• Experimental long-term diabetes mellitus alters the transcriptome and biomechanical properties of the rat urinary bladder

      Hindi, Emad A.; Williams, Craig J.; Zeef, Leo A. H.; Lopes, Filipa M.; Newman, Katie; Davey, Martha M. M.; Hodson, Nigel W.; Hilton, Emma N.; Huang, Jennifer L.; Price, Karen L.; et al. (Nature Publishing Group UK, 2021-07-30)
      Abstract: Diabetes mellitus (DM) is the leading cause of chronic kidney disease and diabetic nephropathy is widely studied. In contrast, the pathobiology of diabetic urinary bladder disease is less understood despite dysfunctional voiding being common in DM. We hypothesised that diabetic cystopathy has a characteristic molecular signature. We therefore studied bladders of hyperglycaemic and polyuric rats with streptozotocin (STZ)-induced DM. Sixteen weeks after induction of DM, as assessed by RNA arrays, wide-ranging changes of gene expression occurred in DM bladders over and above those induced in bladders of non-hyperglycaemic rats with sucrose-induced polyuria. The altered transcripts included those coding for extracellular matrix regulators and neural molecules. Changes in key genes deregulated in DM rat bladders were also detected in db/db mouse bladders. In DM rat bladders there was reduced birefringent collagen between detrusor muscle bundles, and atomic force microscopy showed a significant reduction in tissue stiffness; neither change was found in bladders of sucrose-treated rats. Thus, altered extracellular matrix with reduced tissue rigidity may contribute to voiding dysfunction in people with long-term DM. These results serve as an informative stepping stone towards understanding the complex pathobiology of diabetic cystopathy.
    • Ezh2 is essential for the generation of functional yolk sac derived erythro-myeloid progenitors

      Neo, Wen Hao; orcid: 0000-0002-6827-3027; email: wenhao.neo@cruk.manchester.ac.uk; Meng, Yiran; orcid: 0000-0002-9333-2383; Rodriguez-Meira, Alba; Fadlullah, Muhammad Z. H.; Booth, Christopher A. G.; orcid: 0000-0003-3841-6637; Azzoni, Emanuele; orcid: 0000-0002-4572-5692; Thongjuea, Supat; orcid: 0000-0002-9129-4694; de Bruijn, Marella F. T. R.; orcid: 0000-0002-4934-4125; Jacobsen, Sten Eirik W.; Mead, Adam J.; orcid: 0000-0001-8522-1002; email: adam.mead@imm.ox.ac.uk; et al. (Nature Publishing Group UK, 2021-12-02)
      Abstract: Yolk sac (YS) hematopoiesis is critical for the survival of the embryo and a major source of tissue-resident macrophages that persist into adulthood. Yet, the transcriptional and epigenetic regulation of YS hematopoiesis remains poorly characterized. Here we report that the epigenetic regulator Ezh2 is essential for YS hematopoiesis but dispensable for subsequent aorta–gonad–mesonephros (AGM) blood development. Loss of EZH2 activity in hemogenic endothelium (HE) leads to the generation of phenotypically intact but functionally deficient erythro-myeloid progenitors (EMPs), while the generation of primitive erythroid cells is not affected. EZH2 activity is critical for the generation of functional EMPs at the onset of the endothelial-to-hematopoietic transition but subsequently dispensable. We identify a lack of Wnt signaling downregulation as the primary reason for the production of non-functional EMPs. Together, our findings demonstrate a critical and stage-specific role of Ezh2 in modulating Wnt signaling during the generation of EMPs from YS HE.
    • FAK inhibition alone or in combination with adjuvant therapies reduces cancer stem cell activity

      Timbrell, Simon; Aglan, Hosam; Cramer, Angela; Foden, Phil; Weaver, David; Pachter, Jonathan; Kilgallon, Aoife; Clarke, Robert B.; Farnie, Gillian; email: gillian.farnie@ndorms.ox.ac.uk; Bundred, Nigel J.; orcid: 0000-0001-6007-056X; email: Bundredn@manchester.ac.uk (Nature Publishing Group UK, 2021-05-28)
      Abstract: Cancer stem-like cells (CSC) contribute to therapy resistance and recurrence. Focal adhesion kinase (FAK) has a role in CSC regulation. We determined the effect of FAK inhibition on breast CSC activity alone and in combination with adjuvant therapies. FAK inhibition reduced CSC activity and self-renewal across all molecular subtypes in primary human breast cancer samples. Combined FAK and paclitaxel reduced self-renewal in triple negative cell lines. An invasive breast cancer cohort confirmed high FAK expression correlated with increased risk of recurrence and reduced survival. Co-expression of FAK and CSC markers was associated with the poorest prognosis, identifying a high-risk patient population. Combined FAK and paclitaxel treatment reduced tumour size, Ki67, ex-vivo mammospheres and ALDH+ expression in two triple negative patient derived Xenograft (PDX) models. Combined treatment reduced tumour initiation in a limiting dilution re-implantation PDX model. Combined FAK inhibition with adjuvant therapy has the potential to improve breast cancer survival.
    • First observation of radiolytic bubble formation in unstirred nano-powder sludges and a consistent model thereof

      O’Leary, Mel; email: mel.oleary@manchester.ac.uk; Baidak, Aliaksandr; Barnes, Martyn; Donoclift, Thomas; Emerson, Christopher; Figueira, Catarina; Fox, Oliver; Kleppe, Annette; McCulloch, Aaron; Messer, Darryl; et al. (Nature Publishing Group UK, 2021-11-24)
      Abstract: Experiments involving the irradiation of water contained within magnesium hydroxide and alumina nanoparticle sludges were conducted and culminated in observations of an increased yield of molecular hydrogen when compared to the yield from the irradiation of bulk water. We show that there is a relationship linking this increased yield to the direct nanoscale ionization mechanism in the nanoparticles, indicating that electron emission from the nanoparticles drives new radiative pathways in the water. Because the chemical changes in these sludges are introduced by irradiation only, we have a genuinely unstirred system. This feature allows us to determine the diffusivity of the dissolved gas. Using the measured gas production rate, we have developed a method for modelling when hydrogen bubble formation will occur within the nanoparticle sludges. This model facilitates the determination of a consistent radiolytic consumption rate coinciding with the observations of bubble formation. Thus, we demonstrate a nanoscale radiation effect directly influencing the formation of molecular hydrogen.
    • First-passage times and normal tissue complication probabilities in the limit of large populations

      Hufton, Peter G.; Buckingham-Jeffery, Elizabeth; email: e.buckingham-jeffery@manchester.ac.uk; Galla, Tobias (Nature Publishing Group UK, 2020-05-29)
      Abstract: The time of a stochastic process first passing through a boundary is important to many diverse applications. However, we can rarely compute the analytical distribution of these first-passage times. We develop an approximation to the first and second moments of a general first-passage time problem in the limit of large, but finite, populations using Kramers–Moyal expansion techniques. We demonstrate these results by application to a stochastic birth-death model for a population of cells in order to develop several approximations to the normal tissue complication probability (NTCP): a problem arising in the radiation treatment of cancers. We specifically allow for interaction between cells, via a nonlinear logistic growth model, and our approximations capture the effects of intrinsic noise on NTCP. We consider examples of NTCP in both a simple model of normal cells and in a model of normal and damaged cells. Our analytical approximation of NTCP could help optimise radiotherapy planning, for example by estimating the probability of complication-free tumour under different treatment protocols.
    • Focused VHEE (very high energy electron) beams and dose delivery for radiotherapy applications

      Whitmore, L.; Mackay, R. I.; van Herk, M.; Jones, J. K.; Jones, R. M.; email: Roger.Jones@manchester.ac.uk (Nature Publishing Group UK, 2021-07-07)
      Abstract: This paper presents the first demonstration of deeply penetrating dose delivery using focused very high energy electron (VHEE) beams using quadrupole magnets in Monte Carlo simulations. We show that the focal point is readily modified by linearly changing the quadrupole magnet strength only. We also present a weighted sum of focused electron beams to form a spread-out electron peak (SOEP) over a target region. This has a significantly reduced entrance dose compared to a proton-based spread-out Bragg peak (SOBP). Very high energy electron (VHEE) beams are an exciting prospect in external beam radiotherapy. VHEEs are less sensitive to inhomogeneities than proton and photon beams, have a deep dose reach and could potentially be used to deliver FLASH radiotherapy. The dose distributions of unfocused VHEE produce high entrance and exit doses compared to other radiotherapy modalities unless focusing is employed, and in this case the entrance dose is considerably improved over existing radiations. We have investigated both symmetric and asymmetric focusing as well as focusing with a range of beam energies.
    • Gene editing enables rapid engineering of complex antibiotic assembly lines

      Thong, Wei Li; Zhang, Yingxin; Zhuo, Ying; Robins, Katherine J.; orcid: 0000-0001-5049-4246; Fyans, Joanna K.; Herbert, Abigail J.; Law, Brian J. C.; Micklefield, Jason; orcid: 0000-0001-8951-4873; email: jason.micklefield@manchester.ac.uk (Nature Publishing Group UK, 2021-11-25)
      Abstract: Re-engineering biosynthetic assembly lines, including nonribosomal peptide synthetases (NRPS) and related megasynthase enzymes, is a powerful route to new antibiotics and other bioactive natural products that are too complex for chemical synthesis. However, engineering megasynthases is very challenging using current methods. Here, we describe how CRISPR-Cas9 gene editing can be exploited to rapidly engineer one of the most complex megasynthase assembly lines in nature, the 2.0 MDa NRPS enzymes that deliver the lipopeptide antibiotic enduracidin. Gene editing was used to exchange subdomains within the NRPS, altering substrate selectivity, leading to ten new lipopeptide variants in good yields. In contrast, attempts to engineer the same NRPS using a conventional homologous recombination-mediated gene knockout and complementation approach resulted in only traces of new enduracidin variants. In addition to exchanging subdomains within the enduracidin NRPS, subdomains from a range of NRPS enzymes of diverse bacterial origins were also successfully utilized.
    • Gene expression signatures predict response to therapy with growth hormone

      Stevens, Adam; orcid: 0000-0002-1950-7325; Murray, Philip; De Leonibus, Chiara; Garner, Terence; Koledova, Ekaterina; Ambler, Geoffrey; Kapelari, Klaus; Binder, Gerhard; Maghnie, Mohamad; Zucchini, Stefano; et al. (Nature Publishing Group UK, 2021-05-27)
      Abstract: Recombinant human growth hormone (r-hGH) is used as a therapeutic agent for disorders of growth including growth hormone deficiency (GHD) and Turner syndrome (TS). Treatment is costly and current methods to model response are inexact. GHD (n = 71) and TS patients (n = 43) were recruited to study response to r-hGH over 5 years. Analysis was performed using 1219 genetic markers and baseline (pre-treatment) blood transcriptome. Random forest was used to determine predictive value of transcriptomic data associated with growth response. No genetic marker passed the stringency criteria for prediction. However, we identified an identical set of genes in both GHD and TS whose expression could be used to classify therapeutic response to r-hGH with a high accuracy (AUC > 0.9). Combining transcriptomic markers with clinical phenotype was shown to significantly reduce predictive error. This work could be translated into a single genomic test linked to a prediction algorithm to improve clinical management. Trial registration numbers: NCT00256126 and NCT00699855.
    • Giant magneto-birefringence effect and tuneable colouration of 2D crystal suspensions

      Ding, Baofu; orcid: 0000-0001-6646-7285; Kuang, Wenjun; orcid: 0000-0003-4309-365X; Pan, Yikun; Grigorieva, I. V.; orcid: 0000-0001-5991-7778; Geim, A. K.; orcid: 0000-0003-2861-8331; email: geim@manchester.ac.uk; Liu, Bilu; orcid: 0000-0002-7274-5752; email: bilu.liu@sz.tsinghua.edu.cn; Cheng, Hui-Ming; orcid: 0000-0002-5387-4241; email: hmcheng@sz.tsinghua.edu.cn (Nature Publishing Group UK, 2020-07-24)
      Abstract: One of the long-sought-after goals in light manipulation is tuning of transmitted interference colours. Previous approaches toward this goal include material chirality, strain and electric-field controls. Alternatively, colour control by magnetic field offers contactless, non-invasive and energy-free advantages but has remained elusive due to feeble magneto-birefringence in conventional transparent media. Here we demonstrate an anomalously large magneto-birefringence effect in transparent suspensions of magnetic two-dimensional crystals, which arises from a combination of a large Cotton-Mouton coefficient and relatively high magnetic saturation birefringence. The effect is orders of magnitude stronger than those previously demonstrated for transparent materials. The transmitted colours of the suspension can be continuously tuned over two-wavelength cycles by moderate magnetic fields below 0.8 T. The work opens a new avenue to tune transmitted colours, and can be further extended to other systems with artificially engineered magnetic birefringence.
    • Global predictions of primary soil salinization under changing climate in the 21st century

      Hassani, Amirhossein; orcid: 0000-0002-6470-0490; email: ahas@nilu.no; Azapagic, Adisa; orcid: 0000-0003-2380-918X; email: adisa.azapagic@manchester.ac.uk; Shokri, Nima; orcid: 0000-0001-6799-4888; email: nima.shokri@tuhh.de (Nature Publishing Group UK, 2021-11-18)
      Abstract: Soil salinization has become one of the major environmental and socioeconomic issues globally and this is expected to be exacerbated further with projected climatic change. Determining how climate change influences the dynamics of naturally-occurring soil salinization has scarcely been addressed due to highly complex processes influencing salinization. This paper sets out to address this long-standing challenge by developing data-driven models capable of predicting primary (naturally-occurring) soil salinity and its variations in the world’s drylands up to the year 2100 under changing climate. Analysis of the future predictions made here identifies the dryland areas of South America, southern and western Australia, Mexico, southwest United States, and South Africa as the salinization hotspots. Conversely, we project a decrease in the soil salinity of the drylands in the northwest United States, the Horn of Africa, Eastern Europe, Turkmenistan, and west Kazakhstan in response to climate change over the same period.
    • Graphene oxide integrated silicon photonics for detection of vapour phase volatile organic compounds

      Leo Tsui, H. C.; Alsalman, Osamah; Mao, Boyang; Alodhayb, Abdullah; Albrithen, Hamad; Knights, Andrew P.; Halsall, Matthew P.; Crowe, Iain F.; email: iain.crowe@manchester.ac.uk (Nature Publishing Group UK, 2020-06-12)
      Abstract: The optical response of a graphene oxide integrated silicon micro-ring resonator (GOMRR) to a range of vapour phase Volatile Organic Compounds (VOCs) is reported. The response of the GOMRR to all but one (hexane) of the VOCs tested is significantly higher than that of the uncoated (control) silicon MRR, for the same vapour flow rate. An iterative Finite Difference Eigenmode (FDE) simulation reveals that the sensitivity of the GO integrated device (in terms of RIU/nm) is enhanced by a factor of ~2, which is coupled with a lower limit of detection. Critically, the simulations reveal that the strength of the optical response is determined by molecular specific changes in the local refractive index probed by the evanescent field of the guided optical mode in the device. Analytical modelling of the experimental data, based on Hill-Langmuir adsorption characteristics, suggests that these changes in the local refractive index are determined by the degree of molecular cooperativity, which is enhanced for molecules with a polarity that is high, relative to their kinetic diameter. We believe this reflects a molecular dependent capillary condensation within the graphene oxide interlayers, which, when combined with highly sensitive optical detection, provides a potential route for discriminating between different vapour phase VOCs.
    • Heritability of haemodynamics in the ascending aorta

      McGurk, Kathryn A.; email: K.McGurk@imperial.ac.uk; Owen, Benjamin; Watson, William D.; Nethononda, Richard M.; Cordell, Heather J.; Farrall, Martin; Rider, Oliver J.; Watkins, Hugh; Revell, Alistair; Keavney, Bernard D.; email: Bernard.Keavney@manchester.ac.uk (Nature Publishing Group UK, 2020-09-01)
      Abstract: Blood flow in the vasculature can be characterised by dimensionless numbers commonly used to define the level of instabilities in the flow, for example the Reynolds number, Re. Haemodynamics play a key role in cardiovascular disease (CVD) progression. Genetic studies have identified mechanosensitive genes with causal roles in CVD. Given that CVD is highly heritable and abnormal blood flow may increase risk, we investigated the heritability of fluid metrics in the ascending aorta calculated using patient-specific data from cardiac magnetic resonance (CMR) imaging. 341 participants from 108 British Caucasian families were phenotyped by CMR and genotyped for 557,124 SNPs. Flow metrics were derived from the CMR images to provide some local information about blood flow in the ascending aorta, based on maximum values at systole at a single location, denoted max, and a ‘peak mean’ value averaged over the area of the cross section, denoted pm. Heritability was estimated using pedigree-based (QTDT) and SNP-based (GCTA-GREML) methods. Estimates of Reynolds number based on spatially averaged local flow during systole showed substantial heritability (hPed2=41%[P=0.001], hSNP2=39%[P=0.002]), while the estimated heritability for Reynolds number calculated using the absolute local maximum velocity was not statistically significant (12–13%; P>0.05). Heritability estimates of the geometric quantities alone; e.g. aortic diameter (hPed2=29%[P=0.009], hSNP2=30%[P=0.010]), were also substantially heritable, as described previously. These findings indicate the potential for the discovery of genetic factors influencing haemodynamic traits in large-scale genotyped and phenotyped cohorts where local spatial averaging is used, rather than instantaneous values. Future Mendelian randomisation studies of aortic haemodynamic estimates, which are swift to derive in a clinical setting, will allow for the investigation of causality of abnormal blood flow in CVD.
    • High-energy synchrotron X-ray tomography coupled with digital image correlation highlights likely failure points inside ITER toroidal field conductors

      Warr, Ryan; Jewell, Matthew C.; Mitchell, Neil; Rack, Alexander; Swanson, Jack; Tronza, Vladimir; Cernik, Robert; email: b.cernik@manchester.ac.uk (Nature Publishing Group UK, 2021-11-30)
      Abstract: Two sections of heat-treated (HT) and non-heat-treated (NHT) Cable-in-Conduit Conductor (CICC) of a design similar to the ITER tokomak have been imaged using very high energy X-ray tomography at the ESRF beamline ID19. The sample images were collected at four temperatures down to 77 K. These results showed a greater degree of movement, bundle distortion and touching strands in the NHT sample. The HT sample showed non-linear movements with temperature especially close to 77 K; increasing non-circularity of the superconducting fibre bundles towards the periphery of the CICC, and touching bundles throughout the CICC. The images have highlighted where future design might improve potential weakness, in particular at the outer perimeters of the conductor and the individual sub-cable, ‘petal’ wraps.
    • How do women experience a false-positive test result from breast screening? A systematic review and thematic synthesis of qualitative studies

      Long, Hannah; orcid: 0000-0001-7306-8987; email: hannah.long@manchester.ac.uk; Brooks, Joanna M.; Harvie, Michelle; orcid: 0000-0001-9761-3089; Maxwell, Anthony; orcid: 0000-0001-8344-4958; French, David P.; orcid: 0000-0002-7663-7804 (Nature Publishing Group UK, 2019-07-23)
      Abstract: Background: This is the first review to identify, appraise and synthesise women’s experiences of having a false-positive breast screening test result. Methods: We systematically searched eight databases for qualitative research reporting women’s experiences of receiving a false-positive screening test result. Two reviewers independently screened articles. Eight papers reporting seven studies were included. Study quality was appraised. Data were thematically synthesised. Results: Women passively attended screening in order to prove their perceived good health. Consequently, being recalled was unexpected, shocking and disempowering: women felt without options. They endured great uncertainty and stress and sought clarity about their health (e.g. by scrutinising the wording of recall letters and conversations with healthcare professionals). Their result was accompanied by relief and welcome feelings of certainty about their health, but some received unclear explanations of their result, contributing to lasting breast cancer-related worry and an ongoing need for further reassurance. Conclusion: The organisation of breast screening programmes may constrain choice for women: they became passive recipients. The way healthcare professionals verbally communicate results to women may contribute to lasting breast cancer-related worry. Women need more reassurance, emotional support and answers to their questions before and during screening assessment, and after receiving their result.
    • Impact failure in two silicates revealed by ultrafast, in situ, synchrotron X-ray microscopy

      Bourne, N. K.; email: neil.bourne@manchester.ac.uk; Mirihanage, W. U.; Olbinado, M. P.; Rack, A.; Rau, C. (Nature Publishing Group UK, 2020-06-25)
      Abstract: To travel safely behind screens that can protect us from stones and hail, we must understand the response of glass to impact. However, without a means to observe the mechanisms that fail different silicate architectures, engineering has relied on external sensors, post-impact examination and best-guess to glaze our vehicles. We have used single and multi-bunch, X-ray imaging to differentiate distinct phases of failure in two silicates. We identified distinct micromechanisms, operating in tandem and leading to failure in borosilicate glass and Z-cut quartz. A surface zone in the amorphous glass densifies before bulk fracture occurs and then fails the block, whilst in quartz, fast cracks, driven down cleavage planes, fails the bulk. Varying the rate at which ejecta escapes by using different indenter tip geometries controls the failed target’s bulk strength. This opens the way to more physically based constitutive descriptions for the glasses allowing design of safer, composite panels by controlling the impulses felt by protective screens.
    • In-situ nanospectroscopic imaging of plasmon-induced two-dimensional [4+4]-cycloaddition polymerization on Au(111)

      Shao, Feng; orcid: 0000-0003-3879-5884; email: feng.shao@manchester.ac.uk; Wang, Wei; Yang, Weimin; Yang, Zhilin; Zhang, Yao; orcid: 0000-0002-6524-0289; Lan, Jinggang; email: jinggang.lan@chem.uzh.ch; Dieter Schlüter, A.; Zenobi, Renato; orcid: 0000-0001-5211-4358; email: zenobi@org.chem.ethz.ch (Nature Publishing Group UK, 2021-07-27)
      Abstract: Plasmon-induced chemical reactions (PICRs) have recently become promising approaches for highly efficient light-chemical energy conversion. However, an in-depth understanding of their mechanisms at the nanoscale still remains challenging. Here, we present an in-situ investigation by tip-enhanced Raman spectroscopy (TERS) imaging of the plasmon-induced [4+4]-cycloaddition polymerization within anthracene-based monomer monolayers physisorbed on Au(111), and complement the experimental results with density functional theory (DFT) calculations. This two-dimensional (2D) polymerization can be flexibly triggered and manipulated by the hot carriers, and be monitored simultaneously by TERS in real time and space. TERS imaging provides direct evidence for covalent bond formation with ca. 3.7 nm spatial resolution under ambient conditions. Combined with DFT calculations, the TERS results demonstrate that the lateral polymerization on Au(111) occurs by a hot electron tunneling mechanism, and crosslinks form via a self-stimulating growth mechanism. We show that TERS is promising to be plasmon-induced nanolithography for organic 2D materials.
    • Information content best characterises the hemispheric selectivity of the inferior parietal lobe: a meta-analysis

      Gray, Oliver; email: oliver.gray@manchester.ac.uk; Fry, Lewis; Montaldi, Daniela (Nature Publishing Group UK, 2020-09-15)
      Abstract: Our understanding of the inferior parietal lobe (IPL) remains challenged by inconsistencies between neuroimaging and neuropsychological perspectives. To date, others assume that hemispheric specialisation of the IPL is linked with the type of processing; attention processing in the right hemisphere; memory retrieval and semantic judgement in the left hemisphere. Here, we provide compelling evidence associating the type of information being processed with the recruitment of each hemisphere’s IPL. In a meta-analysis, we classify 121 previous fMRI reports of IPL activity arising from episodic memory retrieval, according to the type of information that characterises each fMRI contrast. We demonstrate that the left IPL is more consistently associated with retrieval of the semantic (95% of eligible contrasts) than perceptual aspects of memory (83%). In contrast, the right IPL is more consistently associated with the retrieval of perceptual (97%), than semantic aspects of memory (43%). This work revises assumptions of how the IPL contributes to healthy cognition and has major implications for IPL-related neuropsychological deficits.
    • Insulin protects acinar cells during pancreatitis by preserving glycolytic ATP supply to calcium pumps

      Bruce, Jason I. E.; orcid: 0000-0002-4503-1981; email: jason.bruce@manchester.ac.uk; Sánchez-Alvarez, Rosa; Sans, Maria Dolors; orcid: 0000-0002-9271-2106; Sugden, Sarah A.; Qi, Nathan; James, Andrew D.; orcid: 0000-0002-2432-5948; Williams, John A. (Nature Publishing Group UK, 2021-07-19)
      Abstract: Acute pancreatitis (AP) is serious inflammatory disease of the pancreas. Accumulating evidence links diabetes with severity of AP, suggesting that endogenous insulin may be protective. We investigated this putative protective effect of insulin during cellular and in vivo models of AP in diabetic mice (Ins2Akita) and Pancreatic Acinar cell-specific Conditional Insulin Receptor Knock Out mice (PACIRKO). Caerulein and palmitoleic acid (POA)/ethanol-induced pancreatitis was more severe in both Ins2Akita and PACIRKO vs control mice, suggesting that endogenous insulin directly protects acinar cells in vivo. In isolated pancreatic acinar cells, insulin induced Akt-mediated phosphorylation of 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 2 (PFKFB2) which upregulated glycolysis thereby preventing POA-induced ATP depletion, inhibition of the ATP-dependent plasma membrane Ca2+ ATPase (PMCA) and cytotoxic Ca2+ overload. These data provide the first mechanistic link between diabetes and severity of AP and suggest that phosphorylation of PFKFB2 may represent a potential therapeutic strategy for treatment of AP.
    • Ipomoeassin-F disrupts multiple aspects of secretory protein biogenesis

      Roboti, Peristera; email: peristera.roboti@manchester.ac.uk; O’Keefe, Sarah; Duah, Kwabena B.; Shi, Wei Q.; High, Stephen; email: stephen.high@manchester.ac.uk (Nature Publishing Group UK, 2021-06-02)
      Abstract: The Sec61 complex translocates nascent polypeptides into and across the membrane of the endoplasmic reticulum (ER), providing access to the secretory pathway. In this study, we show that Ipomoeassin-F (Ipom-F), a selective inhibitor of protein entry into the ER lumen, blocks the in vitro translocation of certain secretory proteins and ER lumenal folding factors whilst barely affecting others such as albumin. The effects of Ipom-F on protein secretion from HepG2 cells are twofold: reduced ER translocation combined, in some cases, with defective ER lumenal folding. This latter issue is most likely a consequence of Ipom-F preventing the cell from replenishing its ER lumenal chaperones. Ipom-F treatment results in two cellular stress responses: firstly, an upregulation of stress-inducible cytosolic chaperones, Hsp70 and Hsp90; secondly, an atypical unfolded protein response (UPR) linked to the Ipom-F-mediated perturbation of ER function. Hence, although levels of spliced XBP1 and CHOP mRNA and ATF4 protein increase with Ipom-F, the accompanying increase in the levels of ER lumenal BiP and GRP94 seen with tunicamycin are not observed. In short, although Ipom-F reduces the biosynthetic load of newly synthesised secretory proteins entering the ER lumen, its effects on the UPR preclude the cell restoring ER homeostasis.
    • Large cell neuroendocrine lung carcinoma: consensus statement from The British Thoracic Oncology Group and the Association of Pulmonary Pathologists

      Lindsay, Colin R.; email: colin.lindsay@manchester.ac.uk; Shaw, Emily C.; Moore, David A.; Rassl, Doris; Jamal-Hanjani, Mariam; Steele, Nicola; Naheed, Salma; Dick, Craig; Taylor, Fiona; Adderley, Helen; et al. (Nature Publishing Group UK, 2021-09-06)
      Abstract: Over the past 10 years, lung cancer clinical and translational research has been characterised by exponential progress, exemplified by the introduction of molecularly targeted therapies, immunotherapy and chemo-immunotherapy combinations to stage III and IV non-small cell lung cancer. Along with squamous and small cell lung cancers, large cell neuroendocrine carcinoma (LCNEC) now represents an area of unmet need, particularly hampered by the lack of an encompassing pathological definition that can facilitate real-world and clinical trial progress. The steps we have proposed in this article represent an iterative and rational path forward towards clinical breakthroughs that can be modelled on success in other lung cancer pathologies.