Genetic and molecular bases of diseases

ORGANIZATION OF THE HUMAN GENOME
1. Human multigene families
2. Extragenic conserved sequences
3. Extragenic and coding repeated DNA sequences
4. The coding genome

GENOME INSTABILITY
1. Mutations and polymorphisms of DNA
2. Genetic mechanisms underlying mutation of repeated sequences
3. Segmental duplications, CNVs and the causative molecular mechanisms

MOLECULAR PATHOGENETIC MECHANISMS
1. Classification and database nomenclature of mutations
2. Loss and gain of function mutations
3. Pathogenic potential of repeated sequences
4. The chromothripsis
5. The variable expressivity and the expression variability: the Noonan syndrome as model disease

MOLECULAR MECHANISM OF EPIGENETICS
1. An introduction to epigenetics
2. Role of Epigenetics in development and cell differentiation
3. Examples of different epigenetically-regulated phenomenon
4. Molecular mechanism of epigenetics: Covalent Histone modifications, ATP-dependent chromatin remodeling, histone variants.

EPIGENETIC TECHNOLOGIES
1. Functional characterization of regulatory DNA elements (promoters, enhancers, silencers, insulators)
2. Methods to study DNA-protein interactions in vitro.
3. Chromatin immunoprecipitation and in vivo applications.

EPIGENETICS AND HUMAN PATHOLOGIES
1. Epigenetics and neurodegenerative disease and neuropsychiatric disorders
2. Epigenetics and memory
3. Epigenetics and cancer

EPIGENETIC THERAPY

Medical Genetics
GENOMIC DISORDERS
1. Recurrent and non-recurrent genomic rearrangements and disease traits
2. Copy Number variation: from inter-individual genetic variability to disease
3. Array CGH for the analysis of constitutional genomic rearrangements. How interpret the findings

CNVS SIGNIFICANCE ASSESSMENT FLOW CHART: CASE STUDIES
Learning activities through acquisition, inquiry, collaboration, discussion and production

GENOMIC IMPRINTING AND RELATED SYNDROMES
1. The 15q11.2-13 imprinted region and related syndromes (Prader-Willi and Angelman syndromes)
2. 11p15.5 related syndromes (Beckwith-Wiedemann, Silver Russell syndrome, IMAGE)
3. Multilocus Imprinting Disorders

MOSAICISM IN HUMAN DISEASES: FROM CNVS TO SNVS

PRINCIPLES OF MOLECULAR DISEASES
1. Effect of mutations on protein function
2. Examples of Mendelian disorders illustrating mechanism by which mutations produce disease in different classes of proteins

Exploring Single Nucleotide Variants (SNVs) effect
Learning activities through acquisition, inquiry, collaboration, discussion and production

PENETRANCE DEFECTS AND PHENOTYPE VARIABILITY: TOWARDS AN UNDERSTANDING OF THE MOLECULAR BASIS
1. TAR syndrome and the two hits condition
2. Velo Cardial Facial syndrome
3. Marfan syndrome

FROM MONOGENIC TO "COMMON" DISEASES
Type 2 Diabetes

THE POSITION EFFECT: WHEN THE MUTATION DOES NOT AFFECT CODING GENE SEQUENCES
MITOCHONDRIAL MEDICINE
1. Mitochondria and mitochondrial (dys)functions
2. Mitochondrial disorders caused by defects of nuclear DNA
3. Mitochondrial disorders caused by defects of mtDNA
4. Mitochondrial dysfunctions in neurodegenerative diseases
5. Case studies: patients with mtDNA mutations (point mutations or mtDNA deletion/depletion) vs patients with nuclear mutations

CANCER GENETICS
1. Sporadic and inherited cancer predisposition
2. Ovarian and breast cancer predisposition
3. Li-Fraumeni syndrome, Cowden syndrome
4. Colon cancer predisposition syndromes
5. MEN syndromes
6. Renal cancer predisposition

NEUROMUSCULAR DISORDERS
1. Muscular dystrophies
2. Spinal Muscular Atrophy
3. Amyotrophic Lateral Sclerosis
4. New therapeutic approaches for neuromuscular disorders

CASE DISCUSSION

MOLECULAR THERAPIES
1. Main principles of gene therapy
2. Modulation of gene expression
3. Induction of Exon Skipping/Retention - Splicing modulation (ASO)
4. Gene Editing (CRISPR/Cas9)
5. Gene therapy vs. gene editing

Practical laboratory activities
'Methods for the identification of DNA sequence variations'
LECTURES
1. Overview of the main techniques for the detection of point mutations, small deletions and small insertions
2. Direct Sanger sequencing and microsatellite analysis by capillary electrophoresis
3. Methylation-specific PCR

PRACTICAL LABORATORY ACTIVITIES
1. PCR amplification of genomic DNA fragments
2. Analysis of PCR fragments by gel electrophoresis
3. Direct Sanger sequencing
4. Microsatellite analysis
5. Methylation-specific PCR
6. Real-Time PCR