CODE 98815 ACADEMIC YEAR 2018/2019 CREDITS 2.5 cfu anno 2 MEDICAL-PHARMACEUTICAL BIOTECHNOLOGY 10598 (LM-9) - GENOVA SCIENTIFIC DISCIPLINARY SECTOR BIO/13 LANGUAGE English TEACHING LOCATION GENOVA SEMESTER 2° Semester TEACHING MATERIALS AULAWEB 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 The aim of the course is to evidence the applicative potential displayed by some technological innovations in the fields of cellular and molecular biology and in the knowledge of the vital cycle of several viral families, bringing students closer to a current research field and to its latest outcomes. We want also to evidence the deep changes brought by innovative gene therapies to the concepts of "drug" and of "therapy" within the last two decades, including their positive and negative consequences. AIMS AND LEARNING OUTCOMES 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 24 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 Suggested textbooks: 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 PAOLO GIANNONI Ricevimento: Office hours for students are normally set on Thursdays from 09:00 to 11:00. The office address is: Biology Section, Dept. Experimental Medicine, entrance: Via Pastore 3 or C.so Europa 30, second floor. E-mail appointments are strongly encouraged. The teacher's e mail address is the following: paolo.giannoni@unige.it. Alternatively the teacher can be reached at the following phone number: 01035338201. Exam Board PAOLO GIANNONI (President) RODOLFO QUARTO LESSONS LESSONS START The course is planned for the second semester. 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. FURTHER INFORMATION The scientific articles for the students' critical presentations will be provided directly to the students during lessons.
