We describe 2 karyotypically male infants with terminal deletion of 10q and mental retardation, multiple phenotypic anomalies and abnormal genitalia. One [karyotype 46,XY, del(10)(q26.1)] had female external genitalia; the other [karyotype 46,XY,-10,+der(10)t (10;16)(q26.2;q21)] had an intersex phenotype. Of 8 males previously reported with terminal 10q deletion as the major or only cytogenetic abnormality, 2 had an intersex phenotype, and the others all had combinations of cryptorchidism, micropenis, and hypospadias. Terminal 10q deletions appear to be strongly associated with abnormal male genital development, and should be specifically searched for in the cytogenetic workup of such cases.

#3002 Introduction: Aberrations of chromosome 17 (aneusomy) are common in breast cancer and therefore have a critical impact on the assessment and reporting of HER2 gene amplification in a significant sub-set of cases. There is an ongoing debate as to the proportion of cases for which assessment of chromosome 17 copy number may be important. According to current guidelines1,2, amplification of HER2 is considered to be a HER2/chromosome 17 ratio ≥2.0 and a ratio <2.0 is regarded as non-amplified. For HER2 gene copy number assays, it has been assumed that copy numbers of >6.0 reflect amplification and a result of <4.0 HER2 gene copies per nucleus is always associated with lack of amplification; cases with 4-6 copies per cell are thought to require validation by testing of a parallel section for chromosome 171,2. To our knowledge, this assumption has not been verified experimentally.
 Methods: HER2 and chromosome 17 were measured by dual color FISH in 1711 breast cancer samples referred to the authors laboratories between 2000-2008. Using HER2 copy number and chromosome 17 data the impact of chromosome 17 testing upon accuracy of diagnosis of gene amplification was assessed.
 Results: At a HER2 copy number of 2 to <3, 16 of 488 cases (3.3%) had HER2 amplification; and at a copy number of 3 to <4, 32 of 195 cases (16.4%) were amplified. The proportion of cases with HER2 amplification increased considerably at HER2 copy numbers of 4 to <7: 50.0% at 4 to <5; 67.5% at 5 to <6, and 77.3% at 6 to <7. Virtually all cases were amplified at HER2 copy numbers of ≥7.
 Conclusion: Werecommend that all cases with observed HER2 copy numbers of 2 to 7 should also be analyzed for chromosome 17 in order to accurately determine HER2 gene amplification. This would require analysis of chromosome 17 in 48.3% of all breast cancer cases based upon the study population. Current guidelines1,2 recommend chromosome 17 measurement only in cases with a HER2 copy number of 4 to <6, which represents only 6.6% of all breast cancer cases in the current study. Although single color ISH is becoming more widely used with the availability of CISH, the importance of chromosome 17 measurement cannot be ignored. It is essential that HER2 testing is of high quality, so that optimal patient management can be provided.
 1. Ellis IO, Bartlett J, Dowsett M et al: Updated recommendations for HER2 testing in the UK. J.Clin.Path. 57(3), 233-237 (2004).
 2. Wolff AC, Hammond ME, Schwartz JN et al: American Society of Clinical Oncology/College of American Pathologists Guideline Recommendations for Human Epidermal Growth Factor Receptor 2 Testing in Breast Cancer. Journal of Clinical Oncology 25(1), 118-145 (2007). Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 3002.

#45 Background: The role of HER2 and Topoisomerase IIα (TOPO2α) as predictors of anthracycline sensitivity remains controversial. To address this we conducted an analysis of HER2, Topoisomerase IIα amplification (TOPO2α-amp) and deletion (TOPO2α-del), chromosome 17 polysomy (C17) and Ki67 as prognostic and predictive markers in the UK National Epirubicin Adjuvant Trial (NEAT) which compared classical CMF with Epirubicin followed by CMF (NEJM 2006;355:1851-62).
 Methods: TMAs constructed with 1638/2021 tumors from patients in the NEAT study were analysed for HER2/TopoIIα gene alterations, C17 polysomy, and Ki67. Log-rank analyses explored the prognostic value of markers on relapse-free survival (RFS) and overall survival (OS). Cox-regression models tested independent prognostic value on RFS and OS in the presence of treatment, age, type of surgery, tumor size, nodal status, ER status and grade, and marker x treatment interactions for RFS and OS.
 Results: Of 1625 NEAT samples analysed, 806(50%) received anthracycline-containing treatment; 985(61%) were ≤50 years old; 1114(69%) had nodal involvement; 791 (49%) were ER+ve; 961(59%) had G3 tumours; and 893 (56%) had tumour size >2cm. 19% were HER2-amp, 9% were TOPO2α-amp, 9% were TOPO2α-del, 18% were polysomic for C17 and 62% had high Ki67 (>13.0%). HER2-amp and TOPO2α-del were significant poor prognostic factors (P<0.001) for RFS and OS, and were independent variables in multivariate analyses. Other factors did not reach significance at the 1% level. No significant treatment interaction with anthracyclines was seen for HER2-amp (OS p=0.25, RFS p=0.51). Polysomy C17 was not prognostic but exhibited a significant treatment interaction with anthracyclines (RFS p=0.02, HRs 0.96 95%CI 0.73-1.26 vs 0.56 95%CI 0.33-0.97, OS p=0.06, HRs 0.97 95%CI 0.72-1.30 vs 0.60 95%CI 0.33-1.08). Patients with C17 polysomic tumours had significantly greater benefit from anthracyclines.
 Conclusions: Analysis of samples from the NEAT trial, which for the first time included HER2, TOPO2α, Ki67 and C17, strongly suggest that the most powerful predictor of benefit from adjuvant anthracyclines is chromosome 17 polysomy, perhaps as a marker of chromosomal instability, with a weak effect for HER2-amp. No effect was observed for differing TOPO2α status. Given the lack of consistency seen in previous studies with respect to HER2 and TOPO2α, we suggest that polysomy C17 could be the unifying predictive marker of anthracycline sensitivity, particularly as these data are confirmed by data from a separate meta-analysis of MA5 and BR9601 trials (SABCS2008). Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 45.

Background: The Tamoxifen and Exemestane Adjuvant Multinational (TEAM) trial included prospectively planned biomarker studies to identify prognostic and predictive biomarkers for patients receiving endocrine therapy. Quantitative IHC data for ER/PgR (Can Res 69:83S, SABCS2008), HER2, HER3 and Ki67 was available for the current analysis relative to outcome of estrogen receptor–positive (ER+) early postmenopausal breast cancer (BC) patients treated with exemestane versus tamoxifen.Patients & Methods: Pathology blocks from 4598 TEAM patients were collected and tissue microarrays constructed. Quantitative analysis of hormone receptors (HER2/3) by conventional IHC, and image analysis derived continuous scores for Ki67/ER/PgR were analyzed relative to disease-free survival and treatment on an intent to treat basis using survival data for the first 2.75 years of the TEAM trial. Data on HER2FISH and EGF Receptor IHC will be presented.Results: Of 4595 eligible cases samples received, 16 were excluded, 271 had incomplete biomarker data, leaving 4308 patients for the final biomarker analysis. 1275 (30%) cases were HER2/3 positive.A significant treatment by marker effect was observed for exemestane versus tamoxifen with HER2/3 negative cases deriving benefit from aromatase inhibitor treatment (HER2/3-ve HR=0.69 95%CI, 0.53-0.88; HER2/3 pos HR, 1.13; 95%CI, 0.82–1.55; p=0.016 for interaction in multivariate analysis). By conventional and STEPP analysis no predictive effect of Ki67 was observed. In multivariate regression analysis increased HER2 expression (P=0.0001) decreased PgR expression (P<0.0001) and increased percentage of Ki67 positive cells (P=0.004) as continuous IHC variables were independently prognostic as were size (P=0.0001), nodal status (P<0.0001), grade (P=0.03) and age (P<0.0001).Conclusion: Multiple biological parameters (HER2/PgR/Ki67) are independently prognostic in ER+ve early postmenopausal BC. Modelling will be explored to derive prognostic and potentially predictive biomarker signatures for application in BC. Preferential exemestane versus tamoxifen treatment benefit was seen in HER2/3 negative cases, whilst HER2/3 positive cases had a poor prognosis in this population receiving hormonal therapy (suggesting resistance to endocrine therapy), and no evidence of benefit from AIs versus tamoxifen. Type I receptor tyrosine kinases may identify breast cancers with relative resistance to all forms of endocrine therapy. Whilst Ki67 alone was not predictive of benefit from Ais, Ki67, HER2 and PgR were independent prognostic variables and modelling of predictive/prognostic effects may further inform treatment selection in early postmenopausal breast cancer. Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 75.

Background: The Tamoxifen and Exemestane Adjuvant Multinational (TEAM) trial included prospectively planned biomarker studies to identify predictive biomarkers for patients receiving endocrine therapy. Quantitative IHC data for ER/PgR, HER1, HER2, HER3 and FISH analysis of HER2 in all cases was available for the current analysis relative to outcome of estrogen receptor-positive (ER+) early breast cancer patients treated with exemestane versus tamoxifen. Patients & Methods: Pathology blocks from 4598 TEAM patients were collected and tissue microarrays constructed. Quantitative analysis of receptors (HER1/2/3) by conventional IHC, and FISH (for HER2 only) were analysed relative to disease-free survival and treatment on an intent to treat basis using survival data for the first 2.75 years of the TEAM trial. Results: Of 4595 eligible cases samples received, 16 were excluded, and 4010 had complete biomarker data for all markers (HER1/HER2 & HER3) for the final biomarker analysis, 3.5% were HER1 positive, 13% HER2 positive & 21% HER3 positive. 1248 (31%) cases were HER1or2or3 positive (HER1-3+ve). HER1-3 positivity was associated with poor outcome (HR=1.6 95%CI=1.3-2.0). In HER1-3 negative patients the hazard ratio (for risk of relapse on exemestane versus tamoxifen in the first 2.75 years) was 0.68 (95% CI = 0.52-0.89), for the HER1-3 positive cases the hazard ratio was 1.14 (95% CI = 0.83-1.56) with a significant treatment by marker interaction (HR=1.68 95%CI=1.1-2.5; p=0.0014 in multivariate analysis). Trends for similar effects were seen for HER1 negative (-ve) vs HER1 positive (+ve) (HRs 0.80, 95%CI=0.65-0.99 vs 1.63 95%CI=0.74-3.59), HER2-ve vs HER2+ve (HRs 0.71, 95%CI=0.57- 0.9 vs 1.69 95%CI=108-2.63) and HER3-ve vs HER3+ve (HRs 0.78 95%CI=0.62-0.99 vs 1.04 95%CI=0.67-1.62) breast cancers. Conclusion: Preferential exemestane versus tamoxifen treatment benefit was seen in HER1/2/3 negative cases, whilst HER1/2/3 positive cases had a poor prognosis in this endocrine treated population (suggesting a degree of resistance to endocrine therapy), and no evidence of additional benefit from AIs versus tamoxifen. These three Type I receptor tyrosine kinases appear to identify breast cancers with relative resistance to all forms of endocrine therapy. This prospectively planned and powered treatment by marker analysis provides high level scientific evidence which may assist clinicians and patients in determining optimal AI schedules for women with early breast cancer. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr S2-4.