RALA-mediated delivery of FKBPL nucleic acid therapeutics

Aims: RALA is a novel 30 mer bioinspired amphipathic peptide that is showing promise for gene delivery. Here, we used RALA to deliver the FK506-binding protein like – FKBPL gene (pFKBPL) – a novel member of the immunophilin protein family. FKBPL is a secreted protein, with overexpression shown to inhibit angiogenesis, tumor growth and stemness, through a variety of intra- and extracellular signaling mechanisms. We also elucidated proangiogenic activity and stemness after utilizing RALA to deliver siRNA (siFKBPL). Materials & methods: The RALA/pFKBPL and RALA/siFKBPL nanoparticles were characterized in terms of size, charge, stability and toxicity. Overexpression and knockdown of FKBPL was assessed in vitro and in vivo. Results: RALA delivered both pFKBPL and siFKBPL with less cytotoxicity than commercially available counterparts. In vivo, RALA/pFKBPL delivery retarded tumor growth, and prolonged survival with an associated decrease in angiogenesis, while RALA/siFKBPL had no effect on tumor growth rate or survival, but resulted in an increase in angiogenesis and stemness. Conclusion: RALA is an effective delivery system for both FKBPL DNA and RNAi and highlights an alternative therapeutic approach to harnessing FKBPL's antiangiogenic and antistemness activity.

Salt-inducible kinase 2 regulates mitotic progression and transcription in prostate cancer

UNLABELLED: Salt-inducible kinase 2 (SIK2) is a multifunctional kinase of the AMPK family that plays a role in CREB1-mediated gene transcription and was recently reported to have therapeutic potential in ovarian cancer. The expression of this kinase was investigated in prostate cancer clinical specimens. Interestingly, auto-antibodies against SIK2 were increased in the plasma of patients with aggressive disease. Examination of SIK2 in prostate cancer cells found that it functions both as a positive regulator of cell-cycle progression and a negative regulator of CREB1 activity. Knockdown of SIK2 inhibited cell growth, delayed cell-cycle progression, induced cell death, and enhanced CREB1 activity. Expression of a kinase-dead mutant of SIK2 also inhibited cell growth, induced cell death, and enhanced CREB1 activity. Treatment with a small-molecule SIK2 inhibitor (ARN-3236), currently in preclinical development, also led to enhanced CREB1 activity in a dose- and time-dependent manner. Because CREB1 is a transcription factor and proto-oncogene, it was posited that the effects of SIK2 on cell proliferation and viability might be mediated by changes in gene expression. To test this, gene expression array profiling was performed and while SIK2 knockdown or overexpression of the kinase-dead mutant affected established CREB1 target genes; the overlap with transcripts regulated by forskolin (FSK), the adenylate cyclase/CREB1 pathway activator, was incomplete.

IMPLICATIONS: This study demonstrates that targeting SIK2 genetically or therapeutically will have pleiotropic effects on cell-cycle progression and transcription factor activation, which should be accounted for when characterizing SIK2 inhibitors.

HES6 drives a critical AR transcriptional programme to induce castration-resistant prostate cancer through activation of an E2F1-mediated cell cycle network

Castrate-resistant prostate cancer (CRPC) is poorly characterized and heterogeneous and while the androgen receptor (AR) is of singular importance, other factors such as c-Myc and the E2F family also play a role in later stage disease. HES6 is a transcription co-factor associated with stem cell characteristics in neural tissue. Here we show that HES6 is up-regulated in aggressive human prostate cancer and drives castration-resistant tumour growth in the absence of ligand binding by enhancing the transcriptional activity of the AR, which is preferentially directed to a regulatory network enriched for transcription factors such as E2F1. In the clinical setting, we have uncovered a HES6-associated signature that predicts poor outcome in prostate cancer, which can be pharmacologically targeted by inhibition of PLK1 with restoration of sensitivity to castration. We have therefore shown for the first time the critical role of HES6 in the development of CRPC and identified its potential in patient-specific therapeutic strategies.

Improved Efficiency for Partial Oxidation of Methane by Controlled Copper Deposition on Surface-Modified ZSM-5

The mono(μ-oxo) dicopper cores present in the pores of Cu-ZSM-5 are active for the partial oxidation of methane to methanol. However, copper on the external surface reduces the ratio of active, selective sites to unselective sites. More efficient catalysts are obtained by controlling the copper deposition during synthesis. Herein, the external exchange sites of ZSM-5 samples were passivated by bis(trimethylsilyl) trifluoroacetamide (BSTFA) followed by calcination, promoting selective deposition of intraporous copper during aqueous copper ion exchange. At an optimum level of 1–2 wt % SiO2, IR studies showed a 64 % relative reduction in external copper species and temperature-programmed oxidation analysis showed an associated increase in the formation of methanol compared with unmodified Cu-ZSM-5 samples. It is, therefore, reported that the modified zeolites contained a significantly higher proportion of active, selective copper species than their unmodified counterparts with activity for partial methane oxidation to methanol.

Complementarity’s monopoly on justice in Uganda: the International Criminal Court, victims and Thomas Kwoyelo

Complementarity has been extolled as the pioneering way for the International Criminal Court (ICC) to navigate the difficulties of state sovereignty when investigating and prosecuting international crimes. Victims have often been held up to justify and legitimise the work of the ICC and states complementing the Court through domestic processes. This article examines how Uganda has developed its laws, legal procedure, and accountability for international crimes over the past decade. This has culminated in the trial of Thomas Kwoyelo, which after five years of proceedings, has yet to move to the trial phase, due to the issue of an amnesty. While there has been a profusion of provisions to allow victims to participate, avail of protection measures and reparations, in practice very little has changed for them. This article highlights the dangers of complementarity being the sole solution to protracted conflicts, in particular the realisation of victims’ rights.

In the Shadows of the Old Town: Thomas Anna and 19th-century Glasgow

In 1858, a volume entitled Midnight Scenes and Social Photographs – being sketches of life in the streets, wynds and dens of the city of Glasgow was published under the pseudonym of ‘Shadow’ by Alexander Brown, a Glaswegian flâneur and reformer. Its frontispiece is an etching which depicts a theatre-like proscenium arch whose curtains have been withdrawn to reveal to the audience all the poverty, destitution and disorder that one was likely to find after dark in the insalubrious quarters of the city. At the extreme left-hand side, partly obscured by the curtain a silhouetted figure stands behind an unwieldy camera perched on a tripod. Distinctly unaffected by the mêlée, an arm is calmly raised and a finger precisely arched in the moment before the shutter is clicked and the scene committed to record. The volume, however, relies exclusively on textual descriptions to evoke the underside of the city and contains no photographs at all. Instead, the use of the word photograph in the title can be understood as a metaphor for detached scientific objectivity, a quality much celebrated by nineteenth-century reformers and investigators of social ills. As it happened, a decade after Shadow disappeared into the labyrinthine back-lands of Old Town Glasgow, he was followed there by a real photographer. In 1868, Thomas Annan was commissioned by the City Improvements Trust to take photographs of the Old Town in its last moments of existence before it was pulled down under a series of legislative acts. But perhaps paradoxically, given Shadow’s faith in the analytical properties of photography, Annan’s work seems to refute much of the material contained in Midnight Scenes and other similar tracts. Instead of the dens, shebeens, labyrinths and rowdy crowds described by Shadow, Annan’s depictions of the Old Town convey a static, calm environment, one which is often sparsely inhabited by a curious but apparently orderly population.

Taking account of the sensational tendencies of many reformists’ texts, this paper investigates the discrepancies between the two representations, focussing in particular on the constraints which operated on Annan during his commission. It argues that Annan’s compositions – which became very influential on other 19th century photographers of everyday life such as John Thomson or Jacob Riis – far from being dispassionate analytical works, emerged as a result of a matrix of factors which included: photographic and artistic precedents; Annan’s own predilections as a photographer; technological limitations; the nature of the commission from the City Improvements Trust and political climate in which it was given; the medieval urban fabric in which he had to operate; and, perhaps, most importantly, the identity of the Old Towns inhabitants themselves.

Gene regulatory mechanisms underpinning prostate cancer susceptibility

Molecular characterization of genome-wide association study (GWAS) loci can uncover key genes and biological mechanisms underpinning complex traits and diseases. Here we present deep, high-throughput characterization of gene regulatory mechanisms underlying prostate cancer risk loci. Our methodology integrates data from 295 prostate cancer chromatin immunoprecipitation and sequencing experiments with genotype and gene expression data from 602 prostate tumor samples. The analysis identifies new gene regulatory mechanisms affected by risk locus SNPs, including widespread disruption of ternary androgen receptor (AR)-FOXA1 and AR-HOXB13 complexes and competitive binding mechanisms. We identify 57 expression quantitative trait loci at 35 risk loci, which we validate through analysis of allele-specific expression. We further validate predicted regulatory SNPs and target genes in prostate cancer cell line models. Finally, our integrated analysis can be accessed through an interactive visualization tool. This analysis elucidates how genome sequence variation affects disease predisposition via gene regulatory mechanisms and identifies relevant genes for downstream biomarker and drug development.