Albiglutide and cardiovascular outcomes in patients with type 2 diabetes and cardiovascular disease (Harmony Outcomes): a double-blind, randomised placebo-controlled trial

Background: Glucagon-like peptide 1 receptor agonists differ in chemical structure, duration of action, and in their effects on clinical outcomes. The cardiovascular effects of once-weekly albiglutide in type 2 diabetes are unknown. We aimed to determine the safety and efficacy of albiglutide in preventing cardiovascular death, myocardial infarction, or stroke. Methods: We did a double-blind, randomised, placebo-controlled trial in 610 sites across 28 countries. We randomly assigned patients aged 40 years and older with type 2 diabetes and cardiovascular disease (at a 1:1 ratio) to groups that either received a subcutaneous injection of albiglutide (30–50 mg, based on glycaemic response and tolerability) or of a matched volume of placebo once a week, in addition to their standard care. Investigators used an interactive voice or web response system to obtain treatment assignment, and patients and all study investigators were masked to their treatment allocation. We hypothesised that albiglutide would be non-inferior to placebo for the primary outcome of the first occurrence of cardiovascular death, myocardial infarction, or stroke, which was assessed in the intention-to-treat population. If non-inferiority was confirmed by an upper limit of the 95% CI for a hazard ratio of less than 1·30, closed testing for superiority was prespecified. This study is registered with ClinicalTrials.gov, number NCT02465515. Findings: Patients were screened between July 1, 2015, and Nov 24, 2016. 10 793 patients were screened and 9463 participants were enrolled and randomly assigned to groups: 4731 patients were assigned to receive albiglutide and 4732 patients to receive placebo. On Nov 8, 2017, it was determined that 611 primary endpoints and a median follow-up of at least 1·5 years had accrued, and participants returned for a final visit and discontinuation from study treatment; the last patient visit was on March 12, 2018. These 9463 patients, the intention-to-treat population, were evaluated for a median duration of 1·6 years and were assessed for the primary outcome. The primary composite outcome occurred in 338 (7%) of 4731 patients at an incidence rate of 4·6 events per 100 person-years in the albiglutide group and in 428 (9%) of 4732 patients at an incidence rate of 5·9 events per 100 person-years in the placebo group (hazard ratio 0·78, 95% CI 0·68–0·90), which indicated that albiglutide was superior to placebo (p<0·0001 for non-inferiority; p=0·0006 for superiority). The incidence of acute pancreatitis (ten patients in the albiglutide group and seven patients in the placebo group), pancreatic cancer (six patients in the albiglutide group and five patients in the placebo group), medullary thyroid carcinoma (zero patients in both groups), and other serious adverse events did not differ between the two groups. There were three (<1%) deaths in the placebo group that were assessed by investigators, who were masked to study drug assignment, to be treatment-related and two (<1%) deaths in the albiglutide group. Interpretation: In patients with type 2 diabetes and cardiovascular disease, albiglutide was superior to placebo with respect to major adverse cardiovascular events. Evidence-based glucagon-like peptide 1 receptor agonists should therefore be considered as part of a comprehensive strategy to reduce the risk of cardiovascular events in patients with type 2 diabetes. Funding: GlaxoSmithKline

Inhibition of GSK-3 Ameliorates A? Pathology in an Adult-Onset Drosophila Model of Alzheimer's Disease

Aβ peptide accumulation is thought to be the primary event in the pathogenesis of Alzheimer's disease (AD), with downstream neurotoxic effects including the hyperphosphorylation of tau protein. Glycogen synthase kinase-3 (GSK-3) is increasingly implicated as playing a pivotal role in this amyloid cascade. We have developed an adult-onset Drosophila model of AD, using an inducible gene expression system to express Arctic mutant Aβ42 specifically in adult neurons, to avoid developmental effects. Aβ42 accumulated with age in these flies and they displayed increased mortality together with progressive neuronal dysfunction, but in the apparent absence of neuronal loss. This fly model can thus be used to examine the role of events during adulthood and early AD aetiology. Expression of Aβ42 in adult neurons increased GSK-3 activity, and inhibition of GSK-3 (either genetically or pharmacologically by lithium treatment) rescued Aβ42 toxicity. Aβ42 pathogenesis was also reduced by removal of endogenous fly tau; but, within the limits of detection of available methods, tau phosphorylation did not appear to be altered in flies expressing Aβ42. The GSK-3–mediated effects on Aβ42 toxicity appear to be at least in part mediated by tau-independent mechanisms, because the protective effect of lithium alone was greater than that of the removal of tau alone. Finally, Aβ42 levels were reduced upon GSK-3 inhibition, pointing to a direct role of GSK-3 in the regulation of Aβ42 peptide level, in the absence of APP processing. Our study points to the need both to identify the mechanisms by which GSK-3 modulates Aβ42 levels in the fly and to determine if similar mechanisms are present in mammals, and it supports the potential therapeutic use of GSK-3 inhibitors in AD

Rbf/E2F1 control growth and endoreplication via steroid-independent Ecdysone Receptor signalling in Drosophila prostate-like secondary cells.

In prostate cancer, loss of the tumour suppressor gene, Retinoblastoma (Rb), and consequent activation of transcription factor E2F1 typically occurs at a late-stage of tumour progression. It appears to regulate a switch to an androgen-independent form of cancer, castration-resistant prostate cancer (CRPC), which frequently still requires androgen receptor (AR) signalling. We have previously shown that upon mating, binucleate secondary cells (SCs) of the Drosophila melanogaster male accessory gland (AG), which share some similarities with prostate epithelial cells, switch their growth regulation from a steroid-dependent to a steroid-independent form of Ecdysone Receptor (EcR) control. This physiological change induces genome endoreplication and allows SCs to rapidly replenish their secretory compartments, even when ecdysone levels are low because the male has not previously been exposed to females. Here, we test whether the Drosophila Rb homologue, Rbf, and E2F1 regulate this switch. Surprisingly, we find that excess Rbf activity reversibly suppresses binucleation in adult SCs. We also demonstrate that Rbf, E2F1 and the cell cycle regulators, Cyclin D (CycD) and Cyclin E (CycE), are key regulators of mating-dependent SC endoreplication, as well as SC growth in both virgin and mated males. Importantly, we show that the CycD/Rbf/E2F1 axis requires the EcR, but not ecdysone, to trigger CycE-dependent endoreplication and endoreplication-associated growth in SCs, mirroring changes seen in CRPC. Furthermore, Bone Morphogenetic Protein (BMP) signalling, mediated by the BMP ligand Decapentaplegic (Dpp), intersects with CycD/Rbf/E2F1 signalling to drive endoreplication in these fly cells. Overall, our work reveals a signalling switch, which permits rapid growth of SCs and increased secretion after mating, independently of previous exposure to females. The changes observed share mechanistic parallels with the pathological switch to hormone-independent AR signalling seen in CRPC, suggesting that the latter may reflect the dysregulation of a currently unidentified physiological process

Watching me, watching you: Privacy attitudes and reactions to identity card implementation scenarios in the United Kingdom

In the United Kingdom, government proposals for the introduction of an identity (ID) card have raised considerable privacy concerns. In the present research, opinions and attitudes about different ways of implementing ID cards are examined using an experimental methodology. Specifically, the level and type of compulsion and application process, and the use of a centralised database or trusted third party to hold personal information, are compared for attitudes towards ID cards. Moreover, the impact of implementation scenarios on people with different privacy concern profiles is examined. The results show that an implementation that combines high compulsion with a centralised database (the approach currently favoured by the UK Government) lead to the greatest negative shift in attitudes towards ID cards. Implementations proposed by others (e.g. the London School of Economics) show significantly less negative shift in attitudes. People’s pre-existing privacy concerns also influence their evaluation of the different implementation scenarios

DCGs are released from SCs during mating, activating BMP signalling.

<p><b>A, B</b>. SCs from 6-day-old esgF/O^ts virgin males (A) and from males immediately after mating (B) were fixed and stained with an anti-pMad antibody (red) and DAPI (blue), revealing that the proportion of SCs with detectable nuclear pMad is higher in mated animals. <b>C, D.</b> Immediately after mating, living SCs (D) have less GFP-GPI-labelled DCGs than virgins (C). Image shows a single z-plane of gland stained with Lysotracker Red; not all compartments are in the focal plane. Note that the largest MVBL in (C; arrowhead) contains GFP, probably because of fusion between a DCG compartment and the MVBL [<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1006366#pgen.1006366.ref017" target="_blank">17</a>]. <b>E</b>. Graph shows proportion of SCs with nuclear pMad in 6-day-old virgin, and mated males (dissected 8 min into mating [Mid] and immediately after mating), and mated males expressing <i>dpp</i>-RNAi and Dad in SCs from eclosion onwards using the <i>w; esg-GAL4 tub-GAL80</i>^<i>ts</i> <i>UAS-FLP; UAS-GFP</i>_<i>nls</i> <i>actin>FRT>CD2>FRT>GAL4</i> driver. <b>F</b>. Graph shows number of GFP-GPI-positive DCG compartments in 6-day-old virgin and mated males (using the <i>w; spi-GAL4 tub-GAL80</i>^<i>ts</i> <i>UAS-GFP-GPI</i> driver line; Double is twice mated in 2 h), and at different times after single mating in control SCs. Compartments were also counted in SCs expressing <i>Snap24</i> RNAi post-eclosion in virgins and immediately after mating. Labelled compartments were counted using a complete z-series for each cell. Genotypes for images are: <i>w; esg-GAL4 tub-GAL80</i>^<i>ts</i> <i>UAS-FLP/+; UAS-GFP</i>_<i>nls</i> <i>actin>FRT>CD2>FRT>GAL4/+</i> (A, B); <i>w; spi-GAL4 tub-GAL80</i>^<i>ts</i> <i>UAS-GFP-GPI/CyO</i> (C, D).***P<0.001, Kruskal-Wallis test, n>15. Scale bar for A-B is 20 μm and C-D is 10 μm.</p

Autocrine Dpp regulates SC growth, SV number, endolysosomal trafficking and exosome secretion.

<p><b>A</b>. Expression of <i>dpp</i>-RNAi during the first six days of adulthood using the esgF/O^ts driver reduces the size of SCs and their nuclei (green arrows) relative to MCs (red arrows). <b>B</b>. Relative SC:MC nuclear size for SCs expressing RNAis targeting <i>dpp</i> and <i>gbb</i>, or GFP-tagged Dpp and Gbb, revealing specific effects of Dpp on growth. <b>C</b>. <i>dpp</i>^<i>blk</i>-GAL4 drives expression of a UAS-coupled nuclear GFP exclusively in SCs of the AG. <b>D, E.</b> Mosaic expression of <i>dpp</i>-RNAi or Dpp-GFP in a subset of SCs has a stronger effect on nuclear growth in expressing cells (on–green arrows) than in non-expressing (off–red arrows; white dashed circle) SCs, although <i>dpp</i> knockdown also reduces growth in the latter. <b>F-K.</b> Co-expression of <i>dpp</i>-RNAi with CD63-GFP using the <i>dsx</i>-GAL4 driver (G) reduces non-acidic SV number (eg., marked by arrowhead) and increases GFP fluorescence in largest MVBL (arrow; stained with Lysotracker Red) compared to controls (F); the statistical analysis of these changes for two independent RNAis is shown in H and I respectively. Knockdown of <i>dpp</i> either results in a small increase in the size of the largest MVBL or no significant size change (J), and reduces exosome secretion (K). Confocal images are from fixed glands (A, C, D) stained with DAPI (blue) and for Fas3 (yellow) or from living glands (F, G). Genotypes for images are: <i>w; esg-GAL4 tub-GAL80</i>^<i>ts</i> <i>UAS-FLP; UAS-GFP</i>_<i>nls</i> <i>actin>FRT>CD2>FRT>GAL4/P[TRiP</i>.<i>HMS00011]attP2</i> (A and mosaic in D; the esgF/O^ts driver was also used to generate data in E); <i>w; P[w</i>^<i>+</i> <i>UAS-GFP</i>_<i>nls</i><i>]; P[w</i>^<i>+</i> <i>dpp</i>^<i>blk</i><i>-GAL4]</i> (C); <i>w; UAS-CD63-GFP tub-GAL80</i>^<i>ts</i><i>; dsx-GAL4</i> combined with no other transgene (F) or <i>P[TRiP</i>.<i>HMS00011]attP2</i> (III) (G). ***P<0.001, Kruskal-Wallis test, n = 10. Scale bar for A, D is 20 μm, F, G, 10 μm, and for C, 50 μm.</p

Model to explain autocrine regulation of DCG replenishment by BMP signalling in SCs.

<p>Schematics of a single SC immediately before and after mating. (1) In virgin males, Dpp is trafficked to and stored in DCGs. Sporadic release of these DCGs activates BMP signalling, and sustains a basal level of growth, DCG biogenesis and exosome secretion. (2) During mating, about 4 mature DCGs are released (3), resulting in an increase in BMP signalling (4), primarily via an autocrine mechanism and probably in pulses. This stimulates growth, but also increases biosynthesis of new DCG compartments (5; solid arrow), ensuring that the total number of DCGs is fully replenished within 24 h. Dashed arrows highlight other parts of the secretory/endolysosomal system that might be affected by altered BMP signalling. Previous data (Corrigan et al., 2014) and data presented here suggest that long-term elevated BMP signalling enhances endolysosomal trafficking.</p