Cross-species analysis of genetically engineered mouse models of MAPK-driven colorectal cancer identifies hallmarks of the human disease

Effective treatment options for advanced colorectal cancer (CRC) are limited, survival rates are poor and this disease continues to be a leading cause of cancer-related deaths worldwide. Despite being a highly heterogeneous disease, a large subset of individuals with sporadic CRC typically harbor relatively few established ‘driver’ lesions. Here, we describe a collection of genetically engineered mouse models (GEMMs) of sporadic CRC that combine lesions frequently altered in human patients, including well-characterized tumor suppressors and activators of MAPK signaling. Primary tumors from these models were profiled, and individual GEMM tumors segregated into groups based on their genotypes. Unique allelic and genotypic expression signatures were generated from these GEMMs and applied to clinically annotated human CRC patient samples. We provide evidence that a Kras signature derived from these GEMMs is capable of distinguishing human tumors harboring KRAS mutation, and tracks with poor prognosis in two independent human patient cohorts. Furthermore, the analysis of a panel of human CRC cell lines suggests that high expression of the GEMM Kras signature correlates with sensitivity to targeted pathway inhibitors. Together, these findings implicate GEMMs as powerful preclinical tools with the capacity to recapitulate relevant human disease biology, and support the use of genetic signatures generated in these models to facilitate future drug discovery and validation efforts.KEY WORDS: KRAS, BRAF, MAPK, Colorectal cancer, GEMM, Genomic signatures     

Mutation of the Arabidopsis LON2 peroxisomal protease enhances pexophagy

Peroxisomes are critical organelles housing various, often oxidative, reactions. Pexophagy, the process by which peroxisomes are selectively targeted for destruction via autophagy, is characterized in yeast and mammals but had not been reported in plants. In this article, we describe how the peroxisome-related aberrations of a mutant defective in the LON2 peroxisomal protease are suppressed when autophagy is prevented by mutating any of several key autophagy-related (ATG) genes. Our results reveal that plant peroxisomes can be degraded by selective autophagy and suggest that pexophagy is accelerated when the LON2 protease is disabled.     

AnxA5 reduces plaque inflammation of advanced atherosclerotic lesions in apoE−/− mice

Annexin A5 (AnxA5) exerts anti‐inflammatory, anticoagulant and anti‐apoptotic effects through binding cell surface expressed phosphatidylserine. The actions of AnxA5 on atherosclerosis are incompletely understood. We investigated effects of exogenous AnxA5 on plaque morphology and phenotype of advanced atherosclerotic lesions in apoE^?/? mice. Advanced atherosclerotic lesions were induced in 12 weeks old Western type diet fed apoE^?/? mice using a collar placement around the carotid artery. After 5 weeks mice were injected either with AnxA5 (n = 8) or vehicle for another 4 weeks. AnxA5 reduced plaque macrophage content both in the intima (59% reduction, P < 0.05) and media (73% reduction, P < 0.01) of advanced atherosclerotic lesions of the carotid artery. These findings corroborated with advanced lesions of the aortic arch, where a 67% reduction in plaque macrophage content was observed with AnxA5 compared to controls (P < 0.01). AnxA5 did not change lesion extension, plaque apoptosis, collagen content, smooth muscle cell content or acellular plaque composition after 4 weeks of treatment as determined by immunohistochemistry in advanced carotid lesions. In vitro, AnxA5 exhibited anti‐inflammatory effects in macrophages and a flow chamber based assay demonstrated that AnxA5 significantly inhibited capture, rolling, adhesion as well as transmigration of peripheral blood mononuclear cells on a TNF‐α‐activated endothelial cell layer. In conclusion, short‐term treatment with AnxA5 reduces plaque inflammation of advanced lesions in apoE^?/? mice likely through interfering with recruitment and activation of monocytes to the inflamed lesion site. Suppressing chronic inflammation by targeting exposed phosphatidylserine may become a viable strategy to treat patients suffering from advanced atherosclerosis.     

Treadmill Exercise Induces Neutrophil Recruitment into Muscle Tissue in a Reactive Oxygen Species-Dependent Manner. An Intravital Microscopy Study

Intense exercise is a physiological stress capable of inducing the interaction of neutrophils with muscle endothelial cells and their transmigration into tissue. Mechanisms driving this physiological inflammatory response are not known. Here, we investigate whether production of reactive oxygen species is relevant for neutrophil interaction with endothelial cells and recruitment into the quadriceps muscle in mice subjected to the treadmill fatiguing exercise protocol. Mice exercised until fatigue by running for 56.3±6.8 min on an electric treadmill. Skeletal muscle was evaluated by intravital microscopy at different time points after exercise, and then removed to assess local oxidative stress and histopathological analysis. We observed an increase in plasma lactate and creatine kinase (CK) concentrations after exercise. The numbers of monocytes, neutrophils, and lymphocytes in blood increased 12 and 24 hours after the exercise. Numbers of rolling and adherent leukocytes increased 3, 6, 12, and 24 hours post-exercise, as assessed by intravital microscopy. Using LysM-eGFP mice and confocal intravital microscopy technology, we show that the number of transmigrating neutrophils increased 12 hours post-exercise. Mutant gp91^phox-/- (non-functional NADPH oxidase) mice and mice treated with apocynin showed diminished neutrophil recruitment. SOD treatment promoted further adhesion and transmigration of leukocytes 12 hours after the exercise. These findings confirm our hypothesis that treadmill exercise increases the recruitment of leukocytes to the postcapillary venules, and NADPH oxidase-induced ROS plays an important role in this process.     

Chirally-Modifiedoligonucleotides and the Control of Gene Expression. The Case of L-DNAS And-RNAS

Abstract

The affinity of L-DNAs, L-RNAs and L/D-DNAs for homopurine homopyrimidine d.s. D-DNA and s.s. D-RNA was probed by gel electrophoresis and CD spectroscopy. It was found that the L-modified oligomers do not bind to d.s. DNA and to natural RNA that contains all four natural bases. Thus they cannot be used, in general, for the control of gene expression according to the antigene and antisense methodologies. Heterochiral complexes with 1:1 stoichiometry and low thermal stability are formed, instead, by homopurinic L-RNA or L/D-DNA and homopyrimidinic L-RNA with the W/C complementary natural RNA sequences.     

Study of Zn,Cu and Pb content in plants and contaminated soils in Sardinia

Trace elements in soils exist as components of several different fractions. We have analyzed the correlation between total and extractable (EDTA, calcium chloride and deionized water) Zn, Pb and Cu concentrations in soils and the concentration of these elements in plant leaves. Soil and plant samples have been taken from Sulcis-Iglesiente (Sardinia), an area rich in mining tailings. This has made that the concentrations of the trace element under study in soils were varied. Three plants have been studied: Dittrichia viscosa, Cistus salviifolius, and Euphorbia pithyusa subsp. cupanii. Soil samples beneath each of them at depths of 0–30 and 30–60 cm have been considered. The highest concentration of trace elements in the leaves of the studied species has been found for Zn. The calcium carbonate content and the crystalline and amorphous forms of iron in the soil have determined the concentration of metal in plant leaves. The soil concentrations that have been found with the extraction methods are uncorrelated with Pb and Cu concentrations in plants, but Zn is correlated with the fraction extracted with EDTA and calcium chloride. The concentrations of trace metals in plants are most closely related to the soil contents of CaCO₃, electrical conductivity, Fe_ox, and Fe_dc.     

Loss detection of aquatic habitats in Italian volcanic lakes using historical data

Italian volcanic lakes represent an important hotspot of aquatic plant diversity. By comparing original data collected in 2009/10 with historical data, we detected species extinction and loss of habitats 3140 and 3150 (Habitats Directive 92/43/EEC). The speed of change detected claim for frequent monitoring and rapid management interventions.     

Emission of isoprenoids from natural vegetation in the Beijing region (Northern China)

ABSTRACT

A survey was conducted to identify plants emitting isoprenoids in the Beijing area which could potentially contribute to smog episodes when combined with anthropogenic pollutants. The emission pattern was similar to that observed in the previously surveyed boreal ecosystems (Europe, North America). Most deciduous oaks are strong isoprene emitters; however, some of them do not emit isoprenoids and are therefore more suitable for the urban environment of Beijing. No emission of monoterpenes was found in Chinese oaks, and this trait seems therefore confined to the Mediterranean environment. The emission of isoprene was found in poplars and in some of the bamboo widespread in city parks and in the riparial vegetation surrounding the city. Chinese pine species emit monoterpenes when wounded, and the emission is not qualitatively different among species. Pinus tabulaeformis, one of the most important trees in China, is a low emitter compared to other pine species.