Autoimmune encephalitides (AE) are severe brain disorders mediated by antibodies against neuronal proteins, such as the N-Methyl-D-Aspartate receptor (NMDAR). The antibody effects are reversible, explaining recovery after antibody-depleting therapies. However, it is unknown why many patients have incomplete recovery and persistent deficits. In these cases, additional immune mechanisms might be at play. The objective of ImmuBRAIN is to define how the adaptive immune response (autoantibodies) cross-talks with innate immune-mechanisms, resulting in neuronal dysfunction or death, by using NMDAR encephalitis as a disease model. Cutting-edge antibody engineering will define the unexplored contribution of antibody glycosylation (which modulates innate functions) to neuropathology. ImmuBRAIN combines human studies with experimental models. Human studies will extensively profile patients’ NMDAR antibodies using a novel antibodyomics platform; characterize the compartmentalized antibody responses between brain and periphery using B-cell receptor repertoire analyses; define the distribution of brain innate infiltrates and quantify neuronal death using advanced microscopy. In vitro models will determine the effects of patients’ and glycan-modified antibodies on neurons using confocal microscopy and electrophysiology; explore the neuron-microglia interactions during trogocytosis using super-resolution and live cell imaging. Finally, a new mouse model of NMDAR encephalitis will serve to define the dynamics of antibody and innate responses over time and at different disease stages, and explore the therapeutic role of in vivo antibody glycan modifications in reversing the observed behavioural and molecular alterations. These findings will revolutionize the current understanding of innate immune mechanisms in AE and other inflammatory brain disorders, and will provide novel antibody-modulating therapies (e.g., glycan modifications) that will ultimately improve patients’ outcomes.

Drug discovery in mental health is undergoing a crisis. Characterizing a new generation of biomarkers for the different disorders seems the promising path towards developing new treatments. Biomarkers, including behavioral biomarkers, are quantitative observables that are altered and can be used for diagnosis, prognosis or treatment response monitoring. Importantly for preclinical research, these biomarkers should be translational and work across species, but experiments in rodents are largely based on paradigms that are disparate from those used in humans. Training rodents in more human-like tasks has a strong translational potential, but it has important shortcomings: it requires developing finely-controlled tasks involving long training periods, consuming lots of animals and ultimately inducing very elevated personnel costs. To confront this experimental bottleneck , neuroscience and pharmaceutical laboratories are trying to automatize their methods, but still most systems rely on human intervention, require animal manipulation and impose a high maintenance toll. Leveraging our experience building behavioral systems during our ERC-CoG-2015, we will design, build and distribute the Mouse Village (MV), a new fully-automated open platform in which mice self-trained 24/7 in different cognitive tasks while displaying complex social interactions without human intervention. The MV consists in a system of cages connected to an operant touchscreen chamber that can implement multiple cognitive tasks. We will build on a previously developed pilot MV by introducing a more modular and flexible design and validate the system using several standard social and cognitive tests. The MV will be shared to the community under and Open Software and Hardware licenses. By eliminating the experimental bottleneck imposed by behavioral experiments, the MV has the potential to accelerate the search for new biomarkers of brain disorders and have a substantial impact on mental health.

Intrahepatic cholangiocarcinoma (iCCA) is the second primary malignancy of the liver with few therapeutic options, late diagnosis, chemotherapy resistance and dismal prognosis. iCCA incidence and mortality increased worldwide in the last decades, linked to the rise of risk factors including non-alcoholic steatohepatitis. iCCA is a highly heterogeneous tumor characterized by a complex and dynamic tumor microenvironment (TME) where stromal cells coexist with cancer and cancer stem cells (CSC). CSCs are a heterogeneous subpopulation of cancer cells capable of tumor initiation, malignant growth and responsible for chemoresistance. The main objective of the “SteMiCCA” project is to identify and target the unexplored CSC-TME interactions in iCCA, aimed to better understand the complex biology and to identify new therapeutic targets for this deadly disease. To achieve this objective, an interdisciplinary approach including in vitro (spheroids and organoids from human and mouse) and in vivo (ICC-models, subcutaneous and xenograft tumor induction) systems; high throughput transcriptomics (scRNA-Seq and RNA-Seq) and bioinformatics will be employed. Considering my strong background and experience in iCCA and TME, and the host institution’s well-recognized expertise in stem cell biology and liver cell plasticity, this proposal envisions a successful reciprocal transfer of knowledge. Moreover, this fellowship will allow me to gain expertise in supervision, intellectual property management, research funding and proposal writing, with the main goal to acquire more competences and reach professional maturity to start my own research line in iCCA. This will also be a first step to strengthen my network and start contributing to European actions such as ESCALON PROJECT 825510 and EURO-CHOLANGIO-NET, created to boost multidisciplinary studies with interest in basic, translational and clinical research in the field of CCA.

Obesity and aging are two of the most important health challenges facing our societies in the 21st century. However, there are currently no effective therapeutic approaches to mitigate the growing obesity pandemic or to promote healthy aging. Therefore, it is essential to explore new research frontiers in order to develop more effective and safer strategies. Mitohormesis, the process by which induced mitochondrial stress can trigger beneficial health effects, has emerged as a promising area of study in biomedical research. However, its potential effectiveness against metabolic and aging-associated disorders in vertebrate organisms is poorly explored. We have generated a mouse model that exhibits enhanced mitohormesis in the vascular endothelial cells. In this proposal, we intend to conduct a proof of concept to assess whether triggering endothelial mitohormesis can serve as a novel approach for treating obesity and age-related health issues. To this aim, we will execute 4 work packages to determine: (a) the preventive and (b) curative effect of endothelial mitohormesis on obesity and associated metabolic alterations; (c) the impact of the promotion of endothelial mitohormesis on aging markers; (d) the identity of new circulating mitohormetic mediators with the potential to be pharmacological targets. These studies will be carried out by a research team with an extraordinary track-record in the fields of metabolism and aging. Our objective will be bringing the technology from TRL3 to TRL4 by evaluating the performance and feasibility of the technology in a controlled laboratory environment. The research team will closely work with the Tech Transfer Office to ensure the implementation of a successful IPR strategy. A therapy based on the controlled induction of endothelial mitohormesis could have a great impact on the quality of life of millions of patients, reduce the burden of chronic diseases on the health system and have important economic repercussions.

The tumor microenvironment (TME) plays an essential role in tumor evolution and response to therapy. Understanding the complex network comprising tumor and immune cells is critical for the success of cancer immunotherapy. CLL and MCL are two non-Hodgkin’s lymphomas characterized by accumulating CD5+ small and mature B lymphocytes in peripheral blood, bone marrow, lymph nodes, and extranodal sites. Despite having distinct basic pathogenic mechanisms, they share the cell of origin and molecular alterations, clinical features, and epidemiological characteristics. However, the influence of the crosstalk between tumor cells and the TME on the disease progression and treatment resistance remains poorly understood, hindering the development of novel therapeutic possibilities. This study aims to uncover the intricate interplay between the TME and neoplastic cells in CLL and MCL and exploit its therapeutic use. To achieve this, I will use single-cell RNA sequencing and spatial transcriptomics to analyze the composition of non-tumoral cells in tumor-involved tissues and at different time points of the disease. Next, I will use correlative analyses to identify the molecular and cellular interactions between the tumor cell (epi)genomic configuration and the TME, determining tumor evolution and response to therapy. Finally, I will validate the identified drivers of disease evolution and treatment resistance and examine their potential as a target for treatment using a 3D LN-derived culture system. The findings have the potential to provide a solid basis for establishing new models of patient risk stratification and identify potential therapeutic opportunities targeting interactions between tumor cells and their microenvironment. Conducting this project at the Molecular Pathology of Lymphoid Neoplasms group of Prof. Dr. Elias Campo will provide me an opportunity to acquire essential technical and soft skills vital for advancing my career in research.