CMTM6 maintains the expression of PD-L1 and regulates anti-tumour immunity

Cancer cells exploit the expression of the programmed death-1 (PD-1) ligand 1 (PD-L1) to subvert T-cell-mediated immunosurveillance. The success of therapies that disrupt PD-L1-mediated tumour tolerance has highlighted the need to understand the molecular regulation of PD-L1 expression1. Here we identify the uncharacterized protein CMTM6 as a critical regulator of PD-L1 in a broad range of cancer cells, by using a genome-wide CRISPR–Cas9 screen. CMTM6 is a ubiquitously expressed protein that binds PD-L1 and maintains its cell surface expression. CMTM6 is not required for PD-L1 maturation but co-localizes with PD-L1 at the plasma membrane and in recycling endosomes, where it prevents PD-L1 from being targeted for lysosome-mediated degradation. Using a quantitative approach to profile the entire plasma membrane proteome, we find that CMTM6 displays specificity for PD-L1. Notably, CMTM6 depletion decreases PD-L1 without compromising cell surface expression of MHC class I. CMTM6 depletion, via the reduction of PD-L1, significantly alleviates the suppression of tumour-specific T cell activity in vitro and in vivo. These findings provide insights into the biology of PD-L1 regulation, identify a previously unrecognized master regulator of this critical immune checkpoint and highlight a potential therapeutic target to overcome immune evasion by tumour cells.M.L.B. is supported by a Cancer Research UK Clinician Scientist Fellowship (C53779/A20097), Addenbrooke’s Charitable Trust award and NIHR fellowship. P.J.L. is supported by a Wellcome Trust PRF (101835/Z/13/Z) and work in the Lehner laboratory is supported by NHSBT, NIHR Cambridge BRC, a Wellcome Trust Strategic Award to CIMR, and the Addenbrooke’s Charitable Trust M.A.D. is supported by a Senior Leukaemia Foundation Australia Fellowship and work in the Dawson laboratory is supported by the NHMRC (Grants 1085015, 1106444 and 1106447) Cancer Council Victoria and Leukaemia Foundation Australia. Cancer Research UK (C53779/A20097

Older people's experiences of changed medication appearance due to generic prescribing: a qualitative study

Research was undertaken in response to requests by older people in Rochdale Borough to investigate an issue of great concern to them namely prescription medications constantly changing appearance due to generic prescribing. When branded drugs are still in their ‘patent’ period, which is a number of years after they are introduced into the market, there is only one sole supplier. Expiration of this period allows other manufacturers to ‘mimic’ the branded product, at a fraction of the cost. It is an NHS directive for prescribers to use the generic drug name when a drug is prescribed unless there is a clinical justification for using the brand name. When people receive their tablet medicines from their pharmacist, the brand and so the appearance (colour, size, shape) can be vastly different to those dispensed following their previous prescription despite having the same active ingredient. This is often due to a lack of standardisation practice required amongst manufacturers. Drugs are made to British and European Pharmacopoeia standards but these do not specify colour, size and shape. Medicines are required to be of 'essential similarity' but that does not include appearance. Many pharmacists believe that standardisation, should not only include size, shape, colour but also packaging. Packaging changes from the same company on consecutive orders presents considerable challenge to pharmacists as well as patients.This patient experience study involved in-depth qualitative interviews with 32 people across Greater Manchester aged between 62 and 88 years of age taking three or more prescribed tablet medications. Findings revealed that these older people were challenged by the changes in their medications which impacted on their confidence and forced them to repeatedly double check their medicine regimes. The older people we interviewed felt they were only able to cope with the changes in medicine appearance due to their own diligence and because they had capacity to do so. Most were very concerned that as they became older and their capacity to self-manage medicines reduced, that they would be at risk of medication errors. They also believed there were many older people less capable than themselves who were already struggling to cope with medicine appearance changes. One participant said “You wouldn’t buy an orange cabbage would you?” to illustrate how disconcerting they found changes to their established medicines. This study builds on the team’s other Greater Manchester-wide research of the same topic which was a survey of 2000 older people and which painted a much more negative picture (Williamson et al 2009) [available from http://usir.salford.ac.uk/2989/] and many older people reported being stressed or confused with the changes in medication appearance. Study recommendations centre on future research and multiple agency work to:• Improve written information and information processes • Consider of the use of pictures on boxes of medications• Require manufacturers to notify pharmacists and explain changes and provide standardised packaging in terms of quantity (28 or 30 day packs)• Scope individual strategies - formal and informal - that people use to manage their medicines at home as a means of identifying risk • Identify training / education needs amongst primary care workers e.g. district nurses, pharmacists and GPs and partners such as Local Authorities in supporting older people with their medicines and identifying those most at risk e.g. vulnerable adults• Explore the frequency and nature of changes to individual’s medication appearance • Undertake economic analyses of generic prescribing practices including cost and quality of life implications• Explore the impact of changed appearance of medication with the wider older population including those who are less able to participate e.g. seldom heard or marginalised groups, those who are socially isolated and especially those at higher risk of negative effects such as those with reduced capacity to self manage medications• Devise a public information poster explaining 1) how large savings from generic prescribing are reinvested into patient services and 2) how when drugs come off patent, other manufacturers are free to produce them at lower cost, as long as they maintain efficacy 3) how pharmacists cannot order medicines from a specific manufacturer as it depends what wholesaler has in stock 4) the requirement to bulk some tablets up with excipients 5) the need for patients to talk to their pharmacis

The DNA methylome of human sperm is distinct from blood with little evidence for tissue-consistent obesity associations

Epidemiological research suggests that paternal obesity may increase the risk of fathering small for gestational age offspring. Studies in non-human mammals indicate that such associations could be mediated by DNA methylation changes in spermatozoa that influence offspring development in utero. Human obesity is associated with differential DNA methylation in peripheral blood. It is unclear, however, whether this differential DNA methylation is reflected in spermatozoa. We profiled genome-wide DNA methylation using the Illumina MethylationEPIC array in a cross-sectional study of matched human blood and sperm from lean (discovery n = 47; replication n = 21) and obese (n = 22) males to analyse tissue covariation of DNA methylation, and identify obesity-associated methylomic signatures. We found that DNA methylation signatures of human blood and spermatozoa are highly discordant, and methylation levels are correlated at only a minority of CpG sites (~1%). At the majority of these sites, DNA methylation appears to be influenced by genetic variation. Obesity-associated DNA methylation in blood was not generally reflected in spermatozoa, and obesity was not associated with altered covariation patterns or accelerated epigenetic ageing in the two tissues. However, one cross-tissue obesity-specific hypermethylated site (cg19357369; chr4:2429884; P = 8.95 × 10^{-8}; 2% DNA methylation difference) was identified, warranting replication and further investigation. When compared to a wide range of human somatic tissue samples (n = 5,917), spermatozoa displayed differential DNA methylation across pathways enriched in transcriptional regulation. Overall, human sperm displays a unique DNA methylation profile that is highly discordant to, and practically uncorrelated with, that of matched peripheral blood. We observed that obesity was only nominally associated with differential DNA methylation in sperm, and therefore suggest that spermatozoal DNA methylation is an unlikely mediator of intergenerational effects of metabolic traits

Photoecology of the Antarctic cyanobacterium Leptolyngbya sp. BC1307 brought to light through community analysis, comparative genomics and in vitro photophysiology

Cyanobacteria are important photoautotrophs in extreme environments such as the McMurdo Dry Valleys, Antarctica. Terrestrial Antarctic cyanobacteria experience constant darkness during the winter and constant light during the summer which influences the ability of these organisms to fix carbon over the course of an annual cycle. Here, we present a unique approach combining community structure, genomic and photophysiological analyses to understand adaptation to Antarctic light regimes in the cyanobacterium Leptolyngbya sp. BC1307. We show that Leptolyngbya sp. BC1307 belongs to a clade of cyanobacteria that inhabits near‐surface environments in the McMurdo Dry Valleys. Genomic analyses reveal that, unlike close relatives, Leptolyngbya sp. BC1307 lacks the genes necessary for production of the pigment phycoerythrin and is incapable of complimentary chromatic acclimation, while containing several genes responsible for known photoprotective pigments. Photophysiology experiments confirmed Leptolyngbya sp. BC1307 to be tolerant of short‐term exposure to high levels of photosynthetically active radiation, while sustained exposure reduced its capacity for photoprotection. As such, Leptolyngbya sp. BC1307 likely exploits low‐light microenvironments within cyanobacterial mats in the McMurdo Dry Valleys

Whole-genome sequencing of Salmonella Mississippi and Typhimurium Definitive Type 160, Australia and New Zealand

We used phylogenomic and risk factor data on isolates of Salmonella enterica serovars Mississippi and Typhimurium definitive type 160 (DT160) collected from human, animal, and environmental sources to elucidate their epidemiology and disease reservoirs in Australia and New Zealand. Sequence data suggested wild birds as a likely reservoir for DT160; animal and environmental sources varied more for Salmonella Mississippi than for Salmonella Typhimurium. Australia and New Zealand isolates sat in distinct clades for both serovars; the median single-nucleotide polymorphism distance for DT160 was 29 (range 8–66) and for Salmonella Mississippi, 619 (range 565–737). Phylogenomic data identified plausible sources of human infection from wildlife and environmental reservoirs and provided evidence supporting New Zealand–acquired DT160 in a group of travelers returning to Australia. Wider use of real-time whole-genome sequencing in new locations and for other serovars may identify sources and routes of transmission, thereby aiding prevention and control

How and why DNA barcodes underestimate the diversity of microbial eukaryotes

Background: Because many picoplanktonic eukaryotic species cannot currently be maintained in culture, direct sequencing of PCR-amplified 18S ribosomal gene DNA fragments from filtered sea-water has been successfully used to investigate the astounding diversity of these organisms. The recognition of many novel planktonic organisms is thus based solely on their 18S rDNA sequence. However, a species delimited by its 18S rDNA sequence might contain many cryptic species, which are highly differentiated in their protein coding sequences. Principal Findings: Here, we investigate the issue of species identification from one gene to the whole genome sequence. Using 52 whole genome DNA sequences, we estimated the global genetic divergence in protein coding genes between organisms from different lineages and compared this to their ribosomal gene sequence divergences. We show that this relationship between proteome divergence and 18S divergence is lineage dependant. Unicellular lineages have especially low 18S divergences relative to their protein sequence divergences, suggesting that 18S ribosomal genes are too conservative to assess planktonic eukaryotic diversity. We provide an explanation for this lineage dependency, which suggests that most species with large effective population sizes will show far less divergence in 18S than protein coding sequences. Conclusions: There is therefore a trade-off between using genes that are easy to amplify in all species, but which by their nature are highly conserved and underestimate the true number of species, and using genes that give a better description of the number of species, but which are more difficult to amplify. We have shown that this trade-off differs between unicellular and multicellular organisms as a likely consequence of differences in effective population sizes. We anticipate that biodiversity of microbial eukaryotic species is underestimated and that numerous ''cryptic species'' will become discernable with the future acquisition of genomic and metagenomic sequences

New mutations at the imprinted Gnas cluster show gene dosage effects of Gsα in postnatal growth and implicate XLαs in bone and fat metabolism, but not in suckling

The imprinted Gnas cluster is involved in obesity, energy metabolism, feeding behavior, and viability. Relative contribution of paternally expressed proteins XLαs, XLN1, and ALEX or a double dose of maternally expressed Gsα to phenotype has not been established. In this study, we have generated two new mutants (Ex1A-T-CON and Ex1A-T) at the Gnas cluster. Paternal inheritance of Ex1A-T-CON leads to loss of imprinting of Gsα, resulting in preweaning growth retardation followed by catch-up growth. Paternal inheritance of Ex1A-T leads to loss of imprinting of Gsα and loss of expression of XLαs and XLN1. These mice have severe preweaning growth retardation and incomplete catch-up growth. They are fully viable probably because suckling is unimpaired, unlike mutants in which the expression of all the known paternally expressed Gnasxl proteins (XLαs, XLN1 and ALEX) is compromised. We suggest that loss of ALEX is most likely responsible for the suckling defects previously observed. In adults, paternal inheritance of Ex1A-T results in an increased metabolic rate and reductions in fat mass, leptin, and bone mineral density attributable to loss of XLαs. This is, to our knowledge, the first report describing a role for XLαs in bone metabolism. We propose that XLαs is involved in the regulation of bone and adipocyte metabolism

A report on the health and social care listening event

The purpose of the Listening Event was to enable a wide range of people, including professionals working in statutory, voluntary and other organisations and members of the public, to ‘have a say’ about health and social care and what we as a University can do for and with these partners and the public. We particularly wanted to hear about key concerns of the University such as: • Strengthening community engagement and partnerships • Health and social care training we should be providing, for whom, and how this is delivered • Ideas relating to the University themes including media, use of space and buildings, human rights, social justice and security • Research topics we should be addressing However the main strength of the Listening Event approach is that topics for discussion are mostly led by participants who attend. On this occasion, the discussion topics were very much focused on the concerns of participants and lots of information and ideas were generated. The task now is for the event planning team to review the discussion notes and identify what can be addressed and how, in the short, medium and long term. This planning will be taking place over the Autumn in 2011, and any participants or readers of this report are more than welcome to get in touch to work with us or add their views. The purpose of this report is to record all discussion summaries for sharing amongst participants and others. It is important that participants especially get to read what others had said at the event. The report will lead to changes in University practices such as the content of some of our courses and new business ideas and relationships will also be explored. The event itself provided a useful means of public engagement that others may wish to adopt

Mobile phones are a viable option for surveying young Australian women: a comparison of two telephone survey methods

<p>Abstract</p> <p>Background</p> <p>Households with fixed-line telephones have decreased while mobile (cell) phone ownership has increased. We therefore sought to examine the feasibility of recruiting young women for a national health survey through random digit dialling mobile phones.</p> <p>Methods</p> <p>Two samples of women aged 18 to 39 years were surveyed by random digit dialling fixed and mobile numbers. We compared participation rates and responses to a questionnaire between women surveyed by each contact method.</p> <p>Results</p> <p>After dialling 5,390 fixed-lines and 3,697 mobile numbers, 140 and 128 women were recruited respectively. Among women contacted and found to be eligible, participation rates were 74% for fixed-lines and 88% for mobiles. Taking into account calls to numbers where eligibility was unknown (e.g. unanswered calls) the estimated response rates were 54% and 45% respectively. Of women contacted by fixed-line, 97% reported having a mobile while 61% of those contacted by mobile reported having a fixed-line at home. After adjusting for age, there were no significant differences between mobile-only and fixed-line responders with respect to education, residence, and various health behaviours; however compared to those with fixed-lines, mobile-only women were more likely to identify as Indigenous (OR 4.99, 95%CI 1.52-16.34) and less likely to live at home with their parents (OR 0.09, 95%CI 0.03-0.29).</p> <p>Conclusions</p> <p>Random digit dialling mobile phones to conduct a health survey in young Australian women is feasible, gives a comparable response rate and a more representative sample than dialling fixed-lines only. Telephone surveys of young women should include mobile dialling.</p