Spinal muscular atrophy (SMA) is a fatal neurodegenerative disorder in children, characterized by motor neuron loss and atrophy of muscle. However, SMA affects many organ systems, since the causal gene, SMN1, is expressed in all cells. Recently, an AAV9-mediated gene therapy, Zolgensma, was approved. However, challenges remain. To be effective, the systemically delivered AAV9- SMN1 needs to mimic the native regulation of the genomic SMN1. In fact, recent findings demonstrated that delivery of AAV-SMN1 under the control of the ubiquitous viral promoter CMV found in Zolgensma eventually caused death of neurons in a mouse model of SMA. To date, there has been no systematic categorization of how treatment with SMN1 is regulated in different tissues through development. Furthermore, little is understood about the chromatin changes that occur during SMA, how gene therapy impacts the long-term stability of patient’s epigenomes, or how the AAV-delivered DNA is epigenetically controlled over time. The goal of the proposed work is to understand how the epigenome reacts to gene therapy, using SMA as a model. To do so I will: i) profile the epigenetic changes during SMA disease progression in mice, before and after gene therapy, ii) evaluate the role of various promoter-enhancer structures in the vector on the epigenetic regulation of AAV-delivered episomal DNA and iii) test if endogenous promoter-enhancer combinations can be used to achieved a better tuning of the delivered gene. The proposed project combines the expertise of Dr. Piera Smeriglio in SMA models, neuromuscular disorders and gene therapy with my expertise in epigenetic profiling, gene regulation, functional genomics, and single-cell technologies. AAV gene therapies are poised to have a huge impact on rare diseases, a key priority of the Horizon Europe programme, making now a critical time to understand their effect on the epigenome and cell regulatory networks to increase the efficacy and safety of their use.

Sarcopenia, marked by the progressive loss of muscle mass and strength due to aging, presents a significant public health challenge. Despite its increasing prevalence, treatment options remain limited, focusing primarily on exercise and nutraceuticals. Sarcopenia shares several pathological features with nerve injury-induced muscle atrophy, including neuromuscular junction (NMJ) degeneration. Recent studies have shown that muscle-resident Plp1+ glial cells rapidly activate a neurotrophic program in response to nerve damage. However, despite their established role in nerve regeneration, the involvement of these cells in age-related NMJ loss and muscle atrophy remains largely unexplored. The GLI-AGE project aims to explore the potential involvement of glial cells in age-related sarcopenia. It will utilize HD Spatial Gene Expression technology (HD-ST), a cutting-edge technique that combines advanced sequencing and high-resolution imaging to map glial cell activity and interactions in aged and geriatric mice. The project’s main objectives are: (1) to spatially characterize glial cell networks in aged muscle compared to young muscle, and (2) to validate their regenerative potential by identifying molecular targets (in-vivo and in-vitro) that may be implicated in the progression of sarcopenia. Through its multidisciplinary approach, GLI-AGE seeks to uncover novel therapeutic targets to preserve neuromuscular function and slow muscle degeneration in the elderly. The project’s findings could pave the way for innovative strategies to maintain muscle health, reduce frailty, and enhance the quality of life for older adults, aligning with Horizon Europe’s goal of promoting healthy and active aging.

THERABOT pioneers a groundbreaking approach in the fight against cancer through the development of engineered biohybrid agents, iBots. These iBots represent a significant leap in bacterial cancer therapy (BCT), combining synthetic biology and nanotechnology to encapsulate therapeutic bacteria with protective polymeric coatings. This encapsulation aims to enhance the delivery and efficacy of bacteria to colonize tumor tissue, drastically reducing the adverse immune responses and systemic toxicity associated with traditional BCT. THERABOT's innovative strategy focuses on colorectal cancer, employing a multifaceted approach that includes the design of advanced intelligent biohybrids by programming iBots for targeted therapeutic action, and validating their effectiveness in vivo. Central to THERABOT's mission is its interdisciplinary and international consortium, comprising 12 members from 10 leading academic institutions and 2 SMEs across Europe (France, Spain and Sweden) and from non-EU countries (Japan, Argentina, Chile, Brazil and Thailand). This collaboration fosters a unique integration of diverse scientific disciplines, including bioengineering, immunology, material science and oncology, promoting the exchange of knowledge, techniques, and cultural perspectives, which is pivotal for addressing the complex challenges of developing a new cancer therapy. THERABOT not only emphasizes the scientific innovation but also prioritizes the training and career development of seconded staff, enhancing their research competencies and preparing them for future challenges in diverse fields. THERABOT's focus on developing accessible and patient-friendly cancer treatments, underscores its alignment with EU priorities in health and innovation. By redefining cancer treatment paradigms towards more precise, effective, and personalized approaches, our project aspires to have a lasting impact on society, improving patient outcomes, and contributing to the global fight against cancer.

The objective of this network is to train, through research, a group of 10 young scientists, giving them the practical skills and knowledge necessary to tackleThe function of a neuron is determined by its expression of morphogen receptors and adhesion proteins (to pattern its connectome), of synaptic protein complexes (for intercellular communication), and of ion channels (for neuronal excitability and signal propagation). The protein composition of any neuron is itself dependent on gene expression controlled by transcription factors and by post-transcriptional regulations (splicing, translation, protein stability). Finally, neurons are highly compartmentalized cells and require trafficking of proteins to their correct locations. Alterations of these neuronal features can therefore be observed at multiple levels: molecular, cellular, and behavioral. We will focus on three neuronal features: i) gene expression levels as measured by RNA sequencing and proteomics, ii) assembly of functional multiprotein complexes (by high resolution microscopy and proximity dependent protein labelling), and iii) phenotypic output via a combination of optogenetics, calcium imaging, and behavioral assays. This research program will help define the critical features that differentiate between physiological and pathological states of a given neuron. Such biomarkers would be precious to facilitate drug screens or develop novel therapeutic strategies for human neurological diseases. The interdisciplinary research and training programme will cover a broad spectrum of approaches and topics, from intracellular molecular interactions to cell–cell communication, gene expression profiling, the impact of deleterious mutations, behavioral studies, and translation regulation. Coupled with extensive network-wide transferable skills training, this will prepare the fellows for careers in the medically important field of neurobiology as well as a broad range of carriers including government and industry.

Rare diseases, though individually infrequent, collectively pose important challenges for patients, clinicians and researchers in terms of diagnostics, healthcare and treatment. Myotonic Dystrophy (DM), the most predominant inherited muscular dystrophy in adults, impacts 60,000-70,000 individuals in Europe. Its complex genetic and clinical variability result in a lack of robust genotype-phenotype correlations, which complicates the understanding of tissue-specific disease mechanisms, the development of effective therapies and the stratification of patients into well-defined clinical groups for clinical trial purposes. In recent years, Antisense Oligonucleotides (ASO) have emerged as promising therapeutics, notably in neuromuscular disorders. However, previous clinical trials in DM have failed due the reduced efficacy and bioavailability of the ASO tested. ENTRY-DM aims to train 14 DCs in translational research, through the combination of basic and clinical competencies across multiple disciplines, as well as strong soft and transferable skills. Our interdisciplinary network seeks to enhance scientific and technological knowledge, spanning from disease mechanisms, ASO design and delivery strategies, to clinical trial preparedness, using innovative multidisciplinary approaches and best practices. The consortium includes experts in DM research, bioengineering of model systems for preclinical drug screening, ASO chemistry, as well as clinical and neuropsychological assessment. Close collaborations with multi-sectoral partners will address the challenges in technology transfer, providing high-quality bench-to-bedside training to the next generation of researchers. Through these efforts, ENTRY-DM will catalyze ASO therapeutic development towards upcoming impeding clinical trials, establishing solid foundations for future clinical applications, increased investment and entrepreneurship ventures in the field of DM and other related diseases.