Enzyme-Catalyzed Mutation in Breast Cancer
Abstract
Breast cancer development requires multiple mutations, some inherited and others acquired somatically, but the molecular causes of these mutations remain largely unknown. We hypothesize that the DNA cytosine deaminase APOBEC3B contributes directly to breast tumorigenesis by making both mutations and epigenetic alterations, that ultimately endow tumor cells with selective growth advantages. Aim 1 tests the genetic hypothesis by depleting endogenous APOBEC3B levels in representative breast cancer cell lines and, in comparison to control shRNA expressing cell lines, determining the distribution and pattern mutations by deep-sequencing and asking whether APOBEC3B levels impact the rate of therapy resistance in well-established xenograft mouse models. The first phases of these experiments have been completed and DNA samples have been prepared for deep sequencing. Aim 2 tests the epigenetic hypothesis by depleting endogenous APOBEC3B levels in representative breast cancer cell lines and by overexpressing APOBEC3B or a catalytically inactive control in normal breast epithelial cell lines and quantifying global and local changes in MeC content. These cell lines have been engineered as proposed and preliminary data analyses are underway. This work is significant because it will delineate a major source of mutations and epigenetic changes in breast cancer, which paves the way for new diagnostic/prognostic tests and methods to treat breast cancer by preventing the activity of this enzyme.
Document Details
- Document Type
- Technical Report
- Publication Date
- Aug 01, 2014
- Accession Number
- ADA613711
Entities
People
- Reuben S Harris
Organizations
- University of Minnesota