OVERVIEW

The course will provide an overview of the most advanced gene therapies, starting from their molecular bases to their current translational applications. The course will deal with the fundamental biological and morphological characteristics, with the genetic content and organization and with the integrative competence of several viral families that makes them suitable as effective vectors for gene therapy.

AIMS AND CONTENT

LEARNING OUTCOMES

To allow the student to acquire a detailed knowledge of the current and up-to date gene transfer and therapy approaches, along with their evolution in clinical settings, and to correlate them with the current knowledge on physiology, molecular biology and genetic engineering technologies; to allow the student to envisage possible future applications of gene therapy based on the current knowledge of monogenic diseases with a relevant clinical prevalence.

LEARNING OUTCOMES (FURTHER INFO)

At the end of the course the student must retain: a) the general applicative principles, methodologies and technical requirements for the preparation of gene therapy products; b) the biological, morphological, genetic and functional main requirements of the viral types used gene therapy approaches; c) pros and cons and associated risks with gene therapies, particularly in confrontation with current gold-standard therapeutic approaches. We also expect that the student will be able to evaluate comparatively different therapeutic approaches (i.e. traditional pharmacological  vs advanced therapy) for the same pathological condition, critically supporting his/her observations.

TEACHING METHODS

The course lasts 20 hours of front lessons, during which lesson items are presented through PowerPoint presentations; the teacher will provide the students with didactic material. During the course the students will be offered a chance to give a powerpoint presentation on a scientific paper provided by the teacher. The presentation is non-mandatory, but it may be considered for the final evaluation of the student.

SYLLABUS/CONTENT

DETAILED PROGRAMS

GENE THERAPY:Clinical trials based on gene therapy: geographical distribution and socio-economic aspects. General principles of gene therapy and choice of the proper target pathology. Side effects: consequences and complications. Examples of therapeutic approaches by means of gene therapy (for ex. in hematopoietic cells, in muscular dystrophy, in tumor treatment or in neuropathological disorders) will be presented along the description of the viral vectors used (Adenosine deaminase deficiency, SCID and X-SCID, Junctional Epidermolysis bullosa).

Viral vectors used in gene therapy approaches: structure, characteristics, pros and cons

Herpesviridae (HSV):-Structural characteristics of HSV capsid, matrix and envelope; structural organization of the viral genome; immediate-early (IE), early (E) and late (L) genes and corresponding proteins; HSV lifecycle (lytic and lysogenic cycles); viral tropism; packaging systems based on helper virus and helper virus-free systems; transgene insertion in HSV; optimization for targeting: gB and gD functions. -Sindbin Virus e Semliki Forest Viruses (SFV) as suicidal vectors.

Adenoviridae (AV):-Viral particle structure; genome structure; E1, E2, E3 and E4 genes; ITR and psi (ψ) region of the viral genome; effects of AVs and specific functions of their proteins against host defenses. AV vectors and their potentiation (promoter-specific activities, pro-drug enzymes, vector-spread molecules, anti-tumor molecules). Pre-clinical aspects: positive and negative results on murine experiments. Adeno-associated viruses (AAV) and applications of AV/AAV approaches.

Retroviridae(RV)-Viral particle structure of retroviruses; the RV genome and its integrated provirus; main relevant characteristics of the RV genome: gene products, LTRs, cPPT, TAR, RRE. Replication of the viral genome; generation of helper viruses for packaging cell lines; minigene vectors; Packaging cell lines bi- and tri-sected; -Accessory proteins in Lentiviridae (Tet, Rev, Vif, Vpi, Vpx, Vpu and Nef); SMAR sequences. Self-inactivating vectors (SIN).

Current methods for the recovery transgene-carrying functional viral particles: uòtracentrifugation, poly-lysine precipitation, ionic-exchange chromatography. Comparison between HSV, AV, AAV, RV and LV. Use of LV for shRNA transfer.

Non viral-based gene therapy approaches and principles of “genome editing”:Naked DNA inoculation, jet-gas injection, gene-gun, electroporation, sonoporation, hydrodynamic transfer, chemical vectors. Characteristics of target tissues for non-viral based gene transfer.

RECOMMENDED READING/BIBLIOGRAPHY

F. Zinno; “Principi di terapia cellulare”; e-book; Amazon.it

M. Giacca; “Terapia Genica”; Springer Biomed Eds., 2011

A.J. Dunn; “Elementi di virologia molecolare”; CEA Eds.

B. Alberts,et al; “Biologia molecolare della cellula”; Zanichelli Eds.

J.D. Watson, et al.: “DNA Ricombinante”; Zanichelli Eds.

TEACHERS AND EXAM BOARD

Exam Board

PAOLO GIANNONI (President)

GRAZIA MARIA SPAGGIARI (President)

LESSONS

LESSONS START

Location, date and timetable of the lessons will be provided by the competent office.

Class schedule

The timetable for this course is available here: Portale EasyAcademy

EXAMS

EXAM DESCRIPTION

The student's performance is evaluated during an oral examination (average duration: 30 min), according to the exam calendar and timetable set by the competent office.

ASSESSMENT METHODS

Questions of raising difficulty will be posed, relating major issues of the course In particular we will evaluate: a) exactness of the student's response; b) the student's ability to frame the topic in a wider context; c) the student's ability to correlate the topic with other topics, not necessarilty related to the course; d) the terminology and language appropriatness; e) the student's criticism. For the final evaluation of the candidate the commission may also take into consideration the performance provided during the non-mandatory presentation of a scientific paper.

Exam schedule

FURTHER INFORMATION

The scientific articles for the students' critical presentations, will be provided directly to the students during lessions.