Unit NON-VIRAL VEHICLES FOR GENE THERAPY
- Course
- Pharmaceutical biotechnologies
- Study-unit Code
- GP003551
- Location
- PERUGIA
- Curriculum
- In all curricula
- Teacher
- Stefano Giovagnoli
- Teachers
-
- Stefano Giovagnoli
- Hours
- 42 ore - Stefano Giovagnoli
- CFU
- 6
- Course Regulation
- Coorte 2022
- Offered
- 2022/23
- Learning activities
- Caratterizzante
- Area
- Discipline farmaceutiche
- Academic discipline
- CHIM/09
- Type of study-unit
- Obbligatorio (Required)
- Type of learning activities
- Attività formativa monodisciplinare
- Language of instruction
- Italian
- Contents
- Introduction pharmacokinetic concepts Site-specific drug delivery. Concepts of gene therapy. Hints about viruses like vectors. Naked DNA bundling or nanoparticulate systems. Complex DNA lipid, DNA polymer. Composite nanoparticles and their preparation
- Reference texts
- The material needed is provided by the teacher- slides and scientific publications in international journals
- Educational objectives
- The course provides a theoretical knowledge of molecular systems and nanoparticles used as carriers in gene therapy alternative to viral.
The knowledge-knowledge that will be acquired by the student concerns the technological basis of gene therapy. in particular, students will have to demonstrate that they understand the nature of colloidal systems in particular nanoparticles and molecular carriers used as vectors of genetic material, the problems related to their preparation and stability, the differences between the different lipid and polymer systems, the different mechanisms of action of the carriers described, and the characteristics of use required for ensuring maximum effectiveness of the treatment.
Skills- students must obtain the decision-making capacity on the choice of appropriate carriers depending on the type of target and the type of genetic disease. In addition, they will have to develop the ability to learn on their own the topic and discuss the literature of the field. - Prerequisites
- In order to ensure a sufficient level of learning, the student must have acquired knowledge of organic chemistry and inorganic chemistry. In addition, basic knowledge of biochemistry and physiology and histology are fundamental. In particular, the course requires a basic understanding of cell biology and cellular metabolic processes as well as general human anatomy. This knowledge is important for the multidisciplinary nature of the course ranging from biological systems to the technologies applied to pharmaceutical forms. In this regard, a study of the basic concepts and approaches useful to drug delivery is suggested. Some of these aspects will be anyway addressed at the beginning of the course.
- Teaching methods
- The course consists of classroom lectures on topics with special attention to involve the student to retrieve and evaluate the basic knowledge necessary for learning. For this purpose, the slides of the course and literature on recent scientific advances in the field will be provided. This is believed necessary given the lack of a reference text to didactic scope of the student. Both the multi-disciplinary course and the advanced level of the didactic material that the student has available require continuous support from the teacher to verify that the student has the right level of understanding of the topics discussed. In addition, the course includes a period of continuous updating due to the rapid progress in the field that is carried out through analysis of the scientific literature. To this end, a second step involves a study that will be done in the hours dedicated to support the course.
- Other information
- Support activities will be carried out after the course to develop certain aspects of the course to be agreed with the students and help the students in the activity of connection of prior knowledge and necessary learning and new knowledge gained during the course.
This activity will be held at the Center of Biotechnology in via del Giochetto - Learning verification modality
- Oral exams, consisting of questions relating to the theoretical aspects concerning the topics covered during the course and aimed at demonstrating the knowledge and understanding of the student of such aspects, and the ability to expose the course contents; in particular, the student will have to demonstrate that they understand the nature of colloidal systems in particular nanoparticles and molecular carrier used as vectors of genetic material, the problems related to their preparation and stability, the differences between the different lipid and polymer systems, the different mechanisms of action of the carriers described, and the characteristics of use required for ensuring maximum effectiveness of the treatment. In addition,the ability of the student to learn on their own the topic and discuss the literature of the field will be examined.
- Extended program
- Introduction: pharmacokinetic concepts and in particular biodistribution-site-specific delivery of drugs and active molecules: definitions and approaches- The principle of vectorization and concepts and technology of colloidal dispersed systems. Properties of nanoparticles and pharmacokinetics. Hints on viruses as vectors: properties and typology of viral vectors and preparation methods. DNA delivery via liposomes. Types of liposomes and their properties and methods of preparation. Advantages and disadvantages. pH-sensitive liposomes (activosomes).
Lipid complexes and DNA: Lipoplexes (genosomes): types of lipids and properties. Cationic lipids and mixtures, helper lipid. Preparation methods. Classical lipoplexes and endosomal escape mechanisms (flip-flop). Structure of lipoplexes. Plasmid DNA delivery: SPLP. LDP lipid particles. Use of polycation and condensation: LPDI and LPDII particles.
Polyplexes: polymers and properties, condensation and polyplexes typology. Endosomal Escape: proton-sponge process and properties of polymers. PAM, PAMAM, PEI. Copolymers. Branching and effect on transfection. Advantages and disadvantages of polycations. Dendrimers and their properties. Methods to improve biocompatibility. Quaternization and hydroxylation.
Delivery of naked DNA. Advantages and advantages, sequential administration.
Cell penetrating peptides. Nature and properties, type and operation. Mechanisms of entry into the cell. Moment of hydrophobicity. Fusogenic peptides. Multivalent systems and examples of control on the endosomal escape process. Advantages and problems.
Nuclear DNA transduction: NLS, DTS and transport mechanism in the nucleus.
Organic and inorganic and mixed polymeric nanoparticles. Examples, advantages and disadvantages. PLGA and use of mesoporous silicates. Carbon nanotubes and alloysites. Metal nanoparticles: iron (magnetofection).
Gold nanoparticles: characteristics, preparation methods and properties. Bond with DNA and types of NP. Surface plasmon resonance and thiolation. Advantages and disadvantages. Quantum-dots.
Other applications: Gene-gun.