title: Novel tumor sampling strategies to enable microarray gene expression signatures in breast cancer: a study to determine feasibility and reproducibility in the context of clinical care.
abstract: Feasibility and reproducibility of microarray biomarkers in clinical settings are doubted because of reliance on fresh frozen tissue. We sought to develop and validate a paradigm of frozen tissue collection from early breast tumors to enable use of microarray in oncology practice. Frozen core needle biopsies (CNBx) were collected from 150 clinical stage I patients during image-guided diagnostic biopsy and/or surgery. Histology and tumor content from frozen cores were compared to diagnostic specimens. Twenty-eight patients had microarray analysis to examine accuracy and reproducibility of predictive gene signatures developed for estrogen receptor (ER) and HER2. One hundred twenty-seven (85%) of 150 patients had at least one frozen core containing cancer suitable for microarray analysis. Larger tumor size, ex vivo biopsy, and use of a new specimen device increased the likelihood of obtaining adequate specimens. Sufficient quality RNA was obtained from 90% of tumor cores. Microarray signatures predicting ER and HER2 expression were developed in training sets of up to 363 surgical samples and were applied to microarray data obtained from core samples collected in clinical settings. In these samples, prediction of ER and HER2 expression achieved a sensitivity/specificity of 94%/100%, and 82%/72%, respectively. Predictions were reproducible in 83-100% of paired samples. Frozen CNBx can be readily obtained from most breast cancers without interfering with pathologic evaluation in routine clinical settings. Collection of tumor tissue at diagnostic biopsy and/or at surgery from lumpectomy specimens using image guidance resulted in sufficient samples for array analysis from over 90% of patients. Sampling of breast cancer for microarray data is reproducible and feasible in clinical practice and can yield signatures predictive of multiple breast cancer phenotypes.
authors.affiliation: Department of Surgery, Duke University Medical Center, Durham, NC 27710, USA. christopher.tebbit@duke.edu
authors.forename: Christopher L
authors.initials: CL
authors.lastname: Tebbit
authors.forename: Jun
authors.initials: J
authors.lastname: Zhai
authors.forename: Brian R
authors.initials: BR
authors.lastname: Untch
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Ellis
authors.forename: Holly K
authors.initials: HK
authors.lastname: Dressman
authors.forename: Rex C
authors.initials: RC
authors.lastname: Bentley
authors.forename: Jay A
authors.initials: JA
authors.lastname: Baker
authors.forename: Paul K
authors.initials: PK
authors.lastname: Marcom
authors.forename: Joseph R
authors.initials: JR
authors.lastname: Nevins
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Marks
authors.forename: John A
authors.initials: JA
authors.lastname: Olson
mesh_terms: D014408:Biomarkers, Tumor / Q000032:analysis* / Q000235:genetics; D044963:Biopsy, Fine-Needle / Q000379:methods*; D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology / Q000601:surgery*; D005240:Feasibility Studies; D005260:Female; D005629:Frozen Sections; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D009367:Neoplasm Staging; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics; D011960:Receptors, Estrogen / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D025321:Surgery, Computer-Assisted
chemical_list: D014408:Biomarkers, Tumor; D018719:Erb-b2 Receptor Tyrosine Kinases; D011960:Receptors, Estrogen