Project Portugal 2030

Summary

FFI is an ultra-rare genetic prion disease caused by highly penetrant mutations of the prion protein (PRNP) gene at the codon 178. Clinically, it is characterized by progressive insomnia, with autonomic and motor hyperactivation. The age of onset varies between 36 and 62 years of age, with a fatal outcome after 8 to 72 months. In terms of neuropathophysiology, FFI patients have a severe bilateral loss of neurons, with reactive gliosis (8,9). Currently, there is no cure for this disorder and much is unknown about the mechanisms of this disease. So, diagnosing FFI poses significant challenges due to its rarity and the variability of symptoms. It often requires a combination of clinical evaluation, genetic testing to identify the PRNP mutation, and sometimes specialized tests (e.g. polysomnography). Despite ongoing research efforts, there remains an urgent need to develop effective treatments for FFI, given its devastating impact on patients' physical and mental well-being. Hence, our goal is to deepen the knowledge of FFI by thoroughly characterizing the disease, developing a model for it, and investigating potential therapeutic approaches through: 1. Characterization of FFI Mutation Carriers: Clinical Evaluation: Conduct a comprehensive clinical assessment of FFI mutation carriers and respective controls, focusing on neurological manifestations and sleep parameters. Clock Genes Expression: Analyze the expression of clock genes in peripheral blood mononuclear cells (PBMCs) of FFI mutation carriers to understand their potential role in the pathogenesis of the disease. Lipidomic Evaluation: Assess the lipid profile using lipidomics to identify lipid alterations associated with FFI. 2. Establishment of a Human FFI Disease Model: Generation of Functional Hypothalamic Neuronal Cultures: Develop a clinically relevant model to study FFI pathophysiology by generating functional hypothalamic neuronal cultures derived from patient cells. Evaluation of Circadian Rhythms: Assess the circadian rhythms of the hypothalamic neurons by analyzing the expression of clock genes and monitoring sleep-related features using EEG-like measurements. Prion Protein Accumulation: Investigate prion protein accumulation and propagation within the neuronal cultures to elucidate the mechanisms underlying FFI pathology. Metabolic Profiling: Evaluate the metabolic profile of the in vitro model by quantifying lipid droplets content, providing insights into metabolic dysregulation associated with FFI. 3. Application of Therapeutic Techniques: CRISPR-Cas9 Gene Editing: Utilize CRISPR-Cas9 technology to correct the FFI gene mutation in patient-derived cells, aiming to develop a novel therapeutic approach for FFI. Circadian rhythm and prion-regulating treatment: Explore the use of rhythm-regulating drugs to prevent or inhibit prion accumulation, offering potential therapeutic strategies to mitigate FFI progression. Overall, this project represents a holistic, multidisciplinary effort that fosters a synergistic relationship between scientific breakthroughs and clinical applications. Moreover, the methodologies suggested introduce innovative techniques crucial for advancing human-centric therapeutic research.