Characterization of Necroptosis in Breast Cancer and Melanoma for Oncolytic Vaccinia Potential
Description
With the advent of precision medicine, oncologists aim to target tumors that do not respond well to conventional treatment. One such therapy is oncolytic virotherapy, a treatment reliant on viral replication for tumor specific killing. Downregulation of the proteins RIP3 kinase, DAI or MLKL can result in a nonfunctional programmed necroptotic cell death pathway, common amongst breast cancer and melanoma. Vaccinia virus (VACV) mutants with a nonfunctional E3 protein are able to selectively replicate in necroptosis deficient cells but not in necroptosis competent cells, making them potential candidates for oncolytic virotherapy. In order to establish the efficacy and selectivity of this treatment, an accurate tumor model is required. Eight established breast adenocarcinomas and two established melanomas were selected as potential candidates, both human and murine. A pan screening method for necroptosis was established utilizing western blot analysis for expression of aforementioned proteins following various induction methods such as IFN α or VACV infection. In addition, live cell imaging after treatment with tumor necrosis factor (TNFα) and the pan-caspase inhibitor zVAD-fmk was used as a method to visualize necroptosis pathway functionality. Based on these results, cell lines will be selected and modified to create a breast cancer model with cells that are syngeneic, differing only in expression of either RIP3. VACV can be tested for tumor volume reduction in these models to ask if RIP3 expression affects efficacy of mutant VACV as an oncolytic virus.
Date Created
The date the item was original created (prior to any relationship with the ASU Digital Repositories.)
2019-05
Agent
- Author (aut): Kumar, Aradhana
- Thesis director: Jacobs, Bertram
- Committee member: McFadden, Grant
- Committee member: Borad, Mitesh
- Contributor (ctb): School of Life Sciences
- Contributor (ctb): School of Life Sciences
- Contributor (ctb): Barrett, The Honors College