Identification and pharmacological inactivation of the MYCN gene network as a therapeutic strategy for neuroblastic tumor cells

This research was originally published in Journal of Biological Chemistry. Olesya Chayka, Cosimo Walter D’Acunto, Odette Middleton, Maryam Arab, and Arturo Sala. Identification and Pharmacological Inactivation of the MYCN Gene Network as a Therapeutic Strategy for Neuroblastic Tumor Cells. Journal of Biological Chemistry. 2015; Vol 290 (4) :pp. 2198 - 2212. © the American Society for Biochemistry and Molecular Biology.This article has been made available through the Brunel Open Access Publishing Fund.The MYC family of transcription factors consists of three well characterized members, c-MYC, L-MYC, and MYCN, deregulated in the majority of human cancers. In neuronal tumors such as neuroblastoma, MYCN is frequently activated by gene amplification, and reducing its expression by RNA interference has been shown to promote growth arrest and apoptosis of tumor cells. From a clinical perspective, RNA interference is not yet a viable option, and small molecule inhibitors of transcription factors are difficult to develop. We therefore planned to identify, at the global level, the genes interacting functionally with MYCN required to promote fitness of tumor cells facing oncogenic stress. To find genes whose inactivation is synthetically lethal to MYCN, we implemented a genome-wide approach in which we carried out a drop-out shRNA screen using a whole genome library that was delivered into isogenic neuroblastoma cell lines expressing or not expressing MYCN. After the screen, we selected for in-depth analysis four shRNAs targeting AHCY, BLM, PKMYT1, and CKS1B. These genes were chosen because they are directly regulated by MYC proteins, associated with poor prognosis of neuroblastoma patients, and inhibited by small molecule compounds. Mechanistically, we found that BLM and PKMYT1 are required to limit oncogenic stress and promote stabilization of the MYCN protein. Cocktails of small molecule inhibitors of CKS1B, AHCY, BLM, and PKMYT1 profoundly affected the growth of all neuroblastoma cell lines but selectively caused death of MYCN-amplified cells. Our findings suggest that drugging the MYCN network is a promising avenue for the treatment of high risk, neuroblastic cancers.SPARKS and the Neuroblastoma Society

Frizzled receptor 6 marks rare, highly tumourigenic stem-like cells in mouse and human neuroblastomas

Copyright © 2011 Cantilena et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 3.0 License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. The article was made available through the Brunel Open Access Publishing Fund.Wnt signalling is an important component of vertebrate development, required for specification of the neural crest. Ten Wnt receptors [Frizzled receptor 1-10 (Fzd1-10)] have been identified so far, some of which are expressed in the developing nervous system and the neural crest. Here we show that expression of one such receptors, Fzd6, predicts poor survival in neuroblastoma patients and marks rare, HIF1/2 α-positive cells in tumour hypoxic areas. Fzd6 positive neuroblastoma cells form neurospheres with high efficiency, are resistant to doxorubicin killing and express high levels of mesenchymal markers such as Twist1 and Notch1. Expression of Fzd6 is required for the expression of genes of the noncanonical Wnt pathway and the spheres forming activity. When transplanted into immunodeficient mice, neuroblastoma cells expressing the Fzd6 marker grow more aggressively than their Fzd6 negative counterparts. We conclude that Fzd6 is a new surface marker of aggressive neuroblastoma cells with stem cell-like features.This work was sponsored by the Wellcome Trust, the RICC cancer fund, SPARKS, the Italian Association for Cancer Research, Regione Liguria and the Italian Ministry of Health

Physical interaction between MYCN oncogene and polycomb repressive complex 2 (PRC2) in neuroblastoma: Functional and therapeutic implications

This article is made available through the Brunel Open Access Publishing Fund. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.CLU (clusterin) is a tumor suppressor gene that we have previously shown to be negatively modulated by the MYCN proto-oncogene, but the mechanism of repression was unclear. Here, we show that MYCN inhibits the expression of CLU by direct interaction with the non-canonical E box sequence CACGCG in the 5′-flanking region. Binding of MYCN to the CLU gene induces bivalent epigenetic marks and recruitment of repressive proteins such as histone deacetylases and Polycomb members. MYCN physically binds in vitro and in vivo to EZH2, a component of the Polycomb repressive complex 2, required to repress CLU. Notably, EZH2 interacts with the Myc box domain 3, a segment of MYC known to be essential for its transforming effects. The expression of CLU can be restored in MYCN-amplified cells by epigenetic drugs with therapeutic results. Importantly, the anticancer effects of the drugs are ablated if CLU expression is blunted by RNA interference. Our study implies that MYC tumorigenesis can be effectively antagonized by epigenetic drugs that interfere with the recruitment of chromatin modifiers at repressive E boxes of tumor suppressor genes such as CLU.SPARKS, The Neuroblastoma Society, a Wellcome Trust grant (to A. S.), and the Italian Association for Cancer Research

XMM-Newton observations of Nova Sgr 1998

We report on X-ray observations of Nova Sagittarius 1998 (V4633 Sgr), performed with XMM-Newton at three different epochs, 934, 1083 and 1265 days after discovery. The nova was detected with the EPIC cameras at all three epochs, with emission spanning the whole energy range from 0.2 to 10 keV. The X-ray spectra do not change significantly at the different epochs, and are well fitted for the first and third observations with a multi-temperature optically thin thermal plasma, while lower statistics in the second observations lead to a poorer fit. The thermal plasma emission is most probably originated in the shock heated ejecta, with chemical composition similar to that of a CO nova. However, we can not completely rule out reestablished accretion as the origin of the emission. We also obtain upper limits for the temperature and luminosity of a potential white dwarf atmospheric component, and conclude that hydrogen burning had already turned-off by the time of our observations.Comment: 18 pages, 3 figures. Accepted in Astrophysical Journa

Charge localization at the interface between La1-xSrxMnO3 and the infinite layers cuprate CaCuO2

(CaCuO2)m/(La0.7Sr0.3MnO3)n superlattices, consisting of the infinite layers cuprate CaCuO2 and the optimally doped manganite La1-xSrxMnO3, were grown by pulsed laser deposition. The transport properties are dominated by the manganite block. X-Ray Absorption spectroscopy measurements show a clear evidence of an orbital reconstruction at the interface, ascribed to the hybridization between the Cu 3d3z2-r2 and the Mn 3d3z2-r2 orbitals via interface apical oxygen ions. Such a mechanism localizes holes at the interfaces, thus preventing charge transfer to the CaCuO2 block. Some charge (holes) transfer occurs toward the La0.7Sr0.3MnO3 block in strongly oxidized superlattices, contributing to the suppression of the magnetotransport properties.Comment: 20 pages, 6 figure

Hydrologic Niches Explain Species Coexistence and Abundance in a Shrub-Steppe System

Differences in vertical root distributions are often assumed to create resource uptake tradeoffs that determine plant growth and coexistence. Yet, most plant roots are in shallow soils, and data linking root distributions with resource uptake and plant abundances remain elusive. Here we used a tracer experiment to describe the vertical distribution of absorptive roots of dominant species in a shrub‐steppe ecosystem. To describe how these different rooting distributions affected water uptake in wet and dry soils across a growing season, we used a soil water movement model. Root traits were then correlated with plant landscape abundances. Deeper root distributions extracted more soil water, had larger unique hydrological niches and were more abundant on the landscape. Though most (\u3e 50%) root biomass and tracer uptake occurred in shallow soils (0–32 cm), the depth of 50% of tracer uptake varied from 11 to 32 cm across species and species with deeper rooting distributions were more abundant on the landscape (R2 = 0.95). The water flow model revealed that deeper rooting distributions should extract more soil water (i.e., a range of 60 to 113 mm of soil water) because shallow roots were often in dry soils. These potential water uptake values were tightly correlated with species’ abundances on the landscape (R2 = 0.90). Finally, each species’ rooting distribution demonstrated a depth and time at which it could extract more soil water than any other rooting distribution, and the size of these unique hydrological niches indices was also well correlated with species’ abundances (R2 = 0.89). Synthesis. Our results demonstrate not only a correlation between root distributions and species abundance, but also the mechanism through which differences in rooting distributions can determine resource uptake and niche partitioning, even when most roots are found in shallow soils

Primary production of the central grassland region of the United States

Includes bibliographical references (pages 44-45).Aboveground net primary production of grasslands is strongly influenced by the amount and distribution of annual precipitation. Analysis of data collected at 9500 sites throughout the central United States confirmed the overwhelming importance of water availability as a control of production. The regional spatial pattern of production reflected the east-west gradient in annual precipitation. Lowest values of aboveground net primary production were observed in the west and highest values in the east. This spatial pattern was shifted eastward during unfavorable years and westward during favorable years. Variability in production among years was maximum in northern New Mexico and southwestern Kansas and decreased towards the north and south. The regional pattern of production was largely accounted for by annual precipitation. Production at the site level was explained by annual precipitation, soil water-holding capacity, and an interaction term. Our results support the inverse texture hypothesis. When precipitation is 370 mm/yr

Phylogenetic analysis of grapevine cv. Ansonica growing on the island of Giglio, Italy, by AFLP and SSR markers

The geographic origin of the grapevine cultivar Ansonica (Vitis vinifera L.) grown on the island of Giglio, Tuscany, Italy, was investigated with molecular tools, i.e. AFLP and SSR (microsatellite) analysis. The scored polymorphic DNA bands were statistically analysed and the results were expressed in dendrograms showing the degree of genomic similarity among the tested cultivars: three Ansonica vines sampled in some of the oldest vineyards of the island, one Ansonica specimen grown in Tuscany, one Inzolia specimen grown in Sicily, 23 cultivars grown in different Mediterranean regions and 40 Greek cultivars. The results obtained from AFLP or SSR approaches led to equivalent conclusions: the three grapevines sampled in Giglio were indistinguishable and showed genetic similarity with cv. Ansonica grown in the Tuscan mainland and with cv. Inzolia from Sicily, Airen from Spain, Clairette from France and Roditis from Greece. The SSR analysis showed that Sideritis and Roditis have the highest genomic similarity with Ansonica among the 40 tested Greek cultivars. The molecular analysis gives conclusive evidence for the Greek origin of Ansonica grown on the island, as previously proposed on the basis of morphological and historical studies

Military spending and economic growth in China: a regime-switching analysis

This article has been made available through the Brunel Open Access Publishing Fund.This article investigates the impact of military spending changes on economic growth in China over the period 1953 to 2010. Using two-state Markov-switching specifications, the results suggest that the relationship between military spending changes and economic growth is state dependent. Specifically, the results show that military spending changes affect the economic growth negatively during a slower growth-higher variance state, while positively within a faster growth-lower variance one. It is also demonstrated that military spending changes contain information about the growth transition probabilities. As a policy tool, the results indicate that increases in military spending can be detrimental to growth during slower growth-higher growth volatility periods. © 2014 © 2014 The Author(s). Published by Taylor & Francis