Genetics and moleculars bases of diseases

A.Y. 2015/2016
Overall hours
BIO/13 MED/03
Learning objectives
Relevant objectives of the course are the acquisition by the students of competences concerning the organization of the human genome and the genetic and epigenetics mechanisms involved in genome mutation and in control of gene expression. The molecular mechanisms learned in the first part of the course will help to understand the regulation of specific function of the immune system and the pathogenesis of the immune diseases with perspective of gene therapy. These issues will allow to achieve other relevant goals of the course: the comprehension of molecular mechanisms underlying both constitutional monogenic and poligenic diseases and cancer. Furthermore, experimental activities in labs, concerning modern methods applied in molecular diagnosis of genetic diseases, will be carried out by each student.
Expected learning outcomes
Course syllabus and organization

Single session

Lesson period
Second trimester
Modulo: Biologia applicata
Course syllabus
Organization of the human genome
1. Human multigene families
2. Extragenic conserved sequences
3. Extragenic and coding repeated DNA sequences
4. Transposable elements
Struttura del nucleo interfasico
Instabilità del genoma
1. Mutations and polymorphisms of DNA
2. Genetic mechanisms underlying mutation of repeated sequences
3. DNA repair
4. Segmental duplications, CNVs and the causative molecular mechanisms
Mutazioni patogenetiche
1. Classification and database nomenclature of mutations and databases of mutations
2. Loss and gain of function mutations
3. Pathogenetic mutations and the pathogenic potential of repeated sequences

Molecular mechanism of epigenetics
1. DNA methylation
2. Histone modifications and histone variants
3. ATP-dependent chromatin modifications
4. Tridimensional nuclear organization
Epigenetic thecnologies
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 cellular differentiation
Epigenetics and human pathologies
Epigenetic therapies

Memory in the innate and adaptive immune system
1. Epigenetics of the innate immune response: cell differentiation, stimulation and memory. Selectivity of the transcriptional response to stimulus.
2. Contribution of chromatin to cell memory.
3. Cells of the adaptive immune system.
4. Epigenetics of cell identity in the adaptive immune system.
Role of AIRE in the physiology and pathophysiology of the immune system
1. The thymus as lymphopoietic organ.
2. T cell physiology. "Autoimmune regulator" (AIRE) in the regulation of ectopic tissue antigens expression in the thymus. The APECED syndrome ("Autoimmune polyendocrinopathy candidiasis ectodermal dystrophy") by mutations of AIRE.
Primary immunodeficiencies
1. Classification of primary immunodeficiencies.
2. Immunodeficiencies with dysregulation of the immune response.
3. Immunodeficiencies with impaired cell-mediated cytotoxicity.
4. Defects in the innate immune system.
Induced pluripotent stem cells as cellular model for the study of diseases
1. Factors inducing stem cell pluripotency.
2. Examples of disease studied using induced pluripotent stem cells.
Gene therapy
1. Principles of gene therapy.
2. Vectors for gene therapy. Retroviral, lentiviral, adenoviral vectors; vectors based on adeno-associated and herpes simplex virus.
3. Disorders approached by gene therapy.
4. Possible complications of gene therapy.
Possible future of gene therapy
Modulo: Genetica medica
Course syllabus

Framing and load of genetic diseases
Molecular genetics of neuromuscolar diseases
1. Duchenne e Becker muscular dystophy
2. Amyotrophic lateral sclerosis (SLA)
Molecular genetics of neurological diseases
1. Charcot-Marie-Tooth (CMT) disease
2. X-linked mental retardation(XLMR)
3. Members of the MARK family kinases: between Alzheimer and
Dynamic mutation syndromes
1. X fragile (FRAXA) syndrome
2. Huntington disease(HD)
3. Myotonic dystrophy (DM1, DM2)
4. Friedreich ataxia (FD)
5. Spinocerebellar ataxia
Genomic imprinting and related syndromes
1. Beckwith-Wiedemann syndrome
2. Angelman e Prader-Willi syndromes
Molecular genetics of mitochondrial diseases
1. Leber neuropathy(LHON)
2. Neuropathy, ataxia and retinitis pigmentosa (NARP)
Molecular genetics of cancer syndromes
1. "Cancer genes"
2. Cancer predisposition syndromes: HNPCC, FAP, RTS, BS, WRN, and Li-
3. Genomic instability and tumorigenesis
4. MVA syndrome
5. Trisomy 21 and predisposition to megakaryoblastic leukemia
6. A-to-I RNA editing: the "ADAR" side of human cancer
Molecular genetics of hematological malignancies
1. Molecular events in Acute Myeloid Leukemia
2. Genetic and molecular heterogeneity in T-cell non-Hodgkin lymphoma
3. miRNA roles in leuchemogenesis
In each lesson one hour will be dedicated to:
Biotechnological applications in Medical and Molecular Genetics
1. Approaches for disease-causing sequence variants identification
2. Specificity: a very important factor for detection of rare events
3. Principles and paradigms in expression analysis
4. Quantification: about sensitivity, dynamic range and precision
5. Stem Loop RT-PCR in miRNA expression quantification
6. qRT-PCR
7. Microarrays: methodologies and analysis approaches
8. Droplet digital PCR (ddPCR)
9. Sequencing: from Sanger to "next generation"
10. Molecular genetics at a single molecule resolution
11. Circular probes and RCA (rolling circle amplification) principles
12. mRNA detection and genotyping in situ
13. in situ mRNA sequencing
14. Protein Ligation Assay: principles and evolution
Modulo: Biologia applicata
BIO/13 - EXPERIMENTAL BIOLOGY - University credits: 8
: 48 hours
Informal teaching: 30 hours
Modulo: Genetica medica
MED/03 - MEDICAL GENETICS - University credits: 6
: 48 hours
Professor: Beghini Alessandro
Appointment upon request
Dept. Medical Biotechnology and Translational Medicine, Via F.lli Cervi 93, 20090 Segrate