Neurogenetics

The Neurogenetics Programme was set up to ensure the enhancement and integration of neurogenetic and molecular neurobiology diagnostic activities within the diagnostic pathways for hereditary neurological diseases and the related research and training activities, with particular regard to technological innovations.

• The Neurogenetics Programme ensures the introduction and application of the most advanced methods of:
• Genetic analysis with next-generation sequencing (NGS), based on analysis of the exome and of panels of target genes, and with direct sequencing of disease genes and validation of the results obtained with NGS methods, both of the nuclear and mitochondrial genome;
• in vitro functional analyses using qualitative and quantitative biochemical methods on primary cells (fibroblasts) and/or tissues (muscle biopsies) derived from the patients under investigation, integrated with molecular biology methods applied to cellular models, including reprogramming to pluripotent stem cells and differentiation of cell and/or tissue types targeted by the disease.

The main techniques for the molecular diagnosis of hereditary neurological diseases

• Sanger sequencing
• NGS sequencing
• Fragment analysis (MLPA, Minisequencing, expansions)
• Absolute quantification by qPCR and ddPCR

The main techniques for functional validation and for studying the molecular mechanisms of neurogenetic diseases for diagnostic and therapeutic purposes

• Biochemical diagnostics on primary cells and/or tissues (muscle biopsy, lymphocytes) of mitochondrial respiratory chain enzymes and other enzymes of mitochondrial metabolism (thymidine phosphorylase and nucleotide assay)
• Study of primary patient cell lines (fibroblasts/myoblasts)
• Cell validation respirometry
• Reprogramming of stem cells from primary patient cells (iPSCs)
• Differentiation of cell types or disease target tissues
• Transcriptomics and gene expression profiling on patient-derived tissues and cellular models
  of disease, including single cell analysis
• Design, testing and validation of therapeutic, pharmacological and/or advanced therapy strategies (gene therapy)

Diagnostic and laboratory research activities are carried out in close relation to the clinical and genetic counselling activities carried out at the Institute thanks to the participation in the PDTAs and protocols/pathways of care active at the ISNB, providing an innovative and technologically advanced contribution to the achievement of the objectives of all four research lines of the Institute.

The Programme is the regional reference for molecular diagnostics of mitochondrial diseases and the ISNB's reference for the Genomics platform of the IRCCS Neuroscience and Rehabilitation Network (RIN) as well as for the genomics activities of the National Virtual Institutes for Dementia and Parkinson's Diseases and the new institutes to be set up.

The Programme contributes to university training activities at I, II, III level and to refresher courses for NHS professionals.

The main neurogenic diseases to which the programme's activities apply are:

• Mitochondrial diseases: mitochondrial encephalomyopathies from defects in mitochondrial DNA (MELAS/MERRF/NARP/KSS); hereditary optic atrophies (LHON/DOA/ROA/X-linked);
  encephalomyopathies and rare neurodegenerative syndromes from nuclear DNA defects involving genes encoding mitochondrial proteins
• Rare neurological diseases: this includes rare metabolic diseases, both paediatric and adult, including spastic paraparesis and erythroderma.
• Neurodegenerative diseases: this includes Parkinson's disease and Parkinsonisms, Amyotrophic Lateral Sclerosis.
• Epilepsy syndromes: genetic epilepsies in childhood and adults.