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| Preclinical Anticancer Drugs Screening (PADS) Based on Novel Models of Cancer BiologyJuan Sebastian Yakisich, MD, PhD | |||||||||||||
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Syllabus |
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Instructor: Juan Sebastian Yakisich |
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Content Outline for Preclinical Anticancer Drugs Screening (PADS) Based on Novel Models of Cancer BiologyPADS is an interdisciplinary course that integrates current concepts of cell biology and pharmacology with basic, translational and clinical oncology with the aim to provide students a comprehensive theoretical knowledge and laboratory experience in the field of preclinical anticancer research. |
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e-mail: yakisich@gmail.com |
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GOALS |
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OVERVIEW: The content of the lectures and laboratory practice are specially selected to introduce leading concepts in the field and cutting edge technologies. The course is organized as a hypothesis-driven research project that will evaluate the anticancer properties of selected compounds. Fourteen lectures delivered by leading researchers in their field are integrated with 6 weeks (two days/week) of extensive laboratory practice. The students will be involved in the design, execution, analysis and interpretation of experiments resembling real life scientific research. |
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TEXT & REFERENCE MATERIALS Selected scientific articles will be provided |
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Prerequisite: Cell Biology, Molecular Biology, Chemistry, |
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SPECIFIC COVERAGE
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Lecture 1 Overview of the course and the anticancer drugs screening process Overview of the course. Preclinical studies. History. Ethical considerations. Clinical Studies. Phases 0-IV. History: Ethical considerations.
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Lecture 2 Cell Culture Cell-based methods to screen compounds with anticancer activity Cell lines. Commercial and patient derived cell lines. Two and three dimensional systems. Cancer stem cell systems. Short-term assays (e.g. MTT). Advantages and limitations. Long term assays (e.g. Colony forming assay). Advantages and limitations. High throughput assays.
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Lecture 3 Cell Cycle I Overview of the cell cycle in normal and cancer cells Phases of the cell cycle. Cell cycle checkpoints. Dysregulation of the cell cycle in cancer cells. Cell cycle regulators as target for anticancer drugs.
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Lecture 4 Cell Cycle II Methods to study the mammalian cell cycle Methods for cell cycle synchronization (e.g. serum withdrawal, pharmacological arrest, elutriation) Method to assess cell cycle duration and arrest (FACS, 3H-thymidine labeling)
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Lecture 5 Cancer Biology I Basic Aspects of Cancer Cells Characteristics of cancer cells. Carcinogenesis. Metastasis. Mechanisms of drug-resistance.
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Lecture 6 Cancer Biology II Cancer Stem Cell Theory and Alternative Models Stem cell theory. Methods to isolate and charactherize putative cancer stem cells. Tumor microenvironment Alternative models of cancer stem cells. Implications for clinical trials and anticancer drus screening strategies.
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Lecture 7 Drug Effects I Methods to study drug effects Dose-response curves. Determination of the IC50 and RC0 and other endpoints Drug interactions (e.g. synergism).
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Lecture 8 Drug Effects II System assays and endpoint parameters Continuation of Lecture 2: cell-based assays. Concentration-response curves. Survival plots. Endpoint parameters. Uses. Advantages and limitations (e.g. IC50, RC0, OS, PFS)
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Lecture 9 Programmed Cell Death I Programmed Cell Death in Normal and Cancer Cell Overview of programmed cell death (PCD). Apoptosis. Autophagy. Necrosis. Autoschizis. Ferroptosis. Signalling pathways. Crosstalk between different types of PCD. Novel cell death subroutines: "anoikis," "paraptosis," "pyroptosis," "pyronecrosis" Mitotic catastrophe PCD as target for anticancer drug
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Lecture 10 Programmed Cell Death II Methods to Study Apoptosis Methods to study apoptosis. Principles. (e.g. DNA laddering, Hoechst staining, PARP, Annexin V, caspases). Guidelines for the use and interpretation of assays for monitoring apoptosis
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Lecture 11 Programmed Cell Death III Methods to Study Autophagy Methods to study autophagy. Principles. (e.g. acidotropic dyes, LC3-II, p62, electrom microscopy ) Guidelines for the use and interpretation of assays for monitoring autophagy.
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Lecture 12 Methods to study Senescence Cellular Senescence Characteristic of senescent cells Replicative and induced senescence Senescent markers Senescence and cancer Senescence as target for chemotherapy Methods to study senescence
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Lecture 13 Future directions in PADS Future directions in PADS Translational oncology Implications of basic research on PADS Current trends in PADS
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Lecture 14 Concluding remarks Review of experimental data from the course Presentation by students
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LABORATORY FACILITIES Cell culture Microplate reading Microscopy (inverted, light and fluorescent) Electrophoresis and immunoblotting PCR
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LABORATORY TECHNIQUES Cell culture Short & Long Term Proliferation assays (CCK, RC0) Programmed cell Death Assays (DNA Laddering, Caspase activation Immunocytochemistry) Senescence Assays (beta-galactosidase assay) Western Blot (Apoptosis & Autophagy markers) PCR
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