Project Details

Description

Breast Cancer 1000 Project 2012-2013 Breast Cancer 1000 Project 2012-2013 Resistance to therapy in breast cancer patients is a serious therapeutic problem and major efforts are underway to understand underlying mechanisms. Resistance can be either intrinsic or acquired. Our single cell sub-cloning demonstrated that many MCF7 cells are resistant de novo to tamoxifen, even without selection under pressure. In this proposal we will continue characterizing tamoxifen sensitive and resistant MCF7 subclones at genomic and proteomic level. Data obtained will be aligned across methods and genes and pathways that play a role in tamoxifen resistance will be validated in functional studies. We will continue working in the characterization of the different p53 mutant stable MCF10A cells to better understand the biological importance of this important tumor suppressor gene. We will also performed a functional screen to analyze the importance of mutations in other genes and in this form find alternative targets that may offer better alternative for breast cancer patients. We propose the following specific aims for the 2012-2013 funding period: Continue with the characterization of tamoxifen sensitive and resistant breast cancer subclones. 1. Genomic analysis of tamoxifen sensitive and resistant MCF7 subclones. i. Finish the computational/statistical analysis of G11 and use the HME1 for data normalization. ii. Identify the genomic differences across sensitive and resistant cells iii. Perform a pathway analysis to identify relevant pathways for tamoxifen resistance. iv. Validate relevant genes involved in resistance by performing functional genomic analyses through ectopic expression or silencing of the gene of interest to confirm their role in changing phenotype on cells. For example if gene X is lost in resistant cells, we will knock it down in sensitive cells. If it plays a role in resistance, cells should grow better in presence of tamoxifen. v. Prepare manuscript for publication. vi. If possible similar genomic approach will be perform in tumor samples from tamoxifen resistant patients. 2. Proteomic analysis of the relative abundance of proteins of tamoxifen sensitive and resistant cells. i. Finish with data analysis and perform a pathway analysis. ii. Validate the most interesting proteins by western blot. iii. This data will complement the gene expression profiling and genome analysis of subclones. In conjunction with all these data we will be able to have a global picture of tamoxifen resistance. iv. Prepare manuscript for publication. 3. Effect of co-drivers with TP53 mutation in breast cancer progression. i. Evaluate induction of EMT staining by IF and WB in all MCF10A-mt stable, WT and NULL TP53 cells ii. Evaluate differences in cell proliferation and escape of apoptosis. iii. Evaluate differences in migration, invasion and anoikis. iv. Prepare manuscript for publication. v. Perform functional screen to evaluate the effect of some co-drivers in hallmarks of cancer (EMT, apoptosis escape and anoikis) 4. Continue supporting the community by providing plasmids from our human collections (BC1000 and Kinases).
StatusFinished
Effective start/end date10/1/129/30/13

Funding

  • Breast Cancer Research Foundation: $233,964.00

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