ERC FUNDED PROJECTS

Inflammatory diseases affect over 80 million people worldwide and accompany many diseases of industrialized countries, being the majority of them infection-free conditions. There are few efficient anti-inflammatory drugs to treat chronic inflammation and thus, there is an urgent need to validate novel targets. We now know that innate immunity is the main coordinator and driver of inflammation. Recently, we and others have shown that the activation of purinergic P2X7 receptors (P2X7R) in immune cells is a novel and increasingly validated pathway to initiate inflammation through the activation of the NLRP3 inflammasome and the release of IL-1β and IL-18 cytokines. However, how NLRP3 sense P2X7R activation is not fully understood. Furthermore, extracellular ATP, the physiological P2X7R agonist, is a crucial danger signal released by injured cells, and one of the most important mediators of infection-free inflammation. We have also identified novel signalling roles for P2X7R independent on the NLRP3 inflammasome, including the release of proteases or inflammatory lipids. Therefore, P2X7R has generated increasing interest as a therapeutic target in inflammatory diseases, being drug like P2X7R antagonist in clinical trials to treat inflammatory diseases. However, it is often questioned the functionality of P2X7R in vivo, where it is thought that extracellular ATP levels are below the threshold to activate P2X7R. The overall significance of this proposal relays to elucidate how extracellular ATP controls host-defence in vivo, ultimately depicting P2X7R signalling through and beyond inflammasome activation. We foresee that our results will generate a leading innovative knowledge about in vivo extracellular ATP signalling during the host response to infection and sterile danger.

1 794 948 €

Start date: 2014-09-01, End date: 2019-08-31

Genome maintenance, chromatin remodelling and transcription are tightly linked biological processes that are currently poorly understood and vastly unexplored. Nucleotide excision repair (NER) is a major DNA repair pathway that mammalian cells employ to maintain their genome intact and faithfully transmit it into their progeny. Besides cancer and aging, however, defects in NER give rise to developmental disorders whose clinical heterogeneity and varying severity can only insufficiently be explained by the DNA repair defect. Recent work reveals that NER factors play a role, in addition to DNA repair, in transcription and the three-dimensional organization of our genome. Indeed, NER factors are now known to function in the regulation of gene expression, the transcriptional reprogramming of pluripotent stem cells and the fine-tuning of growth hormones during mammalian development. In this regard, the non-random organization of our genome, chromatin and the process of transcription itself are expected to play paramount roles in how NER factors coordinate, prioritize and execute their distinct tasks during development and disease progression. At present, however, no solid evidence exists as to how NER is functionally involved in such complex processes, what are the NER-associated protein complexes and underlying gene networks or how NER factors operate within the complex chromatin architecture. This is primarily due to our difficulties in dissecting the diverse functional contributions of NER proteins in an intact organism. Here, we propose to use a unique series of knock-in, transgenic and NER progeroid mice to decode the functional role of NER in mammals, thus paving the way for understanding how genome maintenance pathways are connected to developmental defects and disease mechanisms in vivo.

1 995 000 €

Start date: 2016-01-01, End date: 2020-12-31

The development of alternative greener synthetic methods to transform renewable feedstocks into elaborated chemical structures mediated by solar light is a prerequisite for a future sustainable society. In this regard, this project entails the use of visible light as driving force and water as a source of hydrides for the synthesis of high-value chemicals. The project merges photoredox catalysis with 1st row transition coordination complexes catalysis to open a new avenue for greener selective catalytic reduction processes for organic substrates. The ground-breaking nature of the project is: A) Develop light-driven region- and/or enantioselective catalytic reductions using well-defined cobalt coordination complexes with aminopyridine ligands, initially developed for water reduction. Sterics, electronics and supramolecular interactions (apolar cavities and chiral pockets) will be studied to proper control of the selectivity in the reduction of i) C=E and C=C bonds and ii) in the C-C inter- and intramolecular reductive homo- or heterocouplings. B) Fundamental understanding of the light-driven cobalt catalysed reductions characterizing intermediates that are involved in the reactivity, kinetics and labelling studies as well as performing computational modelling of reaction mechanisms. The basic understanding of operative mechanisms will expedite a new methodology for electrophile-electrophile umpolung couplings. C) Enhance catalytic performance of the light-driven cobalt catalysed reductions by self-assembling of catalyst-photosensitizer into carbon based pi-conjugated materials through noncovalent supramolecular interactions. Likewise, it will allow electrode immobilization for electrocatalysed reductions using water as a source of protons and electrons. As a proof of concept, cobalt catalysts based on aminopyridine ligands have been shown highly active in the light-driven reduction of ketones and aldehydes to alcohols, using water as the source of hydrogen atom.

1 999 063 €

Start date: 2015-07-01, End date: 2020-06-30

In InanoMOF, we aim to develop frontier Supramolecular and Nanochemistry methodologies for the synthesis of a novel class of structures via controlled assembly of nanoscale metal-organic frameworks (nanoMOFs) and inorganic nanoparticles (INPs). These methods will embody the premise that “controlled object-by-object nano-assembly is a ground-breaking approach to explore for producing systems of higher complexity with advanced functions”. The resulting hybrid nanoMOF@INPs will marry the unique properties of INPs (magnetism of iron oxide NPs and optics of Au NPs) to the functional porosity of MOFs. The first part of InanoMOF encompasses the design, synthesis-assembly and characterisation of nanoMOF@INPs - advanced MOF-based sorbents that incorporate the functionality of the INPs used: magnetically controlled movement, in vivo detectability, enhanced biocompatibility and porosity, pollutant removal, or controlled sorption/delivery. The second part of InanoMOF entails studying the physicochemical properties of the synthesised nanoMOF@INPs and ascertaining their utility as drug-delivery/theranostic systems and as magnetic sorbents for pollutant removal. Specifically, we will study their stability in working media and determine their capacities for drug or pollutant sorption/delivery capacities. As proof-of-concept, we will study their toxicity in vitro and in vivo; enhancement of their in vitro therapeutic efficacy; and their capacity to remove pollutants (in real water and gasoline/diesel fuel samples) via magnetic assistance. In InanoMOF we will endeavour to establish the synthetic bases for controlling the spatial ordering of nanoMOF crystals, whether alone or combined with other nanomaterials (e.g. INPs, graphene, etc.). We are confident that our work will ultimately enable researchers to create MOF-based composites having cooperative and synergistic properties and functions for myriad applications (e.g. heterogeneous catalysis, sensing and separation).

1 942 665 €

Start date: 2014-04-01, End date: 2019-03-31

InOutBioLight aims to design multifunctional rubbers with enhanced mechanical, thermal, color-converting, and light-guiding features towards advanced biohybrid lighting and photovoltaic technologies. The latter are placed at the forefront of the EU efforts for low-cost production and efficient consumption of electricity, a critical issue for a sustainable development. In this context, the use of biomolecules as functional components in lighting and photovoltaic devices is still a challenge, as they quickly denature under storage and device operation conditions. This paradigm has changed using an innovative rubber-like material, in which the biofunctionality is long preserved. As a proof-of-concept, color down-converting rubbers based on fluorescent proteins were used to design the first biohybrid white light-emitting diode (bio-HWLED). To develop a new generation of biohybrid devices, InOutBioLight will address the following critical issues, namely i) the nature of the protein-matrix stabilization, ii) how to enhance the thermal/mechanical features, iii) how to design multifunctional rubbers, iv) how to mimic natural patterns for light-guiding, and v) how to expand the technological use of the rubber approach. To achieve these goals, InOutBioLight involves comprehensive spectroscopic, microscopic, and mechanical studies to investigate the protein-matrix interaction using new polymer matrices, additives, and protein-based nanoparticles. In addition, the mechanical, thermal, and light-coupling features will be enhanced using structural biocompounds and reproducing biomorphic patterns. As such, InOutBioLight offers three major advances: i) a thorough scientific basis for the rubber approach, ii) a significant thrust of the emerging bio-HWLEDs, and iii) innovative breakthroughs beyond state-of-the-art biohybrid solar cells.

1 999 188 €

Start date: 2020-01-01, End date: 2024-12-31

Cardiac toxicity is one of the most frequent serious side effects of cancer therapy, affecting up to 30% of treated patients. Cancer treatment-induced cardiotoxicity (CTiCT) can result in severe heart failure. The trade-off between cancer and chronic heart failure is an immense personal burden with physical and psychological consequences. Current therapies for CTiCT are suboptimal, featuring poor early detection algorithms and nonspecific heart failure treatments. Based on our recently published results and additional preliminary data presented here, we propose that CTiCT is associated with altered mitochondrial dynamics, triggering a cardiomyocyte metabolic reprogramming. MATRIX represents a holistic approach to tackling mitochondrial dysfunction in CTiCT. Our hypothesis is that reverting metabolic reprogramming by shifting mitochondrial substrate utilization could represent a new paradigm in the treatment of early-stage CTiCT. By refining a novel imaging-based algorithm recently developed in our group, we will achieve very early detection of myocardial damage in patients treated with commonly prescribed cancer therapies, long before clinically used parameters become abnormal. Such early detection, not available currently, is crucial for implementation of early therapies. We also hypothesize that in end-stage CTiCT, mitochondrial dysfunction has passed a no-return point, and the failing heart will only be rescued by a strategy to replenish the myocardium with fresh healthy mitochondria. This will be achieved with a radical new therapeutic option: in-vivo mitochondrial transplantation. The MATRIX project has broad translational potential, including a new therapeutic approach to a clinically relevant condition, the development of technology for early diagnosis, and advances in knowledge of basic disease mechanisms.

1 999 375 €

Start date: 2019-09-01, End date: 2024-08-31

Nutrient-sensing by POMC neurons is a critical process to monitor the metabolic status of the organism and to coordinate adaptive neuroendocrine, behavioural and metabolic effectors of energy balance. Mitochondria, as central commanders of cellular energy production and primary sources of bioenergetic signals, are logical candidates to play a key role in metabolic control. However, a comprehensive understanding of the mitochondria as nutrient-sensors and modulators of systemic energy homeostasis is lacking. MITOSENSING hypothesizes that dedicated mitochondrial networks in POMC neurons are able to sense, integrate and respond to alterations in the nutritional milieu and engage physiological actions to maintain energy balance. Thus, defects in these mitochondrial nutrient-sensing programs in this subset of neurons underlie the development of metabolic conditions such as obesity and type-2 diabetes (T2D). To test it, we will pursue three aims: 1) to identify transcriptionally-modulated mitochondrial nutrient-sensing programs in POMC neurons; 2) to investigate whether disruption of specific nutrient-sensing programs in POMC neurons cause metabolic disorders; 3) to investigate whether the development of lifestyle-associated metabolic disorders are caused by defective mitochondrial nutrient-sensing programs in POMC neurons. By providing neuron-specific, integrative, functional and mechanistic in vivo strategies, MITOSENSING will represent a major step forward into the understanding of mitochondria as a nutrient-sensing entity, the gene programs involved and their physiological regulatory functions in the context of energy balance control. Adequate coordination of neuronal nutrient-sensing with energy balance control is critical to sustain life, and thus understanding the molecular mechanisms governing these physiological programs will have an enormous scientific impact and also potential therapeutical implications for obesity and T2D.

1 999 573 €

Start date: 2017-04-01, End date: 2022-03-31

Deregulation of the Myc oncogene promotes tumorigenesis in most if not all cancers and is often associated with poor prognosis. However, targeting Myc has long been considered impossible because of potential catastrophic side effects in normal tissues. Despite this general assumption, we showed that Myc inhibition displays extraordinary therapeutic benefit in various transgenic mouse models of cancer, and caused only mild, well-tolerated and reversible side effects in normal tissues. For these studies we have employed a dominant negative of Myc, called Omomyc, which we designed and validated, and that is able to inhibit Myc transactivation function both in vitro and in vivo. Omomyc has so far been utilized exclusively as gene therapy and served the purpose of pre-clinically validating the therapeutic impact of systemic Myc inhibition. In this proposal we intend to push such a therapeutic approach further towards the clinic, making use of 1. Omomyc-based Cell Penetrating Peptides (CPPs): a novel, state-of-the-art potential method for directly utilising Omomyc itself (or a similar peptide) as a drug; 2. A new generation of Myc inhibitory small molecules generated by our collaborators at the Roswell Park Cancer Center. Our study in different mouse models of cancer will provide a comprehensive preclinical validation of such innovative therapies and will potentially boost our therapeutic arsenal against the majority of human cancers

1 730 700 €

Start date: 2014-05-01, End date: 2019-04-30

In neurons, sites of Ca2+ influx and Ca2+ sensors are located within 20-50 nm, in subcellular “Ca2+ nanodomains”. Such tight coupling is crucial for the functional properties of synapses and neuronal excitability. Two key players act together in nanodomains, coupling Ca2+ signal to membrane potential: the voltage-dependent Ca2+ channels (VDCC) and the large conductance Ca2+ and voltage-gated K+ channels (BK). BK channels are characterized by synergistic activation by Ca2+ and membrane depolarization, but the complex molecular mechanism underlying channel function is not adequately understood. Information about the pore region, voltage sensing domain or isolated intracellular domains has been obtained separately using electrophysiology, biochemistry and crystallography. Nevertheless, the specialized behavior of this channel must be studied in the whole protein complex at the membrane in order to determine the complete range of structures and movements critical to its in vivo function. Using a combination of genetics, electrophysiology and spectroscopy, our group has measured for the first time the structural rearrangements accompanying whole BK channel activation at the membrane. From this unique position, our first goal is to fully determine the real time structural dynamics underlying the molecular coupling of Ca2+, voltage and activation of BK channels in the membrane environment, its regulation by accessory subunits and channel effectors. BK subcellular localization and role in Ca2+ nanodomains make these channels perfect candidates as reporters of local changes in [Ca2+] restricted to specific nanodomains close to the neuronal membrane. In our laboratory we have created fluorescent variants of the channel that report BK activity induced by Ca2+ binding, or Ca2+ binding and voltage. Our second aim in this proposal is to optimize and deploy this novel optoelectrical reporters to study physiologically relevant Ca2+-induced processes both in cellular and animal mode

1 999 742 €

Start date: 2015-09-01, End date: 2020-08-31

Finding the mutations, genes and pathways directly involved in cancer is of paramount importance to understand the mechanisms of tumour development and devise therapeutic strategies to overcome the disease. Due to their role in cancer development and maintenance, the proteins encoded by cancer genes are candidate therapeutic targets. Indeed, in recent years we have witnessed the development of successful cancer-targeting therapies to counteract the effect of driver mutations. Although the coding part of the human genome has now largely been explored in the search for cancer driver mutations in most frequent cancer types, the extent of involvement of noncoding mutations in cancer development remains a mystery. The main challenges faced are: 1) the functional role of most noncoding regions is unknown, and 2) tumours often have thousands of somatic mutations, so that distinguishing cancer driver mutations from bystanders is like finding the proverbial needle in a haystack. To overcome these two challenges I propose to analyse the pattern of somatic mutations across thousands of tumours in noncoding regions to identify signals of positive selection. These signals are an indication that mutations in the region have been positively selected during tumour evolution and are thus directly involved in the tumour phenotype. The large scale analysis proposed here will allow us to create a catalogue of noncoding elements involved in different types of cancer upon mutations. We will study in detail a selected set of driver elements to uncover their specific function and role in the tumourigenic process. Furthermore, we will explore possibilities of counteracting their driver effect with targeted drugs. The results of this project may boost our understanding of the biological role of noncoding regions, help to unravel novel molecular causes of cancer and provide novel targeted therapeutic opportunities for cancer patients.

1 995 829 €

Start date: 2016-12-01, End date: 2021-11-30

Just as our experience has its origin in our perceptions, our perceptions are fundamentally shaped by our experience. How does the brain build expectations from experience and how do expectations impact perception? In a Bayesian framework, expectations determine the environment’s prior probability, which combined with stimulus information, can yield optimal decisions. While the accumulation-to-bound model describes temporal integration of sensory inputs and their combination with the prior, we still lack electrophysiological evidence showing neural circuits that integrate previous events adaptively to generate advantageous expectations. I aim to understand (1) how circuits in the cerebral cortex integrate the recent history of stimuli and rewards to generate expectations, (2) how expectations are combined with sensory input across the processing hierarchy to bias decisions and (3) whether the dynamics of the expectation can dominate neuronal and choice variability. I will train rats in a new auditory discrimination task using predictable stimulus sequences that, once learned, are used to compute adaptive priors that improve discrimination. I will perform population recordings and optogenetic manipulations to identify the brain areas involved in the computation of priors in the task. To reveal the circuit mechanisms underlying the observed dynamics I will train a computational network model to classify fluctuating inputs and, by adapting its dynamics to the statistics of the stimulus sequence, accumulate evidence across trials to maximize performance. The model will generalize the accumulation-to-bound model by integrating information across various time scales and will partition choice variability into that caused by the dynamics of the prior or by fluctuations in the stimulus response. My proposal points at a paradigm shift from viewing neuronal variability as a corrupting source of noise to the result of our brain’s inevitable tendency to predict the future.

2 000 000 €

Start date: 2016-09-01, End date: 2021-08-31

The precise synthesis of nano-devices with tailored complex structures and properties is a requisite for their use in nanotechnology and medicine. Nowadays, the technology for the generation of these devices lacks the precision to determine their properties, and is accomplished mostly by “trial and error” experimental approaches. Bottom-up self-assembly that relies on highly specific biomolecular interactions of small and simple components, is an attractive approach for nanostructure templating. Here, we propose to overcome aforementioned challenges by using self-assembling protein building blocks as templates for nanofabrication. In nature, protein assemblies govern sophisticated structures and functions, which are inspiration to engineer novel assemblies by exploiting the same set of tools and interactions to create nanostructures with numerous potential applications in synthetic biology and nanotechnology. We hypothesize that we can rationally assemble a variety functional nanostructures by the logical combination of simple protein building blocks with specified properties. We propose to use a designed repeat protein scaffold for which we acquired a deep understanding of its molecular structure, stability, function, and inherent assembly properties. Only few conserved residues define the structure of the building block, which allow us to mutate its sequence to modulate assembly properties and to introduce reactive functionalities without compromising the structure of the scaffolding molecule. First, we will design a collection of protein-based nanostructures. Then, we will introduce reactive functionalities to create hybrid nanostructures with nanoparticles, metals and electro-active molecules. Finally, these conjugates will be used to build nano-devices such as nanocircuits, catalysts and electroactive materials. The outcome of this project will be a modular versatile platform for the fabrication of multiple protein-based hybrid functional nanostructures.

1 718 850 €

Start date: 2016-01-01, End date: 2020-12-31

Our memory system is optimised for remembering the exceptional over the mundane. We remember better those events that violate predictions generated by the prevailing context, particularly because of surprise or emotional impact. Understanding how we form and retrieve long-term memories for important or salient events is critical for combating the rapidly growing incidence of pathologies associated with memory dysfunction with huge socio-econonomic burden. Human lesion and non-invasive functional imaging data, motivated by findings from animal models, have identified subcortical structures that are critical for upregulating hippocampal function during salient event memory. However, mechanistic understanding of these processes in humans remains scarce, and requires better experimental approaches such as direct intracranial recordings from, and focal electrical stimulation of, these subcortical structures. This project will characterise human subcortico-cortical neuronal circuit dynamics associated with enhanced episodic memory for salient stimuli by studying direct recordings from human hippocampus, amygdala, nucleus accumbens, ventral midbrain and cortex. Within this framework, I will elucidate the electrophysiological mechanisms underlying amygdala-hippocampal-cortical coupling that lead to better memory for emotional stimuli, extend the hippocampal role in detecting unpredicted stimuli to define its role in orchestrating cortical dynamics in unpredictable contexts, and discover the neuronal response profile of the human mesolimbic dopamine system during salient stimulus encoding. The predicted results, based on my own preliminary data, will offer several conceptual breakthroughs, particularly regarding hippocampal function and the role of dopaminergic ventral midbrain in memory. The knowledge gained from this project is a fundamental requirement for designing therapeutic interventions for patients with memory deficits and other neuropsychiatric disorders.

2 000 000 €

Start date: 2019-05-01, End date: 2024-04-30

The S-CAGE project aims to develop a new generation of crystalline solids with periodically-organized discrete voids, or compartments, that would benefit from the combination of the high stability and robustness of dense materials with the structural diversity and versatility (and therefore large applicability) of open frameworks. These coordination polymers (CPs) will be capable of interacting with guest species in the absence of large channels or permanent pores due to the presence of dynamic entrances. This could open new horizons towards the design of unprecedented materials as an enhanced interplay between the guests and the frameworks will be achieved resulting from the confined space of the compartmentalized pockets. The main goals of S-CAGE will be: i) Chemical design of compartmentalized 1D, 2D and 3D coordination polymers. These materials will be designed in such a way that they will provide ideal room to accommodate different guest molecules, which can be easily tuned depending on the target guest. ii) Advanced structural characterization, including modern diffraction studies under pressure of gas and volatile guests. This strategy will provide unequivocal prove of the location of the guest molecules in the internal voids and gain insights of the mechanism of entrance. The direct visualization of the modes of interactions of different gases will permit a deep comprehension of the nature of their interaction. iii) Gas separation studies. My goal will be the development of materials that could specially serve for gas separation and improve the performances of zeolites and MOFs by implementation of dynamic entities into the framework. iv) Sensing capabilities through changes in magnetic properties. The chemical design followed in S-CAGE will result in magnetic CPs with confined spaces which should enhance the interaction of the guest molecules with the framework, and thus a change in their magnetism is expected.

1 998 750 €

Start date: 2017-05-01, End date: 2022-04-30

Alzheimer´s disease (AD) is the most common form of dementia, with over 35 million people suffering from it worldwide, and it constitutes a personal and societal tragedy of immense proportions. Fifty years of intense research have revealed many key elements of the biology of this neurodegenerative disorder. However, our understanding of the molecular bases of the disease is still very limited, and the available medical treatments for AD are purely symptomatic and hardly effective. It is now clear that the modulation of a single target is unlikely to yield the desired outcome, and we should move from gene-centric to network-centric therapeutic strategies. In addition, we should focus on early (asymptomatic) phases of AD, before the brain damage is irreversible, and the identification of molecular biomarkers to monitor the response of patients is paramount. Accordingly, the main objective of our proposal is the identification of novel biomarkers in AD to monitor the onset and progression of the pathology from very early stages, and to discover combinations of drug targets and chemical compounds able to modify the biology of the disease. We will first run proteomics and transcriptomics experiments, in AD mouse models, to reveal the organization of proteins and genes that are up- or down-regulated at different ages and AD stages, and their potential translocation into/out of mitochondria. We will then construct the AD-associated network, incorporating clinical data, which we will use as a framework for the integration and analyses of the –omics data collected. We will transform the static data snapshots, corresponding to the different AD stages, into a dynamic model able to explain the progression of the disease, providing hints as to the best strategies to monitor and modulate AD evolution. We will finally design and validate a systems pharmacology strategy, based on concerted multi-target perturbations with small molecules, to modify the biology of the disease.

1 296 000 €

Start date: 2014-04-01, End date: 2019-03-31

Throughout development and adult life, the growth and remodelling of living tissues is determined by a complex interplay between chemical and physical cues. Among such physical cues, mechanical tension is emerging as central regulator of cellular fate and function. To explain tension regulation, current research emphasizes molecular mechanisms at an ever increasing level of detail. How these local mechanisms are integrated to give rise to global patterns of tissue tension is unknown, however. The goal of this project is to provide a multiscale understanding of tension regulation within epithelial cell monolayers. We propose two pairs of competing mechanisms for tension regulation: fluidization vs. reinforcement and cell division vs. apoptosis. These competing mechanisms are inherently multiscale in the sense that they span and couple multiple levels of tissue organization, from the local actomyosin contraction to the multicellular rearrangement. For each of these competing mechanisms we plan to study how local processes and interactions give rise to global tensional patterns. To do this, we propose to develop an integrated experimental setup to map and perturb monolayer tension at different length scales. We expect this project to unveil the repertoire of mechanisms that epithelial tissues use to regulate their tension and dynamics.

1 981 761 €

Start date: 2015-01-01, End date: 2019-12-31

DNA topoisomerases are conserved nuclear enzymes that regulate DNA topology by transiently cleaving and resealing the DNA molecule, fulfilling a fundamental role in virtually every aspect of chromosome metabolism. Nevertheless, erroneous or abortive topoisomerase activity can result in persistent DNA strand breaks with the enzyme covalently attached to 3’ or 5’ DNA ends by a phosphotyrosyl bond, an anomalous structure that can compromise cell survival and/or genome integrity with the consequent implications in tumourigenesis. This peculiarity of topoisomerase catalysis also underlies the anticancer efficacy of topoisomerase poisons, which inhibit the re-ligation step of the reaction inducing the formation of DNA breaks that preferentially target highly proliferating and/or repair defective tumour cells. In addition to this link with cancer therapy, defects in the repair of topoisomerase-induced DNA damage have been linked to neurological disease. Understanding the cellular response to topoisomerase-induced breaks is therefore key for important aspects of human health, with possible implications in the development of novel diagnostic, prognostic and therapeutic tools. This project aims at acquiring a comprehensive picture of the dynamics of topoisomerase-induced DNA breaks: from their occurrence and repair to the consequences for genome expression and integrity. We rely on the development of completely novel assays to detect and isolate the different intermediates of topoisomerase-induced break repair, and which overcome major traditional limitations in the field. These tools are subsequently used to integrate the time-dependent and genome-wide distribution of the different steps and final outcomes of the process of topoisomerase-induced DNA break repair. Furthermore, we outline original proteomic and genetic screenings to identify novel factors and pathways specifically involved the cellular response to this important type of DNA lesion.

2 000 000 €

Start date: 2015-11-01, End date: 2020-10-31

The overreaching goal of my lab is to improve the prognosis of patients with high-risk pediatric brain tumors. To this end, I propose to integrate clinical and lab-based research to develop tumor-targeted oncolytic adenoviruses with the capacity to elicit a therapeutic immune response in those tumors. Our research will use novel and relevant models to accomplish the experimental aims. We have previously worked with Delta-24-RGD (DNX-2401) a replication-competent adenovirus that has been translated to the clinical scenario. In 2017, the first clinical trial phase I with DNX-2401 for newly diagnosed Diffuse Intrinsic Pontine Gliomas (DIPG; a lethal pediatric brain tumor) opened propelled by my team. Preliminary results from the first trials revealed that the intratumoral injection of the virus instigated an initial phase of oncolysis followed by a delayed inflammatory response that ultimately resulted in complete regression in a subset of the patients without associated toxicities. I hypothesized that enhancement of the immune component of the DNX-2401-based therapy will result in the complete regression of the vast majority of pediatric brain tumors. In our specific approach, we propose to understand the immune microenvironment of DIPGs and the response to viral therapy in the context of the trial. Moreover, that knowledge will leverage the design of Delta-24-based adenoviruses to recruit lymphocytes to the tumor with the competence of different type of ligands to activate the tumor infiltrating lymphocytes. I expect that this combinatorial innovative treatment will efficiently challenge the profound and inherent tumor immunosuppression and, in turn, will elicit a robust anti-tumor immune response resulting in the significant improvement of the prognosis and quality of life of patients with pediatric brain tumors. This project has the potential to produce a vertical advance in the field of pediatric oncology.

2 000 000 €

Start date: 2019-03-01, End date: 2024-02-29