Mitochondria-derived reactive oxygen species drive GANT61-induced mesothelioma cell apoptosis

Gli transcription factors of the Hedgehog (Hh) pathway have been reported to be drivers of malignant mesothelioma (MMe) cell survival. The Gli inhibitor GANT61 induces apoptosis in various cancer cell models, and has been associated directly with Gli inhibition. However various chemotherapeutics can induce cell death through generation of reactive oxygen species (ROS) but whether ROS mediates GANT61-induced apoptosis is unknown. In this study human MMe cells were treated with GANT61 and the mechanisms regulating cell death investigated. Exposure of MMe cells to GANT61 led to G1 phase arrest and apoptosis, which involved ROS but not its purported targets, GLI1 or GLI2. GANT61 triggered ROS generation and quenching of ROS protected MMe cells from GANT61-induced apoptosis. Furthermore, we demonstrated that mitochondria are important in mediating GANT61 effects: (1) ROS production and apoptosis were blocked by mitochondrial inhibitor rotenone; (2) GANT61 promoted superoxide formation in mitochondria; and (3) mitochondrial DNA-deficient LO68 cells failed to induce superoxide, and were more resistant to apoptosis induced by GANT61 than wild-type cells. Our data demonstrate for the first time that GANT61 induces apoptosis by promoting mitochondrial superoxide generation independent of Gli inhibition, and highlights the therapeutic potential of mitochondrial ROS-mediated anticancer drugs in MMe

Mutational Analysis of Hedgehog Signaling Pathway Genes in Human Malignant Mesothelioma

Background The Hedgehog (HH) signaling pathway is critical for embryonic development and adult homeostasis. Recent studies have identified regulatory roles for this pathway in certain cancers with mutations in the HH pathway genes. The extent to which mutations of the HH pathway genes are involved in the pathogenesis of malignant mesothelioma (MMe) is unknown. Methodology/Principal Findings Real-time PCR analysis of HH pathway genes PTCH1, GLI1 and GLI2 were performed on 7 human MMe cell lines. Exon sequencing of 13 HH pathway genes was also performed in cell lines and human MMe tumors. In silico programs were used to predict the likelihood that an amino-acid substitution would have a functional effect. GLI1, GLI2 and PTCH1 were highly expressed in MMe cells, indicative of active HH signaling. PTCH1, SMO and SUFU mutations were found in 2 of 11 MMe cell lines examined. A non-synonymous missense SUFU mutation (p.T411M) was identified in LO68 cells. In silico characterization of the SUFU mutant suggested that the p.T411M mutation might alter protein function. However, we were unable to demonstrate any functional effect of this mutation on Gli activity. Deletion of exons of the PTCH1 gene was found in JU77 cells, resulting in loss of one of two extracellular loops implicated in HH ligand binding and the intracellular C-terminal domain. A 3-bp insertion (69_70insCTG) in SMO, predicting an additional leucine residue in the signal peptide segment of SMO protein was also identified in LO68 cells and a MMe tumour. Conclusions/Significance We identified the first novel mutations in PTCH1, SUFU and SMO associated with MMe. Although HH pathway mutations are relatively rare in MMe, these data suggest a possible role for dysfunctional HH pathway in the pathogenesis of a subgroup of MMe and help rationalize the exploration of HH pathway inhibitors for MMe therapy

Large-scale collective motion of RFGC galaxies

We processed the data about radial velocities and HI linewidths for 1678 flat edge-on spirals from the Revised Flat Galaxy Catalogue. We obtained the parameters of the multipole components of large-scale velocity field of collective non-Hubble galaxy motion as well as the parameters of the generalized Tully-Fisher relationship in the "HI line width - linear diameter" version. All the calculations were performed independently in the framework of three models, where the multipole decomposition of the galaxy velocity field was limited to a dipole, quadrupole and octopole terms respectively. We showed that both the quadrupole and the octopole components are statistically significant. On the basis of the compiled list of peculiar velocities of 1623 galaxies we obtained the estimations of cosmological parameters Omega_m and sigma_8. This estimation is obtained in both graphical form and as a constraint of the value S_8=sigma_8(Omega_m/0.3)^0.35 = 0.91 +/- 0.05.Comment: Accepted for publication in Astrophysics and Space Scienc

Colliders and Cosmology

Dark matter in variations of constrained minimal supersymmetric standard models will be discussed. Particular attention will be given to the comparison between accelerator and direct detection constraints.Comment: Submitted for the SUSY07 proceedings, 15 pages, LaTex, 26 eps figure

Learning to learn: improving attainment, closing the gap at Key Stage 3

In 2010, a comprehensive secondary school in the south of England implemented a whole-school approach to ‘learning to learn’ (L2L). Drawing on a range of evidence-based practices, a team of teachers worked collaboratively to design and deliver a taught L2L curriculum to all students throughout Key Stage 3. In total, the first cohort of students (n = 118) received more than 400 taught lessons throughout years 7–9. The impact of L2L on student attainment at Sea View was evaluated over those 3 years, using the pre-L2L cohort as a matched control group (n = 148). By the end of year 9, a significantly higher proportion of L2L students were either hitting or exceeding their target grades, compared with the control group. There was also a significant closing of the attainment gap between students eligible for the pupil premium and their peers (2%, vs. 25% in the control group). Key features of the L2L approach at Sea View are considered in terms of similarities and differences with other L2L approaches. Conclusions are drawn that the success of this approach lies in the combination of multiple effective practices. Recommendations for further research and development of the field are proposed

Track A Basic Science

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/138319/1/jia218438.pd

Global, regional, and national incidence, prevalence, and years lived with disability for 310 diseases and injuries, 1990�2015: a systematic analysis for the Global Burden of Disease Study 2015

Background Non-fatal outcomes of disease and injury increasingly detract from the ability of the world's population to live in full health, a trend largely attributable to an epidemiological transition in many countries from causes affecting children, to non-communicable diseases (NCDs) more common in adults. For the Global Burden of Diseases, Injuries, and Risk Factors Study 2015 (GBD 2015), we estimated the incidence, prevalence, and years lived with disability for diseases and injuries at the global, regional, and national scale over the period of 1990 to 2015. Methods We estimated incidence and prevalence by age, sex, cause, year, and geography with a wide range of updated and standardised analytical procedures. Improvements from GBD 2013 included the addition of new data sources, updates to literature reviews for 85 causes, and the identification and inclusion of additional studies published up to November, 2015, to expand the database used for estimation of non-fatal outcomes to 60�900 unique data sources. Prevalence and incidence by cause and sequelae were determined with DisMod-MR 2.1, an improved version of the DisMod-MR Bayesian meta-regression tool first developed for GBD 2010 and GBD 2013. For some causes, we used alternative modelling strategies where the complexity of the disease was not suited to DisMod-MR 2.1 or where incidence and prevalence needed to be determined from other data. For GBD 2015 we created a summary indicator that combines measures of income per capita, educational attainment, and fertility (the Socio-demographic Index SDI) and used it to compare observed patterns of health loss to the expected pattern for countries or locations with similar SDI scores. Findings We generated 9·3 billion estimates from the various combinations of prevalence, incidence, and YLDs for causes, sequelae, and impairments by age, sex, geography, and year. In 2015, two causes had acute incidences in excess of 1 billion: upper respiratory infections (17·2 billion, 95% uncertainty interval UI 15·4�19·2 billion) and diarrhoeal diseases (2·39 billion, 2·30�2·50 billion). Eight causes of chronic disease and injury each affected more than 10% of the world's population in 2015: permanent caries, tension-type headache, iron-deficiency anaemia, age-related and other hearing loss, migraine, genital herpes, refraction and accommodation disorders, and ascariasis. The impairment that affected the greatest number of people in 2015 was anaemia, with 2·36 billion (2·35�2·37 billion) individuals affected. The second and third leading impairments by number of individuals affected were hearing loss and vision loss, respectively. Between 2005 and 2015, there was little change in the leading causes of years lived with disability (YLDs) on a global basis. NCDs accounted for 18 of the leading 20 causes of age-standardised YLDs on a global scale. Where rates were decreasing, the rate of decrease for YLDs was slower than that of years of life lost (YLLs) for nearly every cause included in our analysis. For low SDI geographies, Group 1 causes typically accounted for 20�30% of total disability, largely attributable to nutritional deficiencies, malaria, neglected tropical diseases, HIV/AIDS, and tuberculosis. Lower back and neck pain was the leading global cause of disability in 2015 in most countries. The leading cause was sense organ disorders in 22 countries in Asia and Africa and one in central Latin America; diabetes in four countries in Oceania; HIV/AIDS in three southern sub-Saharan African countries; collective violence and legal intervention in two north African and Middle Eastern countries; iron-deficiency anaemia in Somalia and Venezuela; depression in Uganda; onchoceriasis in Liberia; and other neglected tropical diseases in the Democratic Republic of the Congo. Interpretation Ageing of the world's population is increasing the number of people living with sequelae of diseases and injuries. Shifts in the epidemiological profile driven by socioeconomic change also contribute to the continued increase in years lived with disability (YLDs) as well as the rate of increase in YLDs. Despite limitations imposed by gaps in data availability and the variable quality of the data available, the standardised and comprehensive approach of the GBD study provides opportunities to examine broad trends, compare those trends between countries or subnational geographies, benchmark against locations at similar stages of development, and gauge the strength or weakness of the estimates available. Funding Bill & Melinda Gates Foundation. © 2016 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY licens

TP 227b Induction of mesothelioma cell apoptosis by GANT61, a small molecule inhibitor of GLI transcription factors: Evidence for redox-driven cytotoxicity

Background: Although it has been shown that Gli transcription factors are the major intracellular target of the small molecule inhibitor GANT61, the underlying mechanism by which GANT61 inhibits cancer cell growth remains unclear. In this study, we aimed to elucidate the mechanisms that underlie the induction of mesothelioma cell apoptosis by GANT61. Method: Human mesothelioma cells were treated with GANT61. Levels of apoptosis and reactive oxygen species (ROS) were quantified by flow cytometry. Results: Our study showed that GANT61 induces growth inhibition and apoptosis in mesothelioma cells through the induction of oxidative stress. Exposure of mesothelioma cells to GANT61 resulted in the rapid production of ROS. Quenching of ROS by the antioxidants N-acetylcysteine and L-glutathione blocked the induction of apoptosis and depolarization of mitochondrial membrane potential by GANT61, indicating that ROS overproduction is the primary driver for the pro-apoptotic activity of GANT61. Although GANT61 could inhibit Gli transcriptional activity, the inhibition of Gli was not sufficient to initiate apoptosis in mesothelioma cells. Further study revealed that the ROS species generated was superoxide from the mitochondria, and that mitochondrial electron transport chain inhibition by rotenone significantly suppressed the pro-apoptotic effect of GANT61. Conclusion: Our study provides evidence that GANT61 is a potent anti-mesothelioma therapeutic agent, which induces apoptosis in mesothelioma cells through the generation of mitochondrial superoxide