The clinical translation of nanoparticle-based therapies over the last decade has been hampered by issues such as inefficient targeting and limited therapeutic effect. This poor translational outcome calls for deeper understanding of the biomechanics of cell-nanoparticle (cell-NP) interactions. Indeed, targeting mechanosensing-activated cell pathways is suitable for tuning cell fate and readdressing its functions, as mechanosensing components control the expression of genes involved in the cell’s migration, survival and resistance to drugs. Hippo pathway appears to be one of the most promising mechanobiology pathway, as it is involved in pathological diseases and tissue regeneration. This project aims to address the response of this pathway on cells upon interaction with nanoparticles. Indeed, tuning cell mechanosensing with nanoparticles is likely to hold great potentiality to control cell functionalities. The first objective will be the synthesis of nanoparticles of different size, shape and stiffness, using a silica scaffold coated with hyaluronic acid via metal-phenolic network assembly with exceptional physicochemical properties. The second objective consists in the application of Superresolution microscopy for studying cell-NP interactions with unprecedented detail and unveil the interaction/structure/ spatiotemporal localization of mechanosensing components related to the Hippo pathway (i.e. YAP, actin and focal adhesions) at molecular level. The third objective will be the deep analysis of the molecular biology and biochemistry of mechanosensing proteins (i.e. YAP, TAZ, RhoA and Rock), and their downstream effectors (i.e. TEAD and transcriptional factors) involved in the response to cell-NP interaction. The forth objective will pursue the analysis of these interactions using NenoVision technology (LiteScope), for measuring cell stiffness at the boundary of cell-NP contact with unique resolution.

Excessive inflammation underlies the development of inflammatory diseases, e.g. Rheumatoid arthritis, Crohn's Disease, inflammatory bowel disease, and cardiovascular disease - the most common cause of death in the world. Human physiology is controlled by defined molecular mechanisms, commonly controlled by specific target nerves and their associated reflexes which optimize function – for example molecular mechanisms regulating inflammation. Harnessing the power of the inherent nerve reflexes which are evolutionarily adapted to regulate specific molecular mechanisms holds promise to overcome many of the drawbacks intrinsic to current drug therapies. This approach, designated “bioelectronic medicine”, presents a radically improved way to both treat and better understand disease – with increased adaptability and precision. The vagus nerve, as shown by the team in this project and other groups, plays a key role in regulating key pro-inflammatory cytokines, pro-resolving mediators, and controls inflammation. However, specific targeting of the vagus nerve has required neurosurgical implantation of a pacemaker-like stimulator device with a control unit, a connecting cable, and an electrode implanted at depth. The requirement for highly specialized surgery significantly limits the usefulness and possibilities for implementation of nerve stimulation in treatment of inflammation. Surgery and device implantation also carries risks for permanent tissue damage, local inflammation, infection, and other potentially serious complications. TREATMENT will generate a precise, non-invasive nerve stimulator that uses Temporal Interference in an ultra-thin form factor (enabling patch-based applications) that will (i) target one nerve of interest and avoid off-target stimulation, (ii) avoid surgery, (iii) simplify effective use and (iv) enable bioelectronic medicine.

Epilepsy is a brain disorder characterized by sporadic debilitating seizures and declines in mental health. Available therapies suffer from drawbacks and are not suitable for all patients. The medical needs in epilepsy encapsulate those for clinically managing brain health in general, i.e., the need for precision, personalized, non-systemic and safer therapies. Potential innovative solutions are available from advances in technology but will require multidisciplinary teams which bridge between engineering and clinical neurology. The main objective of the EMUNITI project is to develop a breakthrough, non-invasive, personalized brain stimulation device for epilepsy patients, (i) as a diagnostic for seizure localization which can guide subsequent surgical resection; and (ii) as a therapy, akin to immunizing the patient against seizures. The device is based on temporal interference (TI) of electrical signals delivered from positions around the scalp. The project will be led by Adam Williamson, who is a former recipient of ERC Starting and Proof-of-Concept grants and has IP for the technology. His lab is at the interface between state-of-the art engineering and neurology, and is based in a major European centre for clinical research on epilepsy in Marseille, France. His lab has already demonstrated in a pilot study of epilepsy patients, that TI can block seizure-provoking brain activity. The project includes three work packages (WPs). WP1 focuses on optimizing TI stimulation at the engineering level and establishing a novel form of TI in multipolar TI (mTI). WP2 will evaluate mTI in epilepsy patients, for seizure localization and for therapeutic intervention with the potential to create brain seizure immunity through repeated mTI. WP3 administers the project and delivers the main outcome: a clinical prototype with closed-loop stimulation combining mTI and intelligent software. The successful outcome is anticipated to lead to other applications for TI in brain health.

Pathological extracellular matrix (ECM) remodelling underlies the onset and progression of most cardiac conditions. The biomechanical stress associated with ECM remodelling modifies the transcriptional and post-transcriptional landscape of cardiac cells. While the role of RNA homeostasis in the pathological heart is being appreciated, our understanding of the molecular processes governing this balance is limited. Post-transcriptional RNA modification facilitates quick adaptation to pathological conditions. Therefore, understanding how these events affect ECM remodelling has the potential to identify early therapeutic targets in treating cardiac diseases. RNA binding proteins (RBPs) have been recently shown to contribute to the onset and progression of these pathologies. Mutations in the cardiac-specific alternative splicing regulator RBM20 directly impact the splicing of the sarcomeric protein Titin and cause cardiac remodelling. Many pathogenic variants in RBM20 cause a hereditary form of cardiomyopathy and affect the nuclear import of RBM20 thereby causing splicing dysfunction and leading to the expression of the fetal isoform of Titin. Here we propose to leverage our experience with induced pluripotent stem cell (iPSC)-based models of cardiac diseases and ECM remodeling to challenge the hypothesis that RBM20 and its mutants display a specific mechanosensitive behavior in the heart. We hypothesize that, first, pathological ECM remodelling affects RBM20 subcellular localization thereby resulting in mis-splicing of RBM20 targets such as titin and further disrupting the cardiomyocyte function. Secondly, RBM20 variants affecting titin splicing and causing ECM remodelling may impact the subsecellular localization of other RBPs therefore further worsening the heart failure phenotype. We believe the research activities proposed will complement the aims of the CARDIOREPAIR project by providing novel insights into the molecular process of mechanosensitive splicing.