cytotoxicity. Overall, our results reveal a mode of action of PDL1 in cancer cells as a first line of defense against IFN cytotoxicity

FOSL1 promotes cholangiocarcinoma via transcriptional effectors that could be therapeutically targeted

Understanding the molecular mechanisms involved in cholangiocarcinoma (bile duct cancer) development and progression stands as a critical step for the development of novel therapies. Through an inter-species approach, this study provides evidence of the clinical and functional role of the transcription factor FOSL1 in cholangiocarcinoma. Moreover, we report that downstream effectors of FOSL1 are susceptible to pharmacological inhibition, thus providing new opportunities for therapeutic intervention

The expression of monocarboxylate transporters in thyroid carcinoma can be associated with the morphological features of BRAF (V600E) mutation

BRAF (V600E) mutation, usually performed by DNA techniques, is one of the most common diagnostic markers in papillary thyroid carcinoma. Few papers have demonstrated that plump cells (eosinophilic cytoplasms and papillary thyroid carcinoma nuclei) and peculiar sickle-shaped nuclei represent morphological features of BRAF (V600E) on papillary thyroid carcinomas. These features seem to be linked to glycolytic phenotype whereby monocarboxylate transporters 1-4 are hypothesized to have a dominant role as lactate transporters. We investigated the association between these morphological features and monocarboxylate transporters 1 and 4 in 48 cyto-histological samples diagnosed as "positive for malignancy-favoring papillary thyroid carcinoma". These cases were processed with liquid-based cytology and underwent BRAF (V600E) mutational analysis (pyrosequencing) on liquid-based cytology and monocarboxylate transporters immunostaining on histology. The expression of monocarboxylate transporter 1, monocarboxylate transporter 4, glucose trasporter-1 and carbonic anhidrase were scored semi-quantitatively with expression from 0 to 3+ (strong positivity). The 33 mutated and 15 wild type cases showed 100 % cyto-histological concordance. The cytological evaluation revealed plump cells and sickle nuclear shape in 100 % mutated cases. Monocarboxylate transporter 1 yielded 76 % positivity in the mutated cases especially in both the plump cells and sickle-shaped nuclei, whereas the wild types showed 13.3 % positive monocarboxylate transporter 1 (p = 0.00013). Monocarboxylate transporter 4 resulted in 100 % positivity in mutated and 40 % in wild types (p 0.05). This is the first report analyzing the association between monocarboxylate transporter expression and the morphological features of BRAF (V600E) mutated papillary thyroid carcinomas suggesting the possible involvement of lactate in the morphological features.info:eu-repo/semantics/publishedVersio

WNT/β-catenin signaling regulates mitochondrial activity to alter the oncogenic potential of melanoma in a PTEN-dependent manner

Aberrant regulation of WNT/β-catenin signaling has a crucial role in the onset and progression of cancers, where the effects are not always predictable depending on tumor context. In melanoma, for example, models of the disease predict differing effects of the WNT/β-catenin pathway on metastatic progression. Understanding the processes that underpin the highly context-dependent nature of WNT/β-catenin signaling in tumors is essential to achieve maximal therapeutic benefit from WNT inhibitory compounds. In this study, we have found that expression of the tumor suppressor, phosphatase and tensin homolog deleted on chromosome 10 (PTEN), alters the invasive potential of melanoma cells in response to WNT/β-catenin signaling, correlating with differing metabolic profiles. This alters the bioenergetic potential and mitochondrial activity of melanoma cells, triggered through regulation of pro-survival autophagy. Thus, WNT/β-catenin signaling is a regulator of catabolic processes in cancer cells, which varies depending on the metabolic requirements of tumors

AI is a viable alternative to high throughput screening: a 318-target study

: High throughput screening (HTS) is routinely used to identify bioactive small molecules. This requires physical compounds, which limits coverage of accessible chemical space. Computational approaches combined with vast on-demand chemical libraries can access far greater chemical space, provided that the predictive accuracy is sufficient to identify useful molecules. Through the largest and most diverse virtual HTS campaign reported to date, comprising 318 individual projects, we demonstrate that our AtomNet® convolutional neural network successfully finds novel hits across every major therapeutic area and protein class. We address historical limitations of computational screening by demonstrating success for target proteins without known binders, high-quality X-ray crystal structures, or manual cherry-picking of compounds. We show that the molecules selected by the AtomNet® model are novel drug-like scaffolds rather than minor modifications to known bioactive compounds. Our empirical results suggest that computational methods can substantially replace HTS as the first step of small-molecule drug discovery

The rip current hazard in Costa Rica

Deaths from rip currents are a major hazard on global beaches, but few data are available at a national scale to support the development of appropriate intervention programs. Analysis of data from the Judicial Investigation Organization of Costa Rica indicates that drowning is the leading cause of violent death in the country, with 1,391 drownings between 2001 and 2012. Approximately 590 of those drownings occurred in a marine environment and are listed as being the result of rip currents. A majority (64 %) of the drownings attributed to rip currents involved victims from Costa Rica and tended to involve young single male students at beaches within a relatively short drive from San Jose on weekends and on national holidays. In comparison, the majority of foreign drowning victims were older males from the USA and Nicaragua with no statistically significant variation in the number of those drownings by day of the week. Predicted wave forcing and measured sediment characteristics suggest that the majority of drownings are associated with a transverse bar and rip (TBR) morphology with quasi-steady rip channels close to the beach. The interannual variation in both local and foreign drownings exhibits a statistically significant dependency on the variation in wave height with the Pacific-North American (PNA) Oscillation. Specifically, drownings peak during the negative phase of the PNA when wave heights are significantly smaller, which may reflect a tendency for beach users to enter calm water when the beach tends to have a TBR morphology. Further study is required to characterize the rip hazard at the most popular beaches on the Pacific and Caribbean coasts and to determine the level of rip knowledge by both local and foreign beach users. © 2015, Springer Science+Business Media Dordrecht

The relative influence of lithology and weathering in shaping shore platforms along the coastline of the Gulf of La Spezia (NW Italy) as revealed by rock strength

Along the rock coasts of the Gulf of La Spezia, which are characterised by a Mediterranean microtidal environment, a limited number of small rock platforms are scattered, constrained in elevation within 5 m above present-day sea level. This work deals with a number of these rock platforms, formed in different rock types (one in limestone and two in dolomite), that show differences in their morphology. This paper aims to provide a quantitative examination of why there are morphological differences between platforms in this region. To achieve this purpose, factors controlling platform morphology and the processes acting on them are investigated through a comparative analysis of rock strength. Rebound values, obtained testing rock surfaces with the Schmidt hammer, were compared between different platforms and between different sectors of the same platform. Each platform was subdivided into two parts based on visual difference in rock surface colour, characterised by differences in occurrence of weathering microforms and bioerosive agents. Rebound values in the lower part of the platforms proved to be lower than in the upper part, providing quantitative assessment of the occurrence of weathering acting to different extents in the upper and lower part of the shore platforms (weathering degraded rock strength in the lower part by about 15%). It was demonstrated that on the upper part of platforms, displaying moderate evidence of physical and biological weathering, lithology significantly influences the rock strength. On the portion of platforms closer to sea level, instead, differential exposure histories of the same rock type in the same environmental setting can yield statistically significant variations in rock strength values. Thus, it is clear that in the lower part of the investigated platforms, the degree of weathering has strong bearing on rock strength, and that variations in rock strength are not solely due to lithology. According to the results of this work, experimental values of rock strength of platforms in the study area depend both on the rock type and on physical weathering due to frequently repeated wetting and drying and bioerosion. Lithology is then an important factor controlling platform shape and weathering is an important process operating on them

A new approach for the study of the coast indentation index

The indentation index, which is the ratio between the real length of a coast and its Euclidean length, is a parameter applied in order to characterize rock coasts and to study their evolution. Rather than subjectively selecting two or more sectors of a rock coast where to evaluate, compare and contrast it, in this paper it is suggested to consider a same coastline and to split it into several adjacent tracts of equal Euclidean length within which the indentation index can be calculated. By digitizing the coastline in a GIS environment, it becomes possible to test several Euclidean length values on a same coastline, each time obtaining a different spatial variability of the indentation index. The best length values that maximize the spatial variability of the indentation index are those that determine an indentation index pattern characterized by a high variance and a low spatial autocorrelation. The spatial distribution of indentation index can eventually be commented at the light of the known littoral forces acting on the studied coast. When more than one Euclidean length value is found to maximize the variability of indentation index within the same coast, it is likely that there are one or more littoral forces acting or interacting differently at different scales