Investigational Agents to Target Specificity Protein1 Transcription Factor and Survivin for Inhibiting Medulloblastoma Cells Growth

Abstract

Abstract Presenter: Serena Bao Authors: Serena Bao, Sachi Kishinchandani, Briggs Lambring, Umesh Sankpal, Iloani Nwamaka, Riyaz Basha Title: Investigational Agents to Target Specificity Protein1 Transcription Factor and Survivin for Inhibiting Medulloblastoma Cells Growth Background: Medulloblastoma (MB) is a brain cancer predominantly arising in children. It accounts for 20% of all childhood brain tumors. The treatment for MB includes a combination of surgery and radiation therapy. Although around 70% of the patients have shown remission with treatment today, these therapies are associated with significant morbidity, especially in the youngest patients. Therefore, widespread interest is shown to develop more successful treatments. One potential target of cancer treatment is a protein known as survivin. Survivin inhibits cell death and is upregulated in most cancers, including MB. A transcription factor known as Sp1 upregulates survivin by binding to its promoter region. Therefore, suppressing Sp1 activities indirectly down-regulates survivin and can serve as a target for MB therapy. Our aim is to find and test appropriate investigational agents that inhibit Sp1, thereby inhibit survivin and cancer cell growth. In addition, we want to use an in silico analysis to determine the expression of Specificity protein1 (Sp1) and survivin in MB patients, and see how it correlates with the survival of these patients. It has been previously determined that the investigational agents Tolfenamic acid (TA) and its derivative copper-tolfenamic acid (CuTA) are effective at inhibiting Sp1 in some cancer cells. In this project, we investigated the growth inhibitory effect of TA and CuTA using DAOY cell line. Method: DAOY cell lines (purchased from a commercial source, American Type Culture Collection, Manassas, Virginia; IBC-2016-0038) were cultured and seeded (2,000 cells per well) in a 96 well plate. Cells were treated using increasing concentrations (0, 10, 20, 40, 80 µM) of TA or Cu-TA. After a 48-hour incubation, the viability of the sample was measured via a luminance assay using the CellTiterGlo. Dose curves were generated, and the dose required to inhibit 50% of the viability (IC50 value) was determined using Graphpad Prism Software. Data from the R2 genomics visualization platform was used to generate Kaplain-Meier curves. The presented curves compare survival probabilities in Medulloblastoma patients with high vs. low expression of Sp1 and survivin. Results: The in silico analyses using R2 genomics visualization platform demonstrated MB patients who express higher levels of Sp1 is associated with low survival time(p = 0.033). Similarly, MB patients with higher levels of survivin expression show poor prognosis (p = 0.049) As per the IC50 values, Cu-TA is 3.6 times more effective against MB cells without affecting non-cancerous cells than TA Conclusion: Higher expression of Sp1 or survivin is associated with low survival time in medulloblastoma patients. Both TA and Cu-TA are inhibiting DAOY cell growth, however, Cu-Ta is more effective than TA against MB cell line. With increased resistance to standard therapies, TA and Cu-TA potentially enhance the therapeutic efficacy of chemotherapy and radiation.