| | Category | CELL | L13 | “Differential Response to Temozolomide in Patient-derived |
| | Glioblastoma Cells |
| | Abstract | Rationale |
| | Over 23,000 malignant tumors of the brain or spinal cord are |
| | diagnosed in 2016. Unfortunately, approximately 16,000 people die |
| | from their disease. Glioblastoma (GBM), the most common malignant |
| | brain tumor is often treated with temozolomide chemotherapy. There is |
| | an urgent need to identify the patients who will respond to |
| | temozolomide due to its side effects profile. The methylation of the |
| | promoter of the MGMT gene which codes for an enzyme critical in DNA |
| | repair is shown to be associated with temozolomide sensitivity in |
| | patients with GBM. Evaluation of the effect of temozolomide on patient- |
| | derived GBM cell lines with varying MGMT methylation status has the |
| | potential to provide additional insight on a specific drug’s activity ex- |
| | vivo. Future application of this approach will allow researchers to |
| | experiment on the patient’s tumor, not on the patient. |
| | |
| | Research question, hypothesis, goals, expected outcomes |
| | The purpose of this study is to determine the response to temozolomide |
| | therapy in three existing patient-derived GBM cell lines. The |
| | hypothesis is that the patient-derived GBM cell lines will mimic the |
| | known actual patient response to temozolomide, providing additional |
| | insight in the biology of the tumor. The expected outcome of this |
| | research is to show that response to temozolomide in the patient- |
| | derived GBM cell lines will be the same as in the GBM patient. |
| | |
| | Procedures |
| | Three patient-derived GBM cell lines (15-037, 14-015s, 14-104s) will be |
| | obtained from the cryo-preserved archives of patient-derived cell lines |
| | and tumor fragments. The samples in the archive were previously |
| | collected under an Institutional Review Board approved protocol and |
| | located in the Translational Neuro-Oncology Research Laboratory at |
| | the Karmanos Cancer Institute, Wayne State University School of |
| | Medicine, Detroit, MI. The three cell lines will be untreated or exposed |
| | to temozolomide (alkylating agent), or CH-223191 (aryl hydrocarbon |
| | antagonist). Cell proliferation assays will be assessed by the Vybrant |
| | MTT assay (Thermofisher). The GBM patient’s response to |
| | temozolomide will also be provided in a de-identified manner. |
| | |
| | Risk and Safety |
| | Tumor tissues and the resulting cell lines may pose a biological hazard |
| | with the risk of infection to exposed individuals. Temozolomide is an |
| | agent used for chemotherapy and is known to be cytotoxic. All tumor |
| | tissues and cell lines are handled in an annually re-certified biological |
| | safety cabinet. Personal protective equipment (gloves, lab coat, safety |
| | goggles) will be worn by all personnel performing the experiments. |
| | Using these established procedures, there is only a minimal risk of |
| | exposure. |
| | |
| | Data Analysis |
| | The numeric values obtained by the MTT assays will be analyzed for |
| | statistical significance of differences between control cells and treated |