Novel Human Recombinant N-Acetylgalactosamine-6-Sulfate Sulfatase Produced in a Glyco-Engineered Escherichia coli Strain

Mucopolysaccharidosis IVA (MPS IVA) is a lysosomal storage disease caused by mutations in the gene encoding the lysosomal enzyme N-acetylgalactosamine-6-sulfate sulfatase (GALNS), resulting in the accumulation of keratan sulfate (KS) and chondroitin-6-sulfate (C6S). Previously, it was reported the production of an active human recombinant GALNS (rGALNS) in E. coli BL21(DE3). However, this recombinant enzyme was not taken up by HEK293 cells or MPS IVA skin fibroblasts. Here, we leveraged a glyco-engineered E. coli strain to produce a recombinant human GALNS bearing the eukaryotic trimannosyl core N-glycan, Man3GlcNAc2 (rGALNSoptGly). The N-glycosylated GALNS was produced at 100 mL and 1.65 L scales, purified and characterized with respect to pH stability, enzyme kinetic parameters, cell uptake, and KS clearance. The results showed that the addition of trimannosyl core N-glycans enhanced both protein stability and substrate affinity. rGALNSoptGly was capture through a mannose receptor-mediated process. This enzyme was delivered to the lysosome, where it reduced KS storage in human MPS IVA fibroblasts. This study demonstrates the potential of a glyco-engineered E. coli for producing a fully functional GALNS enzyme. It may offer an economic approach for the biosynthesis of a therapeutic glycoprotein that could prove useful for MPS IVA treatment. This strategy could be extended to other lysosomal enzymes that rely on the presence of mannose N-glycans for cell uptake

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality

Transcriptome profiling of liver of non-genetic low birth weight and long term health consequences

BACKGROUND: It is believed that the main factors of low prenatal growth in mammals are genetic and environmental. We used isogenic mice maintained in standard conditions to analyze how natural non-genetic microsomia (low birth weight) is produced in inbred mice and its long term effect on health. To better understand the molecular basis of non-genetic microsomia, we undertook transcriptome profiling of both male and female livers from small and normal size mice at birth. RESULTS: Naturally occurring neonatal microsomia was defined as a gender-specific weanling weight under the 10th percentile of the colony. Birth weight variation was similar in inbred and outbred lines. Mice were phenotyped by weight, size, blood pressure, organ size, their response to a glucose challenge, and survival rates. Regardless of diet, adult mice born with microsomia showed a significantly lower body weight and size, and differences in the weight of several organs of microsomic adult mice compared to normal birth weight adults were found. After a high-fat diet, microsomic mice were less prone to obesity, showing a better glucose tolerance and lower blood pressure. Through a transcriptome analysis, we detected a different pattern of mRNA transcription in the liver at birth comparing male vs female and microsomic vs normal mice, noting some modifications in epigenetic regulatory genes in females and modifications in some growth factor genes in males. Finally, using embryo transfer of embryos of different quality and age, we identified a putative preimplantation origin of this non-genetic microsomia. CONCLUSIONS: (1) neonatal microsomia is not always a risk factor for adult metabolic syndrome, (2) neonatal non-genetic microsomia displays changes in the expression of important epigenetic genes and changes in liver mRNA transcription profile at birth, exaggerating sexual dimorphism, and (3) random preimplantation phenotypic variability could partially explain body birth weight variation in isogenic lines

A high glucose concentration during early stages of in vitro equine embryo development alters expression of genes involved in glucose metabolism

9 Pág. Departamento de Reproducción animal (INIA)Equine embryos exhibit an unusual pattern of glucose tolerance in vitro and are currently cultured in hyperglycaemic conditions.This study was supported by the Spanish Ministry of Science, Innovation and Universities Competitiveness and Fondo Europeo de Desarrollo Regional (FEDER) (AEI/FEDER/UE); References: AGL2017‐84681‐R, RTI2018‐093548 and RYC‐2017‐21545. A.P.L.C acknowledges the financial support of the Universidad de Antioquia (COLCIENCIAS) through the Francisco José de Caldas Fellowship (512/2010).Peer reviewe

Anticoagulant selection in relation to the SAMe-TT2R2 score in patients with atrial fibrillation: The GLORIA-AF registry

Aim: The SAMe-TT2R2 score helps identify patients with atrial fibrillation (AF) likely to have poor anticoagulation control during anticoagulation with vitamin K antagonists (VKA) and those with scores >2 might be better managed with a target-specific oral anticoagulant (NOAC). We hypothesized that in clinical practice, VKAs may be prescribed less frequently to patients with AF and SAMe-TT2R2 scores >2 than to patients with lower scores. Methods and results: We analyzed the Phase III dataset of the Global Registry on Long-Term Oral Antithrombotic Treatment in Patients with Atrial Fibrillation (GLORIA-AF), a large, global, prospective global registry of patients with newly diagnosed AF and ≥1 stroke risk factor. We compared baseline clinical characteristics and antithrombotic prescriptions to determine the probability of the VKA prescription among anticoagulated patients with the baseline SAMe-TT2R2 score >2 and ≤ 2. Among 17,465 anticoagulated patients with AF, 4,828 (27.6%) patients were prescribed VKA and 12,637 (72.4%) patients an NOAC: 11,884 (68.0%) patients had SAMe-TT2R2 scores 0-2 and 5,581 (32.0%) patients had scores >2. The proportion of patients prescribed VKA was 28.0% among patients with SAMe-TT2R2 scores >2 and 27.5% in those with scores ≤2. Conclusions: The lack of a clear association between the SAMe-TT2R2 score and anticoagulant selection may be attributed to the relative efficacy and safety profiles between NOACs and VKAs as well as to the absence of trial evidence that an SAMe-TT2R2-guided strategy for the selection of the type of anticoagulation in NVAF patients has an impact on clinical outcomes of efficacy and safety. The latter hypothesis is currently being tested in a randomized controlled trial. Clinical trial registration: URL: https://www.clinicaltrials.gov//Unique identifier: NCT01937377, NCT01468701, and NCT01671007. © 2020 Hellenic Society of Cardiolog

Correction to: Comparative effectiveness and safety of non-vitamin K antagonists for atrial fibrillation in clinical practice: GLORIA-AF Registry

International audienceIn this article, the name of the GLORIA-AF investigator Anastasios Kollias was given incorrectly as Athanasios Kollias in the Acknowledgements. The original article has been corrected

Patterns of oral anticoagulant use and outcomes in Asian patients with atrial fibrillation: a post-hoc analysis from the GLORIA-AF Registry

Background: Previous studies suggested potential ethnic differences in the management and outcomes of atrial fibrillation (AF). We aim to analyse oral anticoagulant (OAC) prescription, discontinuation, and risk of adverse outcomes in Asian patients with AF, using data from a global prospective cohort study. Methods: From the GLORIA-AF Registry Phase II-III (November 2011-December 2014 for Phase II, and January 2014-December 2016 for Phase III), we analysed patients according to their self-reported ethnicity (Asian vs. non-Asian), as well as according to Asian subgroups (Chinese, Japanese, Korean and other Asian). Logistic regression was used to analyse OAC prescription, while the risk of OAC discontinuation and adverse outcomes were analysed through Cox-regression model. Our primary outcome was the composite of all-cause death and major adverse cardiovascular events (MACE). The original studies were registered with ClinicalTrials.gov, NCT01468701, NCT01671007, and NCT01937377. Findings: 34,421 patients were included (70.0 ± 10.5 years, 45.1% females, 6900 (20.0%) Asian: 3829 (55.5%) Chinese, 814 (11.8%) Japanese, 1964 (28.5%) Korean and 293 (4.2%) other Asian). Most of the Asian patients were recruited in Asia (n = 6701, 97.1%), while non-Asian patients were mainly recruited in Europe (n = 15,449, 56.1%) and North America (n = 8378, 30.4%). Compared to non-Asian individuals, prescription of OAC and non-vitamin K antagonist oral anticoagulant (NOAC) was lower in Asian patients (Odds Ratio [OR] and 95% Confidence Intervals (CI): 0.23 [0.22-0.25] and 0.66 [0.61-0.71], respectively), but higher in the Japanese subgroup. Asian ethnicity was also associated with higher risk of OAC discontinuation (Hazard Ratio [HR] and [95% CI]: 1.79 [1.67-1.92]), and lower risk of the primary composite outcome (HR [95% CI]: 0.86 [0.76-0.96]). Among the exploratory secondary outcomes, Asian ethnicity was associated with higher risks of thromboembolism and intracranial haemorrhage, and lower risk of major bleeding. Interpretation: Our results showed that Asian patients with AF showed suboptimal thromboembolic risk management and a specific risk profile of adverse outcomes; these differences may also reflect differences in country-specific factors. Ensuring integrated and appropriate treatment of these patients is crucial to improve their prognosis. Funding: The GLORIA-AF Registry was funded by Boehringer Ingelheim GmbH

Search for the lepton-flavour violating decays B0→K∗0μ±e∓B^0 \to K^{*0} \mu^\pm e^\mp and Bs0→ϕμ±e∓B_s^0 \to \phi \mu^\pm e^\mp

A search for the lepton-flavour violating decays $B^0 \to K^{*0} \mu^\pm e^\mp$ and $B_s^0 \to \phi \mu^\pm e^\mp$ is presented, using proton-proton collision data collected by the LHCb detector at the LHC, corresponding to an integrated luminosity of $9\,\text{fb}^{-1}$. No significant signals are observed and upper limits of \begin{align} {\cal B}( B^0 \to K^{*0} \mu^+ e^- ) &< \phantom{1}5.7\times 10^{-9}~(6.9\times 10^{-9}),\newline {\cal B}( B^0 \to K^{*0} \mu^- e^+ ) &< \phantom{1}6.8\times 10^{-9}~(7.9\times 10^{-9}),\newline {\cal B}( B^0 \to K^{*0} \mu^\pm e^\mp ) &< 10.1\times 10^{-9}~(11.7\times 10^{-9}),\newline {\cal B}( B_s^0 \to \phi \mu^\pm e^\mp ) &< 16.0\times 10^{-9}~(19.8\times 10^{-9}) \end{align} are set at $90\%~(95\%)$ confidence level. These results constitute the world's most stringent limits to date, with the limit on the decay $B_s^0 \to \phi \mu^\pm e^\mp$ the first being set. In addition, limits are reported for scalar and left-handed lepton-flavour violating New Physics scenarios.A search for the lepton-flavour violating decays B$^{0}$ → K$^{*0}$μ$^{±}$e$^{∓}$ and ${B}_s^0$→ ϕμ$^{±}$e$^{∓}$ is presented, using proton-proton collision data collected by the LHCb detector at the LHC, corresponding to an integrated luminosity of 9 fb$^{−1}$. No significant signals are observed and upper limits of${\displaystyle \begin{array}{c}\mathcal{B}\left({B}^0\to {K}^{\ast 0}{\mu}^{+}{e}^{-}\right)<5.7\times {10}^{-9}\left(6.9\times {10}^{-9}\right),\\ {}\mathcal{B}\left({B}^0\to {K}^{\ast 0}{\mu}^{-}{e}^{+}\right)<6.8\times {10}^{-9}\left(7.9\times {10}^{-9}\right),\\ {}\mathcal{B}\left({B}^0\to {K}^{\ast 0}{\mu}^{\pm }{e}^{\mp}\right)<10.1\times {10}^{-9}\left(11.7\times {10}^{-9}\right),\\ {}\mathcal{B}\left({B}_s^0\to \phi {\mu}^{\pm }{e}^{\mp}\right)<16.0\times {10}^{-9}\left(19.8\times {10}^{-9}\right)\end{array}}$are set at 90% (95%) confidence level. These results constitute the world’s most stringent limits to date, with the limit on the decay ${B}_s^0$→ ϕμ$^{±}$e$^{∓}$ the first being set. In addition, limits are reported for scalar and left-handed lepton-flavour violating New Physics scenarios.[graphic not available: see fulltext]A search for the lepton-flavour violating decays $B^0 \to K^{*0} \mu^\pm e^\mp$ and $B_s^0 \to \phi \mu^\pm e^\mp$ is presented, using proton-proton collision data collected by the LHCb detector at the LHC, corresponding to an integrated luminosity of $9\,\text{fb}^{-1}$. No significant signals are observed and upper limits of \begin{align} {\cal B}( B^0 \to K^{*0} \mu^+ e^- ) &< \phantom{1}5.7\times 10^{-9}~(6.9\times 10^{-9}),\newline {\cal B}( B^0 \to K^{*0} \mu^- e^+ ) &< \phantom{1}6.8\times 10^{-9}~(7.9\times 10^{-9}),\newline {\cal B}( B^0 \to K^{*0} \mu^\pm e^\mp ) &< 10.1\times 10^{-9}~(11.7\times 10^{-9}),\newline {\cal B}( B_s^0 \to \phi \mu^\pm e^\mp ) &< 16.0\times 10^{-9}~(19.8\times 10^{-9}) \end{align} are set at $90\%~(95\%)$ confidence level. These results constitute the world's most stringent limits to date, with the limit on the decay $B_s^0 \to \phi \mu^\pm e^\mp$ the first being set. In addition, limits are reported for scalar and left-handed lepton-flavour violating New Physics scenarios