E0230 Applied and Mechanistic in vitro Toxicology

Faculty of Science
autumn 2021
Extent and Intensity
2/1/0. 2 credit(s) (plus extra credits for completion). Type of Completion: zk (examination).
Teacher(s)
doc. RNDr. Pavel Babica, Ph.D. (lecturer)
RNDr. Iva Sovadinová, Ph.D. (lecturer)
prof. RNDr. Luděk Bláha, Ph.D. (lecturer)
doc. Mgr. Klára Hilscherová, Ph.D. (lecturer)
Guaranteed by
doc. RNDr. Pavel Babica, Ph.D.
RECETOX – Faculty of Science
Contact Person: doc. RNDr. Pavel Babica, Ph.D.
Supplier department: RECETOX – Faculty of Science
Timetable
Wed 15:00–16:50 D29/347-RCX2
Prerequisites
The course is intended primarily for Master students of Ecotoxicology specialization (preferentially in the 1st year of Master studies). An advantage is the previous attendance of at least basic ecotoxicology courses (E4070 Basics in Toxicology, E1240 Experimental and applied toxicology and ecotoxicology, E3230 Human pathophysiology, E0240 Biomarkers and toxicity mechanisms).
Course Enrolment Limitations
The course is also offered to the students of the fields other than those the course is directly associated with.
fields of study / plans the course is directly associated with
Course objectives
The aim of the course is to introduce basic mechanisms of toxicity at the sub-organismal level, including their mechanistic links to the adverse outcomes manifested at the level of individual or population. Students will learn about basic methods and approaches for assessment of the key mechanisms of toxicity and evaluation of the key biological processes by in vitro models and assays. The goal will be to introduce validated as well as pre-validated in vitro test used for assessment of specific adverse outcomes. In addition to principles and methodology of the specific in vitro models and tests, students will learn about the current legislative framework relevant for the application of in vitro assays for toxicity testing, as well as perspective directions and concepts which are currently being developed in predictive toxicology.
Learning outcomes
After completing the course, the student will be able:

• Describe basic mechanisms of toxicity at the suborganismal level and their links to adverse health outcomes at the individual or population level
• Explain principles of laboratory methods and models suitable for assessment of perturbations of the key toxicity mechanisms and disruptions of the key biological events occurring at the molecular, cellular and tissue level
• Understand principles and applications of in vitro assays validated or pre-validated for the assessment of specific adverse outcomes
• Understand testing guidelines, regulations and directives relevant for applications of alternative in vitro assays for toxicity testing
• Integrate results from different in vitro assays for purposes of risk assessment
• Discuss applicability of alternative vs. standard toxicity testing for practical and routine applications
• Elaborate on limitations and advantages of in vitro vs. in vivo methods for toxicity testing
• Describe measures, concepts, and approaches aiming to overcome current limitations of the in vitro assays and to increase their predictive value
Syllabus
  • • 1) Mechanisms of toxicity at the different levels of biological organization - molecular, cellular, tissue, organ, organismal / systemic toxicity. Introduction of the Adverse Outcome Pathways (AOPs) concept as a schematic representation of a series of causal and biologically relevant events (Key Events, KE) linking interactions of chemicals with molecular targets (Molecular Initiating Event, MIE) to an adverse outcome manifested at the individual / population level (Adverse Outcome, AO).
  • • 2-3) Basic mechanisms of toxicity at the suborganismal level:
  • a) Interactions of chemicals with biomolecules:
  • Genotoxicity and mutagenesis (mechanisms of direct and indirect DNA damage), interactions with biological membranes, interactions with receptors (membrane receptors, intracellular/nuclear receptors), interactions with enzymes, disruption of redox balance
  • b) Effects at the cellular and tissue level:
  • Cytotoxicity and viability, cell cycle and proliferation, programmed cell death, epigenetic regulations, disruption of differentiation and other cell events (GJIC, migration, invasivity, adhesion, immunomodulations), neoplastic transformation, Disruption of tissue homeostasis, the role of cell differentiation and stem cells in toxicity and chronic diseases (Barker hypothesis, stem cell theory of cancer).
  • • 4) Cell, tissue and organ in vitro models: basic definitions and types of in vitro models (primary cells, cell lines, organotypic cultures, explantates), their sources. Basic principles and techniques of in vitro cell cultures (adherent, suspension cell cultures), stem and pluripotent cells, co-cultures, 3-dimensional cell cultures, organoids, organ on-a-chip, body-on-a-chip.
  • • 5) Toxicity and hazard assessment, use of in vitro and other alternative approaches to predict adverse outcomes manifested at the organism level. 3Rs principle, terminology, comparison of in vivo and alternative in vitro methods and principles. Directive 2010/63/EU, validation process, ECVAM, OECD/EU TGs.
  • • 6) Overview and principles of in vitro and alternative methods used in toxicity testing and risk assessment - description of specific OECD/EU validated and prevalidate 3R methods and measures, their principles, the key mechanisms in the context of AOPs: dermal absorption, skin corrosion and irritation, eye corrosion and irritation, phototoxicity, immunotoxicity and skin sensitisation, genotoxicity and carcinogenesis.
  • • 7) Endocrine regulations and disruption, reproduction and developmental toxicity - methods for assessing MIE and KE - in vitro, gene transcription, biomarkers - hormon synthesizing or metabolizing enzymes, proteins (VTG), metaboites, validated and alternative in vitro approaches.
  • • 8) Further examples of alternative in vitro models for evaluation of the key events relevant for other mechanisms/AOPs for organ and systemic toxicity (toxic narcosis, neurotoxicity, hepatotoxicity, nefrotoxicity inhalation toxicity).
  • • 9) Legislative framework regulating use of alternative methods for toxicity testing: REACH, Cosmetic Directive, EC Reg. No. 1272/2008 GHS/CLP, 98/8/EC Biocides. Application of alternative methods within concepts of Integrated Approaches to Testing and Assessment (IATA) and Integrated Testing Strategies (ITS).
  • • 10) Toxicity testing in a high-throughput (HT) setup. Omics technologies - toxicogenomics, cellomics (HCA/HCS), examples of premier HT projects in toxicity testing (Tox21, ToxCast) and their applied outputs.
  • • 11) In vitro-in vivo extrapolation. Toxicokinetics and toxikodynamics in vivo and in vitro, in vivo dosimetry (direct/reverse), PBPK/PBTK. Basics of in vitro dose metrics, QIVIVE.
Literature
  • KUBINCOVÁ, Petra, Jiří NOVÁK and Iva SOVADINOVÁ. Acute Systemic Toxicity: Alternative in Vivo and in Vitro Methods. CHEMICKÉ LISTY. Praha: Česká společnost chemická, 2016, vol. 110, No 2, p. 118-125. ISSN 0009-2770. info
  • FOWLER, Bruce A. Molecular biological markers for toxicology and risk assessment. Amsterdam: Elsevier, 2016, ix, 153. ISBN 9780128095898. info
  • Casarett & Doull's essentials of toxicology. Edited by Curtis D. Klaassen - John B. Watkins. Third edition. New York: McGraw-Hill Medical, 2015, xiii, 524. ISBN 9780071847087. info
  • Predictive toxicology : from vision to reality. Edited by Friedlieb Pfannkuch - Laura Suter-Dick. Weinheim: Wiley-VCH, 2015, xxv, 404. ISBN 9783527336081. info
  • Comprehensive toxicology. Edited by Charlene A. McQueen - James Lamb. 2nd ed. Amsterdam: Elsevier, 2010, xvii, 267. ISBN 9780080468716. info
  • BOELSTERLI, Urs A. Mechanistic toxicology : the molecular basis of how chemicals disrupt biological targets. 2nd ed. New York: Informa Healthcare, 2009, 399 s. ISBN 9780849372728. info
  • Molecular and biochemical toxicology. Edited by Robert C. Smart - Ernest Hodgson. 4th ed. Hoboken, N.J.: John Wiley & Sons, 2008, xxxv, 901. ISBN 9780470102114. info
  • Principles and methods of toxicology. Edited by A. Wallace Hayes. 5th ed. New York: Informa Healthcare, 2008, xxiii, 227. ISBN 9780849337789. info
Teaching methods
The course will be conducted in a form of platform lectures accompanied with Powerpoint presentations. Lectures are interactive, with a use of audiovisual techniques and materials. Selected methods and approaches will be introduced in practical demonstrations and simple PC exercises (database searches, calculations). Students will be involved in discussions of the presented topic, with the emphasis on the understanding of mechanisms and their consequences.
Assessment methods
Attendance of the lectures is not mandatory but strongly recommended for complete understanding of the subject. The course is completed by a written exam, with a possibility of an follow-up oral exam.
Language of instruction
Czech
Further Comments
Study Materials
The course is taught annually.
The course is also listed under the following terms Autumn 2018, Autumn 2019, Autumn 2020, Autumn 2023.
  • Enrolment Statistics (autumn 2021, recent)
  • Permalink: https://is.muni.cz/course/sci/autumn2021/E0230