Ischemia-reperfusion injury is a major cause of morbidity and mortality transversal to different clinical settings and pathologies, including myocardial infarction. Ischemia is characterised by insufficient supply of blood to tissues causing tissue oxygen deprivation. Importantly, damage by prolonged ischemia is further aggravated by reperfusion, leading to irreversible tissue damage. Identifying and understanding the factors contributing to ischemia-reperfusion injury is essential to counteract the irreversible damage and to potentially develop novel therapeutic approaches to intervene in myocardial infarction. Recent studies have shown that microRNAs control various aspects of heart disease, including ischemia-reperfusion injury. Although a few microRNAs have already been implicated in this process, a comprehensive analysis of the functional role of microRNAs in cardiac ischemia-reperfusion injury is still missing. The main goal of this project is to identify microRNAs controlling the resistance/susceptibility of cardiomyocytes to ischemia-reperfusion injury and characterise the molecular mechanisms underlying their function. This will be achieved through the combination of gain- and loss-of-function high-throughput screenings using genome-wide microRNA libraries, and deep-sequencing analysis of microRNA expression in conditions mimicking ischemia and ischemia-reperfusion in vitro and in vivo. The identification and characterisation of the molecular targets of the selected microRNAs will entail a combination of computational and experimental approaches This project uses innovative experimental approaches and will unravel a previously unappreciated network of microRNAs and microRNA targets critical to ischemia-reperfusion injury, which may reveal novel opportunities for therapeutic intervention, with important clinical implications.

Staphylococcus aureus is an opportunistic bacterial pathogen that causes acute and chronic infections responsible for a broad spectrum of human diseases. The high prevalence of antibiotic-resistant strains in clinical context and the spreading of these strains in the community, make S. aureus infections a leading cause of morbidity and mortality in developed countries. A better understanding of the factors contributing to bacteria resistance and host-pathogen interaction is essential to counteract S. aureus infection and, ultimately, to develop more effective therapeutic approaches. MicroRNAs (miRNAs) are genome-encoded small non-coding RNAs, which play a pervasive role in the post-transcriptional control of eukaryotic gene expression. Emerging evidence supports a determinant role for miRNAs in the interplay between bacterial pathogens and host cells, as part of the immune response, or as a molecular strategy exploited by pathogens to subvert host cellular functions. However, a comprehensive analysis of host miRNAs function during infection by S. aureus is still missing. The main goal of this project is to identify and characterize host miRNAs, which are critical for S. aureus-host interaction at the cellular level. This will be achieved through interdisciplinary and innovative strategies, such as large-scale genome-wide miRNAs screenings and high-content analysis. To increase the biological relevance of the study, the effect of selected miRNAs will be validated in S. aureus clinical isolates obtained from osteomyelitis patients. The identification and characterization of miRNA molecular targets will be based on computational and experimental approaches. Through the application of cutting-edge techniques, this project will reveal previously unappreciated factors/pathways essential for S. aureus infection, providing insights into the complex host-pathogen interplay. Additionally, the identified novel factors/pathways may be explored for therapeutic intervention.

Therapeutic drug delivery to the brain remains an unsolved challenge that limits the treatment of neurological and brain diseases, such as Glioblastoma, Parkinson and stroke. The main limitation is in the ability of the therapeutic drugs to cross the Blood Brain Barrier (BBB). To overcome this limitation, scientists present hyperthermia as an approach to overcome this limitation. Although this approach show promising results, the studies were not focused on understanding and combining the temperature raise during the BBB opening. Thus, the possibility of using this temperature rise to both improve the BBB opening performance and to exploit it to treat brain diseases is not used. For the aforementioned to be actualized, this project will bring the latest advances in intracellular temperature and nano-thermometry to a deeper understanding of the role of temperature in the BBB opening and its possible utilization in the treatment of brain diseases, such as glioblastoma. Thus, with the effort of the host organization (world leader in develop of nanomedicine platforms) and the experienced researcher (expert in nano-thermometry), two important objectives will be achieved under this Marie Skłodowska-Curie fellowship: (1) detailed studies and understanding of both the intracellular temperature and extracellular temperature needed for the BBB opening and (2) design and synthesis of the nano-complex for simultaneously BBB opening and the release “in real time and on demand” of therapeutic drugs for glioblastoma treatment in vivo. The outcome of this proposal will represent an important step forward in the field of brain drug delivery, that will not only improve the applications of nanotechnology in glioblastoma treatment, but will also facilitate the overcoming of the actual limitations in the treatment of different brain diseases.

Neurological disorders are a global leading medical and societal challenge, affecting particularly the aged population. Because of the lack of efficiency of pharmacological options, alternatives such as electrical stimulation using implanted electrodes has gained popularity. However, this strategy lacks spatiotemporal control over the stimulation of specific neural circuits, of which no biochemical information can be acquired. Recent breakthroughs in non-genetic, light-mediated electrical stimulation have shown promising results towards minimally invasive devices with controllable performance following exposure to near infrared (NIR) light. The LIGHTEST project proposes an innovative neural interface constituted by electrodes combining graphene with upconversion nanoparticles (UCNP), which will convert NIR light into electricity with spatiotemporal control. We will use novel tools to screen formulated graphene-UCNP complexes and evaluate their biological interactions in the presence of NIR light (e.g. high-throughput screening, RNA sequencing). This project counts on the applicant’s knowledge in synthesis, characterisation and biological evaluation of graphene-based materials, alongside the expertise of Dr Lino Ferreira in the formulation of light-responsive nanomaterials and their in vivo interactions, in combination with the experience in optogenetic engineering and mechanistic understanding of light-mediated stimulation in the brain from Dr João Peça at the Host institute. This project will contribute to the establishment of the applicant in Portugal as an independent researcher in nanomedicine, which is a strategic field that will boost European competitiveness in the development of novel therapies and inspire new generations for a better healthcare.