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Protein misfolding is presently recognized as the underlying pathogenic mechanism on a diverse and growing number of severe human diseases (e.g. neurodegenerative diseases and specific cases of hereditary cancer) driving the development of new therapeutic approaches aimed at rescuing the folding of the target protein by adapting their cellular homeostasis (proteostasis). Therefore, in this curricular unit the students will acquire skills, at a theoretical, practical and laboratory level, that will allow them to: identify the intracellular protein quality control systems; understand the pathogenic impact of protein misfolding; design and apply experimental strategies to identify potential therapeutic targets within the proteostasis network and molecules with potential therapeutic effect (pharmacological proteostasis modulation); be familiar with novel technologies to study the cellular pathways involved in maintenance of a healthy proteome; learn how to manipulate scientific equipment and specialized software and how to analyze and present experimental data.
Theoretical Course
Proteins: Structure and Function; Folding in vitro (thermodynamic and kinetic bases); Biosynthesis and Protein Processing (folding in vitro/in vivo; the ribosome; co- and post-translation folding; folding in different cell compartments; the molecular chaperones and co-chaperones systems); Protein Degradation Pathways (half-lifetime; the N-degrons; the Ubiquitin-Proteasome and Autophagy-Lysosomal Systems; degradation in the different cell compartments); Proteotoxic stress response systems (UPR and HSR); Conformational diseases (loss-of-function and gain-of-function; genetic diseases and missense mutations; small molecules modulators of intracellular folding: pharmacological chaperones and proteostasis regulators); Orphan Drugs (rare diseases; orphan drugs designation; regulations and incentives; committee of orphan drugs; examples of orphan drugs).
Practical Course
Practical course will contribute to a theoretical/laboratory teaching interplay, acting not only as a support to the techniques that will be used in the laboratory but also extending the knowledge to techniques available for the study of proteostasis and for the identification and development of molecules with potential pharmacological effect, namely: laboratory tools for structural and functional protein characterization (in vitro and in cellulo methods); experimental approaches for the identification of pharmacological chaperones and proteostasis regulators (High-Throughput Screening Platforms; compound libraries; compound screenings: random/oriented and in vitro/in cellulo); Bioinformatic tools for the study of proteins and for predicting changes in protein stability (DG).
Laboratory Course
In the hands-on laboratory classes, students will execute techniques that will allow them to produce, purify and perform a comparative characterization of two recombinant human proteins (native wild-type form and a misfolded pathogenic variant). They will also use experimental methodologies that will allow them to monitor the stabilizing effect of potential therapeutic molecules (pharmacological chaperones).