Mutant CEBPA directly drives the expression of the targetable tumor-promoting factor CD73 in AML

The key myeloid transcription factor (TF), CEBPA, is frequently mutated in acute myeloid leukemia (AML), but the direct molecular effects of this leukemic driver mutation remain elusive. To investigate mutant AML, we performed microscale, in vivo chromatin immunoprecipitation sequencing and identified a set of aberrantly activated enhancers, exclusively occupied by the leukemia-associated CEBPA-p30 isoform. Comparing gene expression changes in human mutant AML and the corresponding mouse model, we identified , encoding CD73, as a cross-species AML gene with an upstream leukemic enhancer physically and functionally linked to the gene. Increased expression of CD73, mediated by the CEBPA-p30 isoform, sustained leukemic growth via the CD73/A2AR axis. Notably, targeting of this pathway enhanced survival of AML-transplanted mice. Our data thus indicate a first-in-class link between a cancer driver mutation in a TF and a druggable, direct transcriptional target

A new genetic tool to improve immune-compromised mouse models : Derivation and CRISPR/Cas9-mediated targeting of NRG embryonic stem cell lines

Development of human hematopoietic stem cells and differentiation of embryonic stem (ES) cells/induced pluripotent stem (iPS) cells to hematopoietic stem cells are poorly understood. NOD (Non-obese diabetic)-derived mouse strains, such as NSG (NOD-Scid-il2Rg) or NRG (NOD-Rag1-il2Rg), are the best available models for studying the function of fetal and adult human hematopoietic cells as well as ES/iPS cell-derived hematopoietic stem cells. Unfortunately, engraftment of human hematopoietic stem cells is very variable in these models. Introduction of additional permissive mutations into these complex genetic backgrounds of the NRG/NSG mice by natural breeding is a very demanding task in terms of time and resources. Specifically, since the genetic elements defining the NSG/NRG phenotypes have not yet been fully characterized, intense backcrossing is required to ensure transmission of the full phenotype. Here we describe the derivation of embryonic stem cell (ESC) lines from NRG pre-implantation embryos generated by in vitro fertilization followed by the CRISPR/CAS9 targeting of the Gata-2 locus. After injection into morula stage embryos, cells from three tested lines gave rise to chimeric adult mice showing high contribution of the ESCs (70%–100%), assessed by coat color. Moreover, these lines have been successfully targeted using Cas9/CRISPR technology, and the mutant cells have been shown to remain germ line competent. Therefore, these new NRG ESC lines combined with genome editing nucleases bring a powerful genetic tool that facilitates the generation of new NOD-based mouse models with the aim to improve the existing xenograft models

ELIXIR-CONVERGE D5.4 Report on KPI

The overall goal of ELIXIR-CONVERGE&rsquo;s WP5 was, based on the analysis of a set of very diverse use cases, to contribute to the development of a set of resources supporting the development and implementation of Data Management Plans (DMPs) in national and transnational projects. Six use cases were considered to co-develop and test a method to address domain specific data management planning associated to a set of resources in collaboration with WP1, WP2 and WP3. In this context, another objective of WP5 was to develop, implement and refine key performance indicators (KPIs) in order to monitor the demonstrator projects&rsquo; implementation of data management plans and possibly assess their adoption by the relevant community. WP5 developed two sets KPIs in collaboration with WP4, in charge of developing KPIs across the entire ELIXIR-CONVERGE project and for assessing the impact of the project : a first set aiming at monitoring the development of guidance, resources for the development of DMP and for training in the context of the use cases a second set aiming at addressing the adoption of these resources by relevant communities of users as a way to assess the impact of the work achieved In parallel, in order to ensure long term maintenance/update of the developed resources and to increase their impact, WP5 started to engage with ELIXIR communities that could be natural owners of these resources. The KPI developed and collected by WP5 during the ELIXIR-CONVERGE project were useful to follow the partner&rsquo;s progress in the development and test of a sort of starter kit for domain specific data management support. Success stories could be collected showing adoption by communities and new projects and were mapped on ELIXIR&rsquo;s categories of impacts showing already &ldquo;hits&rdquo; on several of these.</p

The European Genome-phenome Archive in 2021

The European Genome-phenome Archive (EGA - https://ega-archive.org/) is a resource for long term secure archiving of all types of potentially identifiable genetic, phenotypic, and clinical data resulting from biomedical research projects. Its mission is to foster hosted data reuse, enable reproducibility, and accelerate biomedical and translational research in line with the FAIR principles. Launched in 2008, the EGA has grown quickly, currently archiving over 4,500 studies from nearly one thousand institutions. The EGA operates a distributed data access model in which requests are made to the data controller, not to the EGA, therefore, the submitter keeps control on who has access to the data and under which conditions. Given the size and value of data hosted, the EGA is constantly improving its value chain, that is, how the EGA can contribute to enhancing the value of human health data by facilitating its submission, discovery, access, and distribution, as well as leading the design and implementation of standards and methods necessary to deliver the value chain. The EGA has become a key GA4GH Driver Project, leading multiple development efforts and implementing new standards and tools, and has been appointed as an ELIXIR Core Data Resource.Horizon 2020 Programme of the European Union [CORBEL [654248], ELIXIR-EXCELERATE [676559], Solve-RD [779257], EASI-Genomics [824110], EJP-RD [825575], CINECA [825775], EuCanCan [825835], EUCanshare [825903], ELIXIR-CONVERGE [871075]]; Wellcome Trust Global Alliance for Genomics and Health [201535/Z/16/Z]; UK Biobank; Chan Zuckerberg Initiative DAF, an advised fund of Silicon Valley Community Foundation [2017-171304 (5022)]; European Molecular Biology Laboratory (EMBL); LaCaixa Foundation [004745/008034]; [LCF/PR/CE20/50740008]. Funding for open access charge: LaCaixa Foundation [LCF/PR/CE20/50740008]

Hematopoietic stem cell development requires transient Wnt/β-catenin activity

Understanding how hematopoietic stem cells (HSCs) are generated and the signals that control this process is a crucial issue for regenerative medicine applications that require in vitro production of HSC. HSCs emerge during embryonic life from an endothelial-like cell population that resides in the aorta-gonad-mesonephros (AGM) region. We show here that β-catenin is nuclear and active in few endothelial nonhematopoietic cells closely associated with the emerging hematopoietic clusters of the embryonic aorta during mouse development. Importantly, Wnt/β-catenin activity is transiently required in the AGM to generate long-term HSCs and to produce hematopoietic cells in vitro from AGM endothelial precursors. Genetic deletion of β-catenin from the embryonic endothelium stage (using VE-cadherin–Cre recombinase), but not from embryonic hematopoietic cells (using Vav1-Cre), precludes progression of mutant cells toward the hematopoietic lineage; however, these mutant cells still contribute to the adult endothelium. Together, those findings indicate that Wnt/β-catenin activity is needed for the emergence but not the maintenance of HSCs in mouse embryos

A new accelerated laboratory test for the assessment of the durability of materials with respect to salt crystallization

The RILEM Technical Committee 271-ASC was set up in 2016 with the aim of developing an improved procedure for the assessment of the durability of porous building materials, such as brick and natural stone, against salt crystallization, accelerating the deterioration process without significantly altering its mechanism. The test procedure developed by the TC 271-ASC proposes a new approach to salt crystallization tests. It starts from the consideration that it is necessary to accumulate a certain amount of salt to activate the damage. Thus salt damage can be seen as a process developing in two phases: accumulation and propagation. Based on this approach, a new salt crystallization test procedure has been defined, consisting of two phases: a first phase, in which salts are introduced in the material and accumulate close to the evaporation surface, followed by a second phase, in which damage propagates because of repeated dissolution and crystallization cycles induced by re-wetting with liquid water and by relative humidity (RH) changes. In this paper the procedure is described and the reasons for the choices made are elucidated. The procedure has been tested on two types of limestone and, at the moment of writing, is being validated in a round robin test carried out on 9 different substrates and involving 11 laboratories. Based on the results of the round robin test, the procedure will be fine-tuned