CELLULAR MODELS AND NEUROEPIGENETICS

The research activity of the Unit deals with the preparation and characterization of cellular and tissue models for the study of the basic mechanisms of neurological/neurodegenerative diseases, such as amyotrophic lateral sclerosis and dementias. For this purpose, both primary cell lines (peripheral blood mononuclear cells and lymphoblastoid cell lines) and reprogrammed lines such as induced pluripotent stem cells (iPSCs), obtained from patients and controls, are used. IPSCs are subsequently differentiated into different neuronal and glial cell types. Currently, new three-dimensional cell models are being developed using the 3D bio-printer and, in particular, brain organoids are being generated for the study of neurodegenerative and neurodevelopmental diseases. These models are characterized by the use of basic techniques (immunofluorescence, real-time PCR and western blot) and innovative techniques (RNA-seq, single cell analysis and confocal microscopy).

The obtained cellular models are used for the study of some of the main pathogenetic mechanisms (mitochondria alterations and autophagic/mitophagic pathway, HDAC6 and HSP90) both for the study of epigenetic variations, i.e. of the heritable changes in gene expression which do not involve changes at the DNA sequence level. Regarding the epigenetic studies, the research activity focuses on the most frequent histone modifications, i.e. acetylation/deacetylation, methylation, ubiquitination, sumoylation, and phosphorylation by means cell biology and next generation ELISA techniques (AlphaLisa). In addition to the studies on histone modifications, the Unit also deals with DNA methylation studies by gene-specific pyrosequencing and the study of molecules that interact with G-quadruplexes, that are the basis of structural modifications of chromatin and transcriptomic variability.

Thanks to these different approaches it will be possible to clarify the main basic mechanisms and the epigenetic processes involved in neurodegenerative and neurodevelopmental diseases. This knowledge can be transferred to the clinical context to improve future therapeutic approaches.

These activities are carried out as part of research projects funded by various entities, such as the Ministero della Salute (Ricerca Corrente), the Cariplo Foundation, MIUR, ARISLA and Banking Foundations.