Cells in our bodies constantly experience mechanical forces from their microenvironment. When cells sense a critical threshold of elevated tension, they hold tight together and allow tissues to function healthily as a group. In certain diseases, however, our cells lose their mechanosensing and adhesive properties and, as a result they get dissociated, as in the case of muscular dystrophies. Integrin-based adhesions to the extracellular matrix (ECM) are emerging as key networks of mechanotransmission. This proposal aims to discover how mechanical forces modulate cell-matrix adhesion at the myotendinous junctions in the developing Drosophila embryo, combining biophysical, molecular and genetic approaches. To achieve this goal, I propose to implement two complementary specific objectives: First, I will identify and quantify the relationship between forces and adhesion strength in mutants affecting either integrin-ECM binding or muscle contractility by utilizing in vivo laser ablation and magnetic tweezers. Second, I will examine whether and how IPP complex -a core module of the integrin adhesome- alters the molecular forces transmitted across Talin, which is a major mechanosensor at integrin junctions, utilizing suitable FRET-based biosensors. Collectively, this interdisciplinary research will provide a novel mechanical framework of how cells integrate forces and maintain tissue integrity in the living organism. Given the striking similarities in the molecular organisation of the myotendinous junctions between fly and human, the outcome of this work will provide a deeper understanding of how we can better combat dystrophic diseases.

It is estimated that over 36 million Europeans suffer from an autoimmune disease (AID) and according to the European League against Rheumatism one third of people of all ages are affected at some point during their lifetime. AIDs are a significant clinical problem, however, most of the current therapies target the terminal phase of inflammation and do not suppress the fundamental events responsible for the initiation of the autoimmune process. The EXAUTOIMMUNE project will use Sjögren’s syndrome (SS), one of the most common AIDs, as a model to create innovating strategies for dissecting the fundamental processes involved in the initiation of the autoimmune response. By using state-of-the-art nanotechnology we will characterize the extracellular vesicles (EV) secreted by the salivary gland epithelial cells, the main target of autoimmune responses in SS, and test their immunoregulatory properties. Then, we will apply next-generation sequencing to analyze the EVs’ RNA content and identify candidate RNA molecules that can be functionally tested for disruption of autoimmune processes. Linking EVs and extracellular RNA to autoimmunity is a fundamental theme of the current knowledge. The results of the proposed research will substantially add to the delineation of the biological relevance of extracellular RNA in modulating the regulatory mechanisms leading to autoimmunity. This study will form the foundation for transitioning from human biology to a mouse model, where the potential therapeutic value of EV and/or RNA-targeting will be tested, thus opening the best career opportunities for the applicant as a leader in this newly emerging landscape. Spearheading this research using next-generation tools and ideas will produce long-term synergies and will define EU and ERA as world leaders in the battle against AID. Collectively, the project activities will open the best career opportunities for the candidate, while advancing EU’s scientific competitiveness and innovation.

Cancer is one of the leading causes of death worldwide and its management remains challenging. Tumor evolution, the process of genetic and epigenetic divergence and selection of cancer cell clones in a heterogeneous tumor, is a key aspect of disease progression and therapy resistance. The main goal of this proposal is to expand our understanding of the biological mechanisms involved in these processes. So far, the role of chromatin regulation and epigenetics in tumor evolution remains poorly characterized and understood. This is particularly relevant in bladder cancer where mutations in chromatin regulation are very frequent and are associated with advanced and therapy resistant disease. Epi-BCE will advance the state-of-the-art through the discovery of epigenetic mechanisms that drive bladder cancer progression and chemoresistance in cell subpopulations, and tumors with chromatin deregulation. Specifically, I will utilize a carcinogen-based mouse model of bladder cancer that recapitulates human cancer stages from early to advanced and therapy resistant disease. I will perform transcriptomics and epigenomics at bulk and single-cell level at stages of disease progression. In addition, I will employ genetically engineered mice with ablation of chromatin regulation proteins, integrative computational analysis and functional genetic experiments to identify key mechanisms. Epi-BCE will be a platform for professional growth. I will receive technical training in experimental mouse models, bioinformatics, and 3D genomics through a secondment. I will engage in new professional networks and acquire transferable skills. Grantsmanship activities will support the fellow towards a path to independence. Overall, the proposed experiments will yield scientific knowledge and reusable datasets on cancer biology. These outcomes will promote and accelerate future research on cancer management and therapeutics.