Juan Sebastian Yakisich  
Preclinical  Anticancer Drugs Screening (PADS) Based on Novel Models of Cancer BiologyJuan Sebastian Yakisich, MD, PhD

JSY - OncoDrugs



Instructor: Juan Sebastian Yakisich

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.


e-mail: yakisich@gmail.com






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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.









Selected scientific articles will be provided



Cell  Biology, Molecular Biology,  Chemistry,



Lectures   Laboratory

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.



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.



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.



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)



Lecture 5

Cancer Biology I

Basic Aspects of Cancer Cells

Characteristics of cancer cells.



Mechanisms of drug-resistance.



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.



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).



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)



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



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



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.



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



Lecture 13

Future directions in PADS

Future directions in PADS

Translational oncology

Implications of basic research on PADS

Current trends in PADS



Lecture 14

Concluding remarks

Review of experimental data from the course

Presentation by students




Cell culture

Microplate reading

Microscopy (inverted, light and fluorescent)

Electrophoresis and immunoblotting





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)







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