Epitope-engineered human hematopoietic stem cells are shielded from CD123-targeted immunotherapy

Targeted eradication of transformed or otherwise dysregulated cells using monoclonal antibodies (mAb), antibody-drug conjugates (ADC), T cell engagers (TCE), or chimeric antigen receptor (CAR) cells is very effective for hematologic diseases. Unlike the breakthrough progress achieved for B cell malignancies, there is a pressing need to find suitable antigens for myeloid malignancies. CD123, the interleukin-3 (IL-3) receptor alpha-chain, is highly expressed in various hematological malignancies, including acute myeloid leukemia (AML). However, shared CD123 expression on healthy hematopoietic stem and progenitor cells (HSPCs) bears the risk for myelotoxicity. We demonstrate that epitope-engineered HSPCs were shielded from CD123-targeted immunotherapy but remained functional, while CD123-deficient HSPCs displayed a competitive disadvantage. Transplantation of genome-edited HSPCs could enable tumor-selective targeted immunotherapy while rebuilding a fully functional hematopoietic system. We envision that this approach is broadly applicable to other targets and cells, could render hitherto undruggable targets accessible to immunotherapy, and will allow continued posttransplant therapy, for instance, to treat minimal residual disease (MRD)

The science behind soft skills: Do’s and Don’ts for early career researchers and beyond. A review paper from the EU-CardioRNA COST Action CA17129

peer reviewedSoft skills are the elementary management, personal, and interpersonal abilities that are vital for an individual to be efficient at workplace or in their personal life. Each work place requires different set of soft skills. Thus, in addition to scientific/technical skills that are easier to access within a short time frame, several key soft skills are essential for the success of a researcher in today’s international work environment. In this paper, the trainees and trainers of the EU-CardioRNA COST Action CA17129 training school on soft skills present basic and advanced soft skills for early career researchers. Here, we particularly emphasize on the importance of transferable and presentation skills, ethics, literature reading and reviewing, research protocol and grant writing, networking, and career opportunities for researchers. All these skills are vital but are often overlooked by some scholars. We also provide tips to ace in aforementioned skills that are crucial in a day-to-day life of early and late career researchers in academia and industry.</ns4:p

Relationship between circulating microRNA-30c with total- and LDL-cholesterol, their circulatory transportation and effect of statins

Background: Small non-coding microRNAs (miR) have important regulatory roles and are used as biomarkers of disease. We investigated the relationship between lipoproteins and circulating miR-30c, evaluated how they are transported in circulation and determined whether statins altered the circulating concentration of miR-30c. Methods: To determine the relationship between lipoproteins and circulating miR-30c, serum samples from 79 subjects recruited from a lipid clinic were evaluated. Ultracentrifugation and nanoparticle tracking analysis was used to evaluate the transportation of miR-30c in the circulation by lipoproteins and extracellular vesicles in three healthy volunteers. Using archived samples from previous studies, the effects of 40 mg rosuvastatin (n = 22) and 40 mg pravastatin (n = 24) on miR-30c expression was also examined. RNA extraction, reverse transcription-quantitative real-time polymerase chain reaction was carried out using standard procedures. Results: When stratified according to total cholesterol concentration, there was increased miR-30c expression in the highest compared to the lowest tertile (p = 0.035). There was significant positive correlation between miR- 30c and total- (r = 0.367; p = 0.002) and LDL-cholesterol (r = 0.391; p = 0.001). We found that miR-30c was transported in both exosomes and on HDL3. There was a 3.8-fold increased expression of circulating miR-30c after pravastatin treatment for 1 year (p = 0.005) but no significant change with atorvastatin after 8 weeks (p = 0.145). Conclusions: This study shows for the first-time in humans that circulating miR-30c is significantly, positively correlated with total- and LDL-cholesterol implicating regulatory functions in lipid homeostasis. We show miR-30c is transported in both exosomes and on HDL3 and pravastatin therapy significantly increased circulating miR-30c expression adding to the pleiotropic dimensions of statins

Anti-NF155 chronic inflammatory demyelinating polyradiculoneuropathy strongly associates to HLA-DRB15

Background: The aim of the research is to study the human leukocyte antigen (HLA) class II allele frequencies in chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) associated with anti-neurofascin 155 (NF155) antibodies. Methods: Thirteen anti-NF155+ and 35 anti-NF155 negative (anti-NF155neg) CIDP patients were included in a casecontrol study. The frequencies of the DRB1 HLA allele were analyzed in all patients while DQ frequencies were only studied in patients sharing the DRB1*15 allele. In silico HLA-peptide binding and NF155 antigenicity, predictions were performed to analyze overlap between presented peptides and antigenic regions. Results: DRB1*15 alleles (DRB1*15:01 and DRB1*15:02) were present in 10 out of 13 anti-NF155+ CIDP patients and in only 5 out of 35 anti-NF155neg CIDP patients (77 vs 14%; OR = 20, CI = 4.035 to 99.13). DRB1*15 alleles appeared also in significantly higher proportions in anti-NF155+ CIDP than in normal population (77 vs 17%; OR = 16.9, CI = 4.434 to 57. 30). Seven anti-NF155+ CIDP patients (53%) and 5 anti-NF155neg CIDP patients had the DRB1*15:01 allele (OR = 7, p = 0.009), while 3 anti-NF155+ CIDP patients and none of the anti-NF155neg CIDP patients had the DRB1*15:02 allele (OR = 23.6, p = 0.016). In silico analysis of the NF155 peptides binding to DRB1*15 alleles showed significant overlap in the peptides presented by the 15:01 and 15:02 alleles, suggesting functional homology. Conclusions: DRB1*15 alleles are the first strong risk factor associated to a CIDP subset, providing additional evidence that anti-NF155+ CIDP patients constitute a differentiated disease within the CIDP syndrome

Global prevalence and genotype distribution of hepatitis C virus infection in 2015 : A modelling study

Publisher Copyright: © 2017 Elsevier LtdBackground The 69th World Health Assembly approved the Global Health Sector Strategy to eliminate hepatitis C virus (HCV) infection by 2030, which can become a reality with the recent launch of direct acting antiviral therapies. Reliable disease burden estimates are required for national strategies. This analysis estimates the global prevalence of viraemic HCV at the end of 2015, an update of—and expansion on—the 2014 analysis, which reported 80 million (95% CI 64–103) viraemic infections in 2013. Methods We developed country-level disease burden models following a systematic review of HCV prevalence (number of studies, n=6754) and genotype (n=11 342) studies published after 2013. A Delphi process was used to gain country expert consensus and validate inputs. Published estimates alone were used for countries where expert panel meetings could not be scheduled. Global prevalence was estimated using regional averages for countries without data. Findings Models were built for 100 countries, 59 of which were approved by country experts, with the remaining 41 estimated using published data alone. The remaining countries had insufficient data to create a model. The global prevalence of viraemic HCV is estimated to be 1·0% (95% uncertainty interval 0·8–1·1) in 2015, corresponding to 71·1 million (62·5–79·4) viraemic infections. Genotypes 1 and 3 were the most common cause of infections (44% and 25%, respectively). Interpretation The global estimate of viraemic infections is lower than previous estimates, largely due to more recent (lower) prevalence estimates in Africa. Additionally, increased mortality due to liver-related causes and an ageing population may have contributed to a reduction in infections. Funding John C Martin Foundation.publishersversionPeer reviewe

Mitochondrial physiology

As the knowledge base and importance of mitochondrial physiology to evolution, health and disease expands, the necessity for harmonizing the terminology concerning mitochondrial respiratory states and rates has become increasingly apparent. The chemiosmotic theory establishes the mechanism of energy transformation and coupling in oxidative phosphorylation. The unifying concept of the protonmotive force provides the framework for developing a consistent theoretical foundation of mitochondrial physiology and bioenergetics. We follow the latest SI guidelines and those of the International Union of Pure and Applied Chemistry (IUPAC) on terminology in physical chemistry, extended by considerations of open systems and thermodynamics of irreversible processes. The concept-driven constructive terminology incorporates the meaning of each quantity and aligns concepts and symbols with the nomenclature of classical bioenergetics. We endeavour to provide a balanced view of mitochondrial respiratory control and a critical discussion on reporting data of mitochondrial respiration in terms of metabolic flows and fluxes. Uniform standards for evaluation of respiratory states and rates will ultimately contribute to reproducibility between laboratories and thus support the development of data repositories of mitochondrial respiratory function in species, tissues, and cells. Clarity of concept and consistency of nomenclature facilitate effective transdisciplinary communication, education, and ultimately further discovery

Mitochondrial physiology

As the knowledge base and importance of mitochondrial physiology to evolution, health and disease expands, the necessity for harmonizing the terminology concerning mitochondrial respiratory states and rates has become increasingly apparent. The chemiosmotic theory establishes the mechanism of energy transformation and coupling in oxidative phosphorylation. The unifying concept of the protonmotive force provides the framework for developing a consistent theoretical foundation of mitochondrial physiology and bioenergetics. We follow the latest SI guidelines and those of the International Union of Pure and Applied Chemistry (IUPAC) on terminology in physical chemistry, extended by considerations of open systems and thermodynamics of irreversible processes. The concept-driven constructive terminology incorporates the meaning of each quantity and aligns concepts and symbols with the nomenclature of classical bioenergetics. We endeavour to provide a balanced view of mitochondrial respiratory control and a critical discussion on reporting data of mitochondrial respiration in terms of metabolic flows and fluxes. Uniform standards for evaluation of respiratory states and rates will ultimately contribute to reproducibility between laboratories and thus support the development of data repositories of mitochondrial respiratory function in species, tissues, and cells. Clarity of concept and consistency of nomenclature facilitate effective transdisciplinary communication, education, and ultimately further discovery

Innovation et développement dans les systèmes agricoles et alimentaires

L’innovation est souvent présentée comme l’un des principaux leviers pour promouvoir un développement plus durable et plus inclusif. Dans les domaines de l’agriculture et de l’alimentation, l’innovation est marquée par des spécificités liées à sa relation à la nature, mais aussi à la grande diversité d’acteurs concernés, depuis les agriculteurs jusqu’aux consommateurs, en passant par les services de recherche et de développement. L’innovation émerge des interactions entre ces acteurs, qui mobilisent des ressources et produisent des connaissances dans des dispositifs collaboratifs, afin de générer des changements. Elle recouvre des domaines aussi variés que les pratiques de production, l’organisation des marchés, ou les pratiques alimentaires. L’innovation est reliée aux grands enjeux de développement : innovation agro-écologique, innovation sociale, innovation territoriale, etc. Cet ouvrage porte un regard sur l’innovation dans les systèmes agricoles et alimentaires. Il met un accent particulier sur l’accompagnement de l’innovation, en interrogeant les méthodes et les organisations, et sur l’évaluation de l’innovation au regard de différents critères. Il s’appuie sur des réflexions portées par différentes disciplines scientifiques, sur des travaux de terrain conduits tant en France que dans de nombreux pays du Sud, et enfin sur les expériences acquises en accompagnant des acteurs qui innovent. Il combine des synthèses sur l’innovation et des études de cas emblématiques pour illustrer les propos. L’ouvrage est destiné aux enseignants, professionnels, étudiants et chercheurs

Role of c-di-AMP in the physiology of Streptococcus agalactiae

Streptococcus agalactiae, également appelé streptocoque du groupe B (SGB) est une bactérie à Gram positif, capsulée, commensale des voies digestives et urogénitales. Cependant, le SGB est la principale cause d’infections invasives (septicémie, pneumonie et méningite) chez les nouveau-nés dans les pays occidentaux à haut revenu (UE, Amériques du nord). La virulence du SGB dépend de plusieurs facteurs impliqués dans l’adhésion et l’invasion tissulaire, ainsi que dans l’échappement au système immunitaire de l’hôte. Récemment, un nouveau mécanisme a été identifié au sein de l’Unité permettant au SGB de contrôler l’activation du système immunitaire inné par le di-AMP cyclique (di-AMPc) bactérien. Le di-AMPc est synthétisé spécifiquement par les bactéries et est reconnu par les cellules de l'hôte infectées pour induire une réponse immunitaire innée. Le SGB échappe à cette réponse immunitaire grâce à l'expression d'une enzyme ancrée à sa paroi, l’ecto-nucléotidase CdnP, qui est capable de dégrader le di-AMPc. La description du rôle du di-AMPc dans la relation hôte-pathogène nous a amené à étudier son rôle dans la physiologie du SGB. Au cours de ma thèse j'ai caractérisé le gène dacA codant pour la seule enzyme synthétisant le di-AMPc dans le SGB. L’inactivation conditionnelle de dacA a permis de démontrer que la synthèse de di-AMPc est essentielle à la croissance du SGB dans des conditions standards de culture, i.e. culture en milieu riche et en aérobie. En testant différentes conditions de croissance, j’ai observé que la synthèse de di-AMPc n’est plus essentielle lors d'une culture en milieu riche en anaérobie. Ainsi, la délétion du gène dacA a été obtenue dans ces conditions permissives. Les mutants dacA obtenus sont génétiquement très instables et accumulent des mutations secondaires compensatrices. Ces mutations ont été identifiées par séquençage et leurs effets confirmés par des tests fonctionnels de ré-expression de l'allèle original des gènes mutés. Globalement, les mutations permettant de compenser l’absence de synthèse de di-AMPc sont localisées dans des systèmes impliqués dans le maintien de la pression osmotique interne. Par une approche biochimique ciblée, les principales protéines fixant le di-AMPc ont été identifiées, parmi lesquelles des transporteurs de potassium, d’osmolytes, et un facteur de transcription original impliqué dans l’osmorégulation. A partir des approches génétique et biochimique, j’ai pu définir des conditions de culture dans lesquelles la synthèse de di-AMPc n’est plus nécessaire à la croissance i.e. culture dans un milieu synthétique en limitant l’apport de potassium et d’osmolytes. Ainsi, j’ai pu identifier la fonction essentielle du di-AMPc dans la régulation de la pression osmotique interne avec un rôle prépondérant des transporteurs d’osmolytes de type bétaïne. De façon intéressante, cette fonction est conservée dans les bactéries, mais les mécanismes mis en jeu diffèrent selon les espèces considérées, reflétant probablement une adaptation spécifique au métabolisme et aux conditions environnementales rencontrées par ces bactéries.Streptococcus agalactiae, also known as group B streptococcus (GBS) is a Gram-positive encapsulated bacterium, commensal of the intestinal and vaginal tracts. However, GBS is also the leading cause of neonatal invasive infections (septicaemiae, pneumonia and meningitis) in western countries (EU, North America). GBS virulence depend on several factors involved in adhesion and invasion of host cells, as well as evasion to host immunity. Recently, a novel mechanism was identified by the Unit allowing GBS to control the activation of innate immune system by bacterial cyclic di-AMP (c-di-AMP). c-di-AMP is specifically synthetized by bacteria and is recognized by host infected cells to induce an innate immune response. GBS escape to this immune response by expressing a cell wall anchored ectonucleotidase CdnP that degrade c-di-AMP. Given the role of c-di-AMP in host-pathogen interactions we characterized its role in bacterial physiology. During my PhD I have characterized dacA encoding for the unique enzyme that synthetized c-di-AMP in GBS. Conditional inactivation of dacA demonstrated that c-di-AMP is essential for GBS growth on rich medium in aerobiosis. By testing different growth conditions, I observed that c-di-AMP synthesis is dispensable on rich medium in anaerobiosis. This permissive condition was used to obtain dacA deletion mutants. The dacA mutants are genetically unstable and accumulate compensatory mutations. These mutations were identified by whole genome sequencing and their effects confirmed by re-expressing the WT allele of the mutated genes. Overall, the mutations allowing to compensate growth without c-di-AMP are localized in systems involved in the control of osmotic pressure. By a targeted biochemical approach, the main c-di-AMP binding proteins were identified, among them potassium and osmolyte transporters as well as an original transcription factor involved in osmoregulation. From the results of genetic and targeted biochemical approaches, I defined growth conditions where c-di-AMP synthesis is no more essential i.e. culture in synthetic medium by limiting potassium and osmolytes addition. Thus, I identified the essential function of c-di-AMP in the regulation of osmotic pressure with a major role of betain-type osmolytes transporters. Interestingly, this function is conserved in bacteria but different species-specific mechanisms are involved, this might reflect the metabolic adaptation of bacteria to their specific environments