MOLECULAR BIOLOGY AND TRANSCRIPTOMICS

The Molecular Biology and Transcriptomics Unit carries out research in the field of Amyotrophic Lateral Sclerosis (ALS) and dementias (in particular Frontotemporal dementia -FTD) for genetic and transcriptomic characterization.

Genetic research is aimed at the study of complex phenotypes such as ALS-FTD, phenotypic overlapping conditions characteristic of neurodegenerative diseases, and the definition of the genetic profile as starting point of functional studies (by techniques such as Exomes sequencing and Sanger sequencing).

Transcriptomic research concerns the study of the role of RNAs as main regulators of numerous biological pathways implicated in the development and functioning of the brain and central nervous system. By transcriptomic techniques (RNA-Seq, Real Time PCR) it is possible to characterize the gene expression profile of the different classes of RNA, such as messenger RNA (mRNA) and non-coding RNAs (miRNA and lncRNA) in peripheral tissues, in cell lines and autopsy samples.

Related to transcriptomic, different studies of extracellular vesicles (EVs) in plasma from ALS and dementia subjects are in progress. EVs from plasma are characterized as diagnosis and prognosis markers in patients with ALS and dementia by the evaluation of RNAs and proteins cargo, focusing on the role of miRNAs in plasma EVs. Standard (immunofluorescence, real-time PCR and western blot) and innovative techniques (RNA-seq, confocal microscopy and Nanoparticle Tracking Analysis) are used for EVs characterization.

In recent years, the unit has also been dedicated to the study of R-loops in the pathogenesis of ALS. R-loops are three-stranded structures of DNA and RNA, which are formed during gene replication and transcription. The accumulation of R-loops can be a source of genomic instability and it has been shown that the motor neurons of ALS patients are unable to effectively repair this damage, leading to neurodegeneration. The unit is dedicated to the study of the molecular mechanism of R-loops linked to mutations of the TARDBP gene and to C9orf72 expansions in ALS patients and models.

Finally, the transcriptomics and genomic instability approach is also applied to the study of the Aicardi-Goutières syndrome (AGS), a rare genetic disease of neurodevelopment. Transcriptomic and phenotypic correlation studies are in progress for the definition of prognostic biomarkers and molecular mechanisms for the accumulation of R-loops in AGS patients.

The research activities are financed by various funding, such as the Ministero della Salute (Ricerca Finalizzata and Euronanomed), the Cariplo Foundation and ARISLA.