CONOSCENZE E ABILITÀ DA ACQUISIRE

The first part of the course aims to provide students with notions on gene therapy applied to the resolution of human hereditary and non-hereditary pathological phenotypes. The course is mainly devoted to the presentation of problems of medical genetics that can be potentially faced by biotechnological approaches.

The second part of the course addresses current aspects of stem cell biology and epigenetic gene regulation. Epigenetic modification and regulation of stem cells in normal development and in pathology will be discussed.

Students will have knowledge on basic principles and applications of the most important molecular methodologies with a particular emphasis to genetic and epigenetic methods.

PROGRAMMA/CONTENUTI DELL’INSEGNAMENTO

MODULO: Diagnostics and animal model of genetic diseases

Major technological approaches for analysis of nucleic acids are presented as well as general techniques for generation of animal models of genetic diseases.

Specific issues:

1. General features of diseases with  monofactorial and multifactorial genetic components.

2. Relevance of SNPs and CNVs evaluation in medical genetics

3. Technological approaches to evaluate SNPs and CNVs

4. Morphological and molecular caryotyping

5. DNA sequencing

6. Complex diseases and the problem of missing heritability

7. Epigenetics in human diseases

8. Animal models of genetic diseases2.      

MODULO: Approaches for Gene therapy, Next Generation Sequencing  and  multifactorial diseases (Prof. Aggr. Lonigro)

PART I: Gene Therapy, (Prof. Aggr. Renata Lonigro)

Gene therapy applied to the resolution of human hereditary and non-hereditary.

pathological phenotypes.

Methods of gene therapy, advantages and disadvantages :

- Gene therapy ex vivo and in vivo

- Additives and ablative strategies

Transduction methods based on the use of viral vectors:

- Production of viral vectors and cells packaging

- Principal types of viral vectors (retroviral, adenoviral, adeno-associated, and lentiviral derived SIN vectors)

- Systemic administration of therapeutic genes and cell targeting

- Monitoring the temporal and spatial expression of the transgene

- Gene therapy based on non-viral transduction methods

Preclinical and clinical studies: animal models of disease and some clinical trials performed in humans.

- Gene therapy of cancer

- Gene therapy directed to the myocardium

- Gene therapy of neuromuscular diseases such as DMD / BMD, LGMD and SMA

PART II: gene sequencing and NGS methods (Prof. Aggr. Renata Lonigro)

- General description of methods and applications of Next Generation Sequencing (NGS)

- Strategies for the construction of gene-specific libraries and an Illumina-based platform for NGS   sequencing: the pilot project on Spinal Muscular Atrophy.

- Identification of SNPs and allelic variants: future implications for diagnosis / prognosis of a mendelian disease and for the planning of pharmacogenetic studies.

PART III: Genetics of multifactorial diseases (Prof. Aggr. Renata Lonigro)

Genetics of complex traits:

- Heritability and environment in human diseases

- Defining the susceptibility to the onset of a multifactorial disease

- Predisposing genes, their interaction and inheritance

- Studies on twins and concordance index (IBD and IBS)

- Analysis SPA and TDT

Strategies to identify genes predisposing to the onset of a multifactorial disease:

- Parametric and non-parametric studies

- Genome Wide Association Studies (GWAS)

- Animal models of complex diseases (i.e. atherosclerosis)

Gene mutations identified as predisposing to the onset of some multifactorial diseases (i.e.breast cancer, diabetes, hypertension, atherosclerosis, schizophrenia and bipolar disorder)

PARTE IV: Introduction to NGS sequencing – focus on the Illumina platform

Theoretical and practical lessons headed by other teachers

Lesson 1 - 3 hrs      dr. Giusi Zaina

Introduction to NGS sequencing – focus on the Illumina platform [2 h]

Sample preparation – NGS library [1 hr]

Lesson 2 - 3 hrs dr. Simone Scalabrin

Overview of the Illumina pipeline and Raw sequence file format [1 h]

Get summary statistics before alignment [1 h]

Data cleaning: trimming and filtering [1 h]

Lesson 3 - 3 hrs dr. Slobodanka Radovic

Targeted re-sequencing for research and diagnostics [1 h]

Identification of disease-associated variants: trio analysis [2 h]

MODULO: Epigenetics and Stem cells

The course addresses current aspects of stem cell biology and epigenetic gene regulation.

The program covers:

• key mechanisms that control pluri- and multipotency of stem cells; differences between Embryonic and Adult stem cells.
• Mechanisms and models of somatic cell reprogramming (iPS).
• Central epigenetic regulatory mechanisms that include transposons, pseudogenes, imprinting, gene silencing, X-chromosome inactivation, position effect, reprogramming, regulation of DNA and histone modifications, heterochromatin and technical limitation affecting reproductive cloning; epigenetics and diseases.
• Molecular biotechnologies to study epigenetics
• Current advancements in understanding the role of the non-coding genome, focusing on long and small non coding RNAs and pseudogenes.
• Particular focus is given on stem cell-based regenerative medicine and its great promise for repair of diseased tissue.
• The origin of Cancer: Stemness and Tumorigenesis.

Teaching materials: ppt. available and literature search

ATTIVITÀ DI APPRENDIMENTO E METODI DIDATTICI PREVISTI

The program includes lectures, seminars and pratical exercises.

MODALITÀ DI VERIFICA DELL’APPRENDIMENTO

Written exam on knowledge and understanding of the topics addressed during the course. 

TESTI/BIBLIOGRAFIA

 PDFs and other literature material.

STRUMENTI A SUPPORTO DELLA DIDATTICA

PDFs and other material will be provided to the students.

TESI DI LAUREA

Master thesis projects that are in line with the addressed topics will be available in the labs of Proff. Damante, Lonigro and Benetti.