Ewing's sarcoma is a childhood/young adult cancer. Unfortunately, many Ewing's sarcoma cancer patients are first diagnosed when the cancer cells are already spreading throughout the body, and these patients have a very poor prognosis. Therefore, identifying novel means to improve the effectiveness of the standard treatment of this cancer, a cocktail of drugs that includes etoposide, would be of immediate benefit. This proposal will provide the basis for novel strategies in the treatment of human cancer. Towards this goal, our work has been to identify the mechanisms that a normal (non-cancer) cell uses to survive exposure to an agent such as etoposide. The first part of our hypothesis is that we can identify novel human etoposide survival mechanisms from a Drosophila RNAi screen. We plan to use this knowledge to determine whether any etoposide survival mechanisms are altered in Ewing's sarcoma cancer cells. The second part of our hypothesis is that we can target etoposide survival mechanisms to restore or improve the effectiveness of etoposide therapy depending on what has been altered in the cancer cell. To achieve these goals, we will build from our currently established methods of identifying survival mechanisms in cells from a model organism;Drosophila cells. Once these genes are determined in Drosophila cells, we can test whether the same genes contribute to survival in mammalian cells. Indeed, we already successfully used this strategy to identify novel alkylation survival genes and pathways in mouse and human cells and demonstrated that pathways critical in Drosophila are also involved in mammalian damage response. The advantage of using fruit fly cells is that they share the same processes to survive exposure to an agent such as etoposide as human cells and we can efficiently, rapidly and cost-effectively ask whether removing any one gene alters the ability of a cell to survive etoposide treatment. This provides us with the ability to identify novel genes/mechanisms involved in cell survival. We will then demonstrate that the survival processes we identify in fruit fly cells are also used in mammalian cells. For this we plan to use human Ewing's sarcoma cells.
In Aim 1, we propose to leverage our strategy, first completing the validations for a genomic survival screen to etoposide, identifying survival mechanisms, and then we will determine which of these mechanisms are altered in Ewing's sarcoma cells in a EWS/FLI1 dependent manner and which can be targeted to sensitize these cells to etoposide.
In Aim 2, we will directly test the functionality of already identified survival mechanisms (and any identified from Aim 1) in a set of Ewing's sarcoma cancer cells, demonstrating any underlying heterogeneity in these cancers, and whether targeting these different pathways effects etoposide sensitivity in these cells. We are uniquely positioned to carry out the proposed work with the necessary infrastructure, collaborators and technical expertise, as demonstrated by our publications and our preliminary data that supports our hypothesis. Ultimately, our goal is to improve etoposide treatment of Ewing's sarcoma.

Public Health Relevance

DNA damage is a normal consequence of life that is often implemented in the treatment of cancers, however, the resultant cellular damages must be counteracted in normal cells in order to maintain cellular and organism fitness. Alteration in one of these countermeasures is one basis for cancer cells to become resistant to treatment, making the treatment ineffective. Here we propose to discover mechanisms used to counteract DNA damage caused by the cancer therapy agent, etoposide, and target these mechanisms in conjunction with etoposide to augment its effectiveness in treating Ewing's sarcoma.

National Institute of Health (NIH)
National Cancer Institute (NCI)
Research Project (R01)
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Basic Mechanisms of Cancer Therapeutics Study Section (BMCT)
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Pelroy, Richard
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University of Texas Health Science Center
Schools of Medicine
San Antonio
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Zanotto-Filho, Alfeu; Dashnamoorthy, Ravi; Loranc, Eva et al. (2016) Combined Gene Expression and RNAi Screening to Identify Alkylation Damage Survival Pathways from Fly to Human. PLoS One 11:e0153970
Hsiao, Tzu-Hung; Chiu, Yu-Chiao; Hsu, Pei-Yin et al. (2016) Differential network analysis reveals the genome-wide landscape of estrogen receptor modulation in hormonal cancers. Sci Rep 6:23035
Zanotto-Filho, Alfeu; Masamsetti, V Pragathi; Loranc, Eva et al. (2016) Alkylating Agent-Induced NRF2 Blocks Endoplasmic Reticulum Stress-Mediated Apoptosis via Control of Glutathione Pools and Protein Thiol Homeostasis. Mol Cancer Ther 15:3000-3014
de Araujo, Patricia Rosa; Gorthi, Aparna; da Silva, Acarizia E et al. (2016) Musashi1 Impacts Radio-Resistance in Glioblastoma by Controlling DNA-Protein Kinase Catalytic Subunit. Am J Pathol 186:2271-8
Chiu, Yu-Chiao; Wu, Chin-Ting; Hsiao, Tzu-Hung et al. (2015) Co-modulation analysis of gene regulation in breast cancer reveals complex interplay between ESR1 and ERBB2 genes. BMC Genomics 16 Suppl 7:S19
Lei, Chengwei; Ruan, Jianhua (2013) A novel link prediction algorithm for reconstructing protein-protein interaction networks by topological similarity. Bioinformatics 29:355-64
Lei, Chengwei; Tamim, Saleh; Bishop, Alexander Jr et al. (2013) Fully automated protein complex prediction based on topological similarity and community structure. Proteome Sci 11:S9
Karia, Bijal; Martinez, Jo Ann; Bishop, Alexander J R (2013) Induction of homologous recombination following in utero exposure to DNA-damaging agents. DNA Repair (Amst) 12:912-21
Jahid, Md Jamiul; Ruan, Jianhua (2012) A Steiner tree-based method for biomarker discovery and classification in breast cancer metastasis. BMC Genomics 13 Suppl 6:S8
Doderer, Mark S; Anguiano, Zachry; Suresh, Uthra et al. (2012) Pathway Distiller - multisource biological pathway consolidation. BMC Genomics 13 Suppl 6:S18