| 1 | Introduction | At the first meeting, we will briefly outline the general requirements, the class schedule and administrative details. The instructors and students will introduce themselves. The instructors will then summarize basic aspects of cell cycle regulation and the DNA damage response network. We will introduce the different DNA damage checkpoints that evolved to protect cellular DNA from various sources of damaging agents. Also, commonly used model systems and standard techniques will be introduced and discussed. |
| 2 | The cell cycle | We will discuss the basic cell cycle machinery, using two classic papers describing the principles of cellular proliferation. We will learn about the different stages of the cell cycle and understand their specific characteristics. During this session we will also discuss the molecular mechanisms that make cell cycle progression dependent on extracellular cues (such as growth factors). |
| 3 | Cdk-regulation | We will discuss how the core cell cycle machinery is subject to regulation. Specifically, we will discuss the regulation of Cyclin-dependent kinases by phosphorylation and dephosphorylation. |
| 4 | Checkpoint regulation of the cell cycle | We will discuss the multiple means by which the DNA damage-activated checkpoints regulate cell cycle progression in response to DNA damage. Both transcriptional and post-translational responses will be discussed using the assigned landmark papers. |
| 5 | p53 regulation | We will discuss the important role of the prominent tumor suppressor gene p53 in the cellular responses to DNA damage. |
| 6 | The DNA damage checkpoint differs depending on cell cycle stage | We will dive deeper into the core mechanisms that control the cellular response to DNA damage. We will discuss different modes of regulation of the phosphatase Cdc25. Furthermore, we will analyze different types of repair strategies that depend on the cellular DNA content and thus on the cell cycle stage. |
| 7 | 'To die or not to die' - the decision between repair and apoptosis | We will discuss two different possible outcomes after exposure to genotoxic agents, controlled by p53. We will use two papers to show that both repair of the damaged genome and activation of programmed cell death can be initiated by similar signaling pathways. |
| 8 | Structural insights into the DNA damage response | We will discuss structural insights concerning the molecular machinery that controls the DNA damage response. We will focus on Rad50, a member of the MRN complex, as well as MDC1 and its interaction partner H2AX. These proteins are involved in the very early stages of the DNA damage response and together form the structures that serve as an anchoring platform for downstream signaling molecules. |
| 9 | Field trip: Visit to an MIT Biology laboratory | Now that we have gathered a solid background concerning checkpoint signaling, we will be introduced to the real methods. We will visit a molecular biology laboratory, focused on studying DNA damage and the subsequent checkpoint response. Typical checkpoint assays will be demonstrated. We will examine different samples of untreated or damaged material that are analyzed by FACS (Fluorescence Activated Cell Sorter) and by high-resolution fluorescence microscopy. We will analyze our data using the knowledge acquired in our prior sessions. |
| 10 | Defective DNA damage responses and cancer | We will learn about the importance of the DNA damage response by studying the impact of checkpoint failure due to mutations in critical checkpoint genes. We will discuss the consequences of loss of Chk1 and p53. Both papers use the mouse as a model organism. |
| 11 | Checkpoint-related syndromes | Defects in checkpoint signaling are associated with a number of human diseases. Besides spontaneous mutations in checkpoint genes, familial syndromes have been identified in which specific checkpoint genes are mutated. During this session, two such syndromes will be discussed and compared. |
| 12 | Treatment of disease based upon knowledge about the DNA damage response | We will discuss two recent papers that make use of our increasing understanding of DNA damage signaling to derive new therapeutic strategies that specifically target cancer cells rather than healthy cells. We will learn about the concept of personalized cancer therapy. |
| 13 | Final presentations | For the final presentations each student will be assigned a research article that describes the function of a critical checkpoint protein, which is related to a human disease. Students will discuss the key figures and summarize the molecular functions of the protein described in the paper. Furthermore, the students should briefly discuss the function of this protein within the global DNA damage network that was discussed throughout the course. Lastly, the students will be asked to explain the clinical features of the related human disease (mentioned in the research article), based on the molecular details that were described in the paper. The presentations are scheduled for 15 minutes (10 minutes talk [7-8 Microsoft® PowerPoint® slides], followed by a 5 minute discussion). The presentation will be given to the entire group. Students will be encouraged to actively participate in a five-minute discussion following each presentation. |