DNA methylation plays fundamental roles in gene regulation and cell identity in plants and animals. However, these two kingdoms exhibit distinct methylation patterns and reprogramming of methylation during development occurs in different ways. I have exciting evidence from the emerging model plant system Marchantia polymorpha of de novo DNA methylation at thousands of genes in the embryo that is analogous to gene methylation established in the oocyte of mammals. A subset of these genes are also methylated in the Marchantia male sex organ prior to fertilisation and are enriched for roles in hormone signalling, epigenetics, and developmental transitions, indicating parental imprinting events important for embryo development. I propose to conduct a targeted dissection of this phenomenon. I will be hosted first by Prof. Ecker (Salk Institute, US), who is a world leader in single cell epigenomics. I will carry out high-throughput single cell methylomics and transcriptomics in combination with other cutting-edge molecular techniques to precisely determine the developmental processes undergoing gene methylation reprogramming as well as the underlying epigenetic players involved. In the return phase, I will be hosted by Prof. Solano (Centro Nacional de Biotecnología, Spain) who is an expert in motif analysis and Marchantia hormone biology. With his expertise I will investigate the methylation targeting mechanism and will utilise methylation loss lines to confirm the role of gene methylation in transcriptional regulation as well as development, with a focus on ethylene signalling. This proposal represents a state-of-the-art class of study of DNA methylation through which I will broaden my scientific knowledge, develop complementary skills such as lab management and science outreach, and extend my professional network internationally. The experiences gained during this Fellowship will be essential to achieve my goal of leading a research group at the forefront of epigenetics.

Understanding how organisms adapt to their environments is a long-standing problem in Biology with far-reaching implications: adaptation affects the ability of species to survive in changing environments, host-pathogen interactions, and resistance to pesticides and drugs. Despite recent progress, adaptation is to date a poorly understood process largely due to limitations of current approaches that focus (i) on a priori candidate genes, (ii) on signals of selection at the DNA level without functional validation of the identified candidates, and (iii) on small sets of adaptive mutations that do not represent the variability present in natural populations. As a result, major questions such as what is the relative importance of different types of mutations in adaptation?, and what is the importance of epigenetic changes in adaptive evolution?, remain largely unanswered. To gain a deep understanding of adaptation, we need to systematically identify adaptive mutations across space and time, pinpoint their molecular mechanisms and discover their fitness effects. To this end, Drosophila melanogaster has proven to be an ideal organism. Besides the battery of genetic tools and resources available, D. melanogaster has recently adapted to live in out of Africa environments. We and others have already shown that transposable elements (TEs) have substantially contributed to the adaptation of D. melanogaster to different environmental challenges. Here, we propose to use state-of-the-art techniques, such as Illumina TruSeq sequencing and CRISPR/Cas9 genome editing, to systematically identify and characterize in detail adaptive TE insertions in D. melanogaster natural populations. Only by moving from gathering anecdotic evidence to applying global approaches, we will be able to start constructing a quantitative and predictive theory of adaptation that will be relevant for other species as well.

Neuronal loss is at the core of cognitive and functional failures of both acute brain injuries and neurodegenerative diseases. Direct neuronal reprogramming of local glial cells is emerging as a promising approach for restorative brain therapy. However, in order to use direct glia-to-neuron reprogramming for the treatment of neuronal loss we still need to address a number of challenges, namely reliable and long-term conversion into the desired neuron subtype. In this proposal we aim to generate specific neuronal subtypes using novel fate determinants in glia-to-neuron reprogramming, and to provide a detailed molecular analysis of the newly generated neurons over time. Our data indicate that ONECUT factors may represent excellent novel candidates for astrocyte reprogramming into neuronal fates. To address this possibility, in this proposal we will focus on the thalamocortical system, which represents the main input to the neocortex and it is essential to cortical processing. We hypothesize that the innovative combination of nuclei specific thalamic factors with ONECUT factors could reveal new avenues for the direct reprogramming of astrocytes into thalamic neurons of specific sensory modalities and may inform future strategies for brain repair. Here, we have unique expertise and molecular tools at hand that will allow us to reprogram astrocytes into specific neuron types in vitro and in vivo. Moreover, as our ultimate goal is to reprogram astrocytes to recover neuronal loss, we will test whether astrocytes from a sensory deprived thalamus can be reprogrammed. By using state-of-the-art techniques such as 3D light-sheet microscopy, calcium imaging and transcriptomic analysis, we will determine the fidelity and functionality of the newly generated neurons. This approach will offer us unparalleled advantages for the discovery of novel reprogramming combinations and address important questions about reliable and long-term conversion into the desired neuron type.

Catalysis is a key technology in the European economy, industry and sustainable growth for a resilient future. However, many catalytic methods in use today are not outstanding in fundamental aspects such as activity, selectivity, substrate scope, toxicity or even cost efficiency. The rich structural diversity and associated reactivity provided by heavy elements from the Main Group offer unique opportunities for the prospective substitution of traditional catalysts based on precious metals by less expensive, more abundant, and potentially less toxic Main Group compounds. Sub-valent Ge(II) and Sn(II) derivatives are particularly appealing due to their reduced HOMO-LUMO gaps and thus increased reactivity. Nevertheless, bond activation processes lead to very stable and unreactive products in higher oxidation states (+IV). As such, catalytic turnover via reductive regeneration of the active species is highly challenging. To address this problem, an interdisciplinary approach will be used combining fundamental aspects stemming from a priori independent areas of Chemistry. Rationally designed ambiphilic and bifunctional derivatives will be used as ligands for low-valent tetrylenes to induce cooperativity between Main Group elements and Transition Metals and promote novel reactivity in small molecule activation and functionalization. Another objective is to demonstrate that selective irradiation of well-designed mononuclear tetrylene will allow photoactivation of chemical bonds that are thermally inert towards many of these divalent species. However, we envision that the main advantage of using light will be in facilitating reductive elimination processes in the cases where thermal catalytic turnover is hampered by this rate-limiting step. For this reason, these studies will also be extended to our hybrid systems, in what constitutes a completely innovative research area, and where the possible synergies and cooperative mechanisms will be analyzed.