Distinct multiethnic genome-wide alterations in breast cancer using paraffin embedded samples: preliminary analysis

Abstract #2024 Background and Rational: Ethnic-specific disparities in breast cancer (BC) stage of presentation and survival rates are well documented. To further investigate possible ethnic-specific genetic contributions to these disparities, we are completing gene expression profiling studies in a multi-ethnic cohort consisting of thirty “Triple Negative” BC patients [10 each African-American (AA), Hispanic (His) and non-Hispanic white (Cauc) women] matched for age of diagnosis and hormone receptor status. The overall study aim is an increased understanding of the biological basis of ethnic-specific BC disparities, leading ultimately to individualized, ethnic-specific diagnostic and therapeutic approaches. Two immediate study goals are to demonstrate the utility of FFPE samples in obtaining consistent, reproducible data from gene expression arrays, and secondly, to identify differentially expressed genes between tumor and normal tissue that are common or unique among the three ethnic groups. Methods: Pathology specimens were freshly cut from FFPE blocks and marked by a pathologist as to normal vs. tumor tissue. RNA isolation, labeled cDNA preparation, and hybridization of tumor and normal cDNAs to a breast cancer focused gene expression microarray (Breast Cancer DSA Research Tool) was performed by Almac Diagnostics. Each patient was self-matched (tumor vs. normal tissue) for gene expression studies. Results: Using 36 matched tumor and normal FFPE samples from 18 patients, approximately 17516 transcripts were detected on the Breast Cancer DSA with intensity significantly greater than background. For normal and tumor tissue samples, 9399 and 10,296 transcripts respectively, were detected in all three ethnic groups. Importantly, a subset of transcripts (hundreds to one thousand) was detected in only one or two ethnic groups. Using two-way ANOVA (disease state and ethnicity), a subset of 6479 transcripts was identified with p-value less than 0.01 in the statistical test and was selected and further used in data quality control. Data QC indicated that patient samples clustered well with respect to both ethnicity and normal versus tumor tissue. Additional analytical methods included K-means 2-Dimensional clustering and Principal Component Analysis. From these analyses, we identified ethnic-specific expression patterns in the matched normal and tumor tissue samples. We are completing these studies by increasing sample size and matching for stage of diagnosis, mapping clusters of differentially-expressed genes in pathway analysis, and validation by real-time PCR. In the longer term, DNA copy number variation (CNV) and chromosomal alterations will be investigated by high density arrays. Summary: These preliminary analyses shows that high quality gene expression data can be generated from FFPE samples, and that ethnic specific gene expression differences can be detected in tumor and matched normal breast tissue samples across ethnic groups. Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2024.