The goal of this project was to further characterize select genes in triple negative breast cancers (TNBC) in an effort to character the cancers and ultimately understand the pathogenesis of the cancers. A primary goal was to identify genes that might define the heterogeneity of the disease. Previous data in our laboratory identified Interleukin 32 (IL-32) gene as differentially expressed in a particular subpopulation of TNBC compared to non-tumor and luminal cancers. Further studies showed that there appeared to be 2 groups of TNBC defined by their levels of IL32 gene expression; those defined as "IL32 High compared to IL32 Low" gene expression. Using T-test analyses we were able to identify genes of interest that were differentially expressed between the two populations. Utilizing Gene Ontology program analysis, a subset of the genes were found to be immune-related genes. From these data, we suggest that IL32 and the related immune panel of genes define a subpopulation of TNBC and might serve to define and/or drive the particular breast cancer genotype. Validation of our gene set as so called drivers in sustaining the TNBC genotype are beyond the scope of this project; instead, the aim of the current project is to validate the differential expression of these gene in a subpopulation of TNBC. These experiments should allow us to characterize and better understand TNBC. Towards this aim, we will (a) validate the gene expression levels of a small panel of immune-related (and other) genes in the cell lines previously identified in IL32-high-TNBC and IL32-low-TNBC, and (b) select 'a promising' gene candidate and examine its protein expression level in tissue microarray clinical samples. As TNBC are known to be a heterogeneous subtype, our experiments should help in further characterizing TNBC and for the first time begin to address the possible utility of our gene panel in these cancers.