Biology subjects

Esteban, A.

Publications and source records attributed to Esteban, A..

4 recordsLinked to original sources

bioRxiv2021

Alu-mediated weak CEBPA binding and slow B cell transdifferentiation in human

Many developmental and differentiation processes take substantially longer in human than in mouse. To investigate the molecular mechanisms underlying this phenomenon, here we have specifically focused on the transdifferentiation from B cells to macrophages. The process is triggered by exactly the same molecular mechanism -- the induction by the transcription factor (TF) CEBPA -- but takes three days in mouse and seven in human (1, 2). In mouse, the speed of this process is known to be associated with Myc expression (3). We found that in this species, CEBPA binds strongly to the Myc promoter, efficiently down-regulating Myc. In human, in contrast, CEBPA does not bind this promoter, and MYC is indirectly and more slowly down-regulated. Attenuation of CEBPA binding is not specific to the MYC promoter, but a general trait of the human genome across multiple biological conditions. We traced back weak CEBPA binding to the primate-specific Alu repeat expansion. Many Alu repeats carry strong CEBPA binding motifs, which sequester CEBPA, and attenuate CEBPA binding genome-wide. We observed similar CEBPA and MYC dynamics in natural processes regulated by CEBPA, suggesting that CEBPA attenuation could underlie the longer duration in human processes controlled by this factor. Our work highlights the highly complex mode in which biological information is encoded in genome sequences, evolutionarily connecting, in an unexpected way, lineage-specific transposable element expansions to species-specific changes in developmental tempos.

bioinformatics↗

bioRxiv2021

Paired guide RNA CRISPR-Cas9 screening for protein-coding genes and lncRNAs involved in transdifferentiation of human B-cells to macrophages

CRISPR-Cas9 screening libraries have arisen as a powerful tool to identify both protein coding (pc) and non-coding genes playing a role along different processes. In particular, the usage of a nuclease active Cas9 coupled to a single gRNA has proven to efficiently impair the expression of pc-genes by generating deleterious frameshifts. Here, we first demonstrate that the usage of a second gRNA targeting the same gene synergistically enhances the capacity of the CRISPR-Cas9 system to knock out pc-genes. We next take advantage of our paired-guide (pgRNA) system to design a library to simultaneously target 874 pc-genes and 166 lncRNAs which are known to change expression during the transdifferentiation from pre-B cells to macrophages. We show that this system is able to identify known players in this process, and also predicts 26 potential novel ones, of which we select four for deeper characterization. Two of these, FURIN and NFE2, code for proteins related to cell differentiation and macrophage function; the other two, LINC02432 and MIR3945HG, are lncRNAs associated with cancerous and infectious diseases, respectively. The CRISPR-Cas9 coupled to pgRNAs system is, therefore, a suitable tool to target simultaneously pc-genes and lncRNAs for genomic perturbation assays.

genomics↗

bioRxiv2020

Dynamics of gene expression and chromatin marking during cell state transition

We have monitored the transcriptomic and epigenomic status of cells at twelve time-points during the transdifferentiation of human pre-B cells into macrophages. Using this data, we have investigated some fundamental questions regarding the role of chromatin in gene expression. We have found that, over time, genes are characterized by a limited number of chromatin states (combinations of histone modifications), and that, consistently, chromatin changes over genes tend to occur in a coordinated manner. We have observed strong association between these changes and gene expression only at the time of initial gene activation. Activation is preceded by H3K4me1 and H3K4me2, and followed in a precise order by most other histone modifications. Further changes in gene expression, comparable or even stronger than those at initial activation, occur without associated changes in histone modifications. The data generated here constitutes, thus, a unique resource to investigate transcriptomic and epigenomic dynamics during a differentiation process.

genomics↗

bioRxiv2019

Catching the big picture of the Mediterranean Sea biodiversity with an end-to-end model of climate and fishing impacts

The Mediterranean Sea is among the main hotspots of marine biodiversity in the world. Under combined pressures of fishing activities and climate change it has also become a hotspot of global change, with increased concern about the worsening status of marine exploited species. More integrated modelling approaches are needed to anticipate global change impacts in the Mediterranean Sea, in order to help decision makers prioritizing management actions and strategies, mitigating impacts and adapting to changes. Our challenge was to develop a holistic model of the marine biodiversity in the Mediterranean Sea with an explicit representation of the spatial multispecies dynamics of exploited resources under the combined influence of climate variability and fishing pressure. An individual-based ecosystem model OSMOSE (Object-oriented Simulator of Marine ecOSystEms), including 100 marine species (fish, cephalopods and crustaceans) and representing about 95 % of the total declared catches, has been implemented for the first time at a high spatial resolution (400 km2) and at a large spatial scale (whole Mediterranean basin). The coupling of OSMOSE to the NEMOMED 12 physical model, and to the Eco3M-S biogeochemical and low trophic level model has been achieved to build the OSMOSE-MED end-to-end model. We fitted OSMOSE-MED to observed and estimated data of biomass and commercial catches using a likelihood approach and an evolutionary optimization algorithm. The outputs of OSMOSE-MED were then verified against observed biomass and catches, and confronted to independent datasets (MEDITS data, diet compositions and trophic levels). Although some improvements are suggested for future developments, the model results at different hierarchical levels, from individuals up to the ecosystem scale, were consistent with current knowledge and observations on the structure, the functioning and the dynamics of the ecosystems in the Mediterranean Sea. All the modelling steps, from the comprehensive representation of key ecological processes and feedbacks, the careful parameterization of the model, the confrontation to observed data, and the positive outcome from the validation process, allowed to strengthen the degree of realism of OSMOSE-MED and its relevance as an impact model to explore the futures of marine biodiversity under scenarios of global change, and as a tool to support the implementation of ecosystem-based fisheries management in the Mediterranean Sea.

ecology↗