| 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 |