Abstract B5: Sequence analysis of HSP90-encoding genes in breast tumor tissue identifies a novel 9 bp deletion in HSP90AB1

The 90 kDa heat-shock protein (HSP90) functions as a ubiquitous cellular chaperone in cells, and is required for the function and stability of many client proteins such as ERBB2, EGFR, BCR-ABL, MET, androgen and estrogen receptors, BRAF and HIF-1α. 1 There are four primary genes that encode for HSP90 in humans: HSP90AA1 and HSP90AB1 (cytosolic), HSP90B1 (endoplasmic reticulum) and TRAP1 (mitochondrial). Increased HSP90 expression has been associated with poor prognosis in cancer, and inhibiting HSP90 can lead to degradation of many oncogenic client proteins via the proteasome pathway. 1 HSP90 has been the subject of intense drug discovery activities, with several therapeutics under clinical investigation. 2 However, the data on single nucleotide polymorphisms (SNPs) and potential somatic mutations within the HSP90-encoding genes in tumor tissue is limited. Target allelic variation can alter drug binding and therapeutic response, as demonstrated by Iressa™9s reported specific efficacy in EGFR-mutant non-small cell lung cancer patients. 3 Therefore, it is critical to understand target allelic variance as part of a drug development program. To facilitate drug development efforts around HSP90, we investigated allelic variation within the coding regions of HSP90AA1 (second isoform) and HSP90AB1 genes in 49 formalin-fixed paraffin- embedded human breast tumor samples. Sample SNP data were compared to control data from the NCBI Entrez SNP database. We observed four synonymous changes within HSP90AA1, and two synonymous changes within HSP90AB1. Intriguingly, we also observed a heterozygous 9-base deletion in exon 1 of HSP90AB1 within one breast tumor patient sample. This deletion results in an alanine>valine substitution at amino acid 22 and a loss of amino acid 23-25 (glutamine, leucine, methionine). The deleted region includes one turn of a 3-turn alpha helix, located within the N-terminal domain structure of the HSP90AB1 protein. Full details of HSP90 allelic variance within breast tumor tissue, and a discussion of the potential functional significance of the deletion, will be provided. References: 1. Pearl LH, Prodromou C, Workman P. The HSP90 molecular chaperone: an open and shut case for treatment. Biochem J. 2008;410:439-53. 2. Hahn JS. The HSP90 chaperone machinery: From structure to drug development. BMB Reports. 2009;42(10):623-30. 3. Sequist LV, Bell DW, Lynch TJ, Haber DA. Molecular predictors of response to epidermal growth factor receptor antagonists in non- small-cell lung cancer. J Clin Oncol. 2007;25(5):587-95. Citation Information: Clin Cancer Res 2010;16(14 Suppl):B5.