OVERVIEW

The course will provide an overview of the most advanced cell and gene therapies, starting from their molecular bases to their current translational applications. The course will deal with the fundamental biological concepts of stemness, pluripotency, plasticity and inducibility of the cells used in cell therapy approaches, and with the morphological and genetic characteristics of several viral families that makes them suitable as effective vectors for gene therapy. An introduction to genome-editing recent techniques will also be provided.

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 in the treatment of several pathological conditions, bringing students closer to a current research field and to its latest outcomes. We want also to evidence the deep changes brought by innovative advanced therapies to the concepts of "drug" and of "therapy" within the last two decades, including their positive and negative consequences. Moreover we also intend to stimulate the students' criticism not only relating to the technical contents of the subjects, but also focusing on the legislative mandatory requirements that define the use of cell and gene products in applicative medicine.

AIMS AND LEARNING OUTCOMES

At the end of the course students must retain: a) the general applicative principles, methodologies and technical requirements for the preparation of cell and gene therapy products; b) the biological, morphological, genetic and functional main requirements of the cell and viral types used in cell and gene therapy approaches. Students will have to be able to compare pros and cons and associated risks with advanced therapies, particularly in confrontation with current gold-standard therapeutic approaches. Students are also required to remember general rules and regulations defining production and use of cell and gene therapy products. We also expect that students will be able to evaluate comparatively different therapeutic approaches (i.e. traditional pharmacolgical vs advanced therapy) for the same pathological condition, critically supporting their observations and using the appropriate scientific terminology.

PREREQUISITES

For the best comprehension of the contents of the course, acquired competences in subjects such as molecular and cellular biology, biochemistry and physiology are strongly recommended –although not mandatory.

TEACHING METHODS

The course lasts 16 hours of front lessons, 8 of which are dedicated to cell therapy and the remaining eight to gene therapy. Lesson items are presented through PowerPoint presentations; didactic material (also non-textual) will be available under AulaWeb, through a controlled access system. During the course, students –if willing- may give a powerpoint presentation on a scientific paper -provided by the teacher and related to the course contents- with the aim to objectively summarize the clinical application described in the paper and critically evaluate its contents and conclusions. The presentation is non-mandatory, but it may be taken into consideration for the final evaluation of the student.

SYLLABUS/CONTENT

DETAILED PROGRAMS

CELL THERAPY (8 hours): Acquisition, characteristics, pros and cons and state-of-the-art in the research field for embrional and foetal stem cells, for stem cells derived from umbilical cord, placenta or amniotic liquid, and for adult stem cells, as well as induced pluripotent stem cells (iPSCs). Products for advanced therapies: definitions, general regulatory issues on their production, distribution and use. Cell therapy for the hemopoietic tissue: functional aspects of blood, of its cellular components and of the hematopoiesis. Collection, processing and cryopreservation of hemopoietic stem cells. Transplantation of hemopoietic stem cells: autologous or allogeneic translplantation. Cell therapy applications to ecto-mesodermal tissues: epithelial stem cells; cell therapy of the vascular tissues; cell therapy applications in orthopedics: cartilage and bone.

GENE THERAPY (8 hours): 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. Viral vectors used in gene therapy approaches: structure, characteristics, pros and cons; Herpesviridae (HSV):-Structural characteristics of HSV and of their life-cycle; immediate-early (IE), early (E) and late (L) genes and corresponding proteins; packaging systems based on helper virus and helper virus-free systems. Adenoviridae (AV):-Viral particle structure and its life cycle; 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). 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. Packaging cell lines bi- and tri-sected. Self-inactivating vectors; RV/LV use in gene therapy approaches.

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. General principles on engineered proteins to perform genome editing: Zinc finger Nucleases (ZFNs), Transcriptor activator-Like Effector nuclease (TALENs), and the CRISPR/Cas9 system.

RECOMMENDED READING/BIBLIOGRAPHY

Suggested textbooks:

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

BRUNO TASSO (President)

ELEONORA RUSSO (President Substitute)

LESSONS

LESSONS START

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

Class schedule

L'orario di tutti gli insegnamenti è consultabile all'indirizzo EasyAcademy.

EXAMS

EXAM DESCRIPTION

Before the formal final exam the student is invited to give a PowerPoint presentation during which he/she will expose the scientific results of a scientific publication dealing with the subjects of the course, in particular dealing with therapeutic approaches based on the technologies described during lessons. A critical overview, both relating methodology and results, is required; the presentation and its issues will be discussed publically among the students, his/her collegues and the teacher.

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

During the final oral examination, questions of raising difficulty will be posed, relating both major issues of the course (cell and gene therapy). Questions will assess knowledge of the functional and molecular components of cell and viral vectors used in therapy, as well as their application in clinical settings. 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 and the student's ability to correlate different topics; d) the terminology and language appropriateness; e) the student's criticism; in this respect the commission may also take into consideration the performance provided during the presentation of a scientific paper.

FURTHER INFORMATION

Additional info, relating the access keys to the material available within the AulaWeb platform as well as the scientific articles for the students' critical presentations, will be provided directly to the students during lessons.