General, the pCR rates were similar in wild-type and = .323). the dual anti-HER2 blockade. The integrated analysis of gene expression and copy number data demonstrated that a 50-gene signature specifically predicted the lapatinib-induced pCR. Conclusion. mutations seem to identify patients who are less likely to benefit from dual anti-HER2 inhibition. p95-HER2 and markers of phosphoinositide 3-kinase pathway deregulation are not confirmed as markers of different sensitivity to trastuzumab or lapatinib. Implications for Practice: HER2 is currently the only validated marker to select breast cancer patients KRN 633 for anti-HER2 treatment; however, it is becoming evident that HER2-positive breast cancer is a heterogeneous disease. In addition, more and more new anti-HER2 treatments are becoming available. There is a need to identify markers of sensitivity to different treatments to move in the direction of treatment personalization. This study identified mutations as a potential predictive marker of resistance to dual anti-HER2 treatment that should be further studied in breast cancer. mutation has a prognostic impact in advanced HER2-positive disease [11, 12]. The results of the CHER-LOB (Chemotherapy, Herceptin and Lapatinib in Operable Breast Cancer) study showed that dual HER2 blockade with trastuzumab and lapatinib combined with chemotherapy resulted in a significantly increased pCR rate compared with single HER2 blockade with either lapatinib or trastuzumab plus chemotherapy [13]. In this paper, we report the results of the preplanned translational biomarker program of Rabbit Polyclonal to LW-1 the CHER-LOB study. Methods Clinical Platform CHER-LOB is a phase II randomized multicenter trial in which 121 patients with primary HER2-positive breast cancer were randomized to receive preoperative chemotherapy with weekly paclitaxel for KRN 633 12 weeks followed by 4 weekly courses over 3 weeks of the FEC regimen (fluorouracil, epirubicin, and cyclophosphamide) plus either trastuzumab (arm A), lapatinib (arm B), or the combination of trastuzumab and lapatinib (arm C). The trial design; eligibility criteria; statistical analysis; and clinical results, including response, surgery outcomes, and treatment safety, have been described in detail elsewhere [13]. Briefly, the main inclusion criteria included a diagnosis of breast cancer stage II to IIIA, HER2 positivity according to the local laboratory (immunohistochemistry [IHC] 3+ or fluorescence in situ hybridization [FISH] amplification), and no prior therapy for breast cancer. The translational biomarker program included the central reassessment of HER2 status, protein biomarker evaluation (p95-HER2, PTEN, phosphorylated AKT [pAKT], Ki67, terminal deoxynucleotidyl transferase dUTP nick end labeling [TUNEL]), the assessment of gene expression profile and copy number (CN) variations, and the study of somatic mutations of Mutation Analysis Three 5-m FFPE sections of a primary lesion containing at least 50% tumor cells were deparaffinized and incubated in lysis buffer with proteinase K (50 mM Tris, 1 mM EDTA, 05% TWEEN 20) at 56C overnight. Genomic DNA was extracted with QIAmpl DNA Mini Kit (Qiagen, Valencia, CA, https://www.qiagen.com). DNA concentration was determined using the NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific, Freemont, CA, https://www.thermofisher.com). Genetic analysis of gene was performed using a commercially available status kit (certified CE-IVD for diagnostic use; Diatech Pharmacogenetics, Jesi, Ancona, Italy, http://www.diatechpharmacogenetics.com/en/). The kit permits the identification of mutations in KRN 633 codons 542, 545, and 546 of exon 9 (E542K, E545K, E545A, E545G, Q546E, Q546K) and codons 1043, 1047, and 1049 of exon 20 (M1043I, H1047Y, H1047R, H1047L, G1049R, G1049S) of the gene. Real-time polymerase chain reaction (RotorGene 6000; Qiagen) was carried out using 30-ng DNA as template. Specific mutations were subsequently identified by pyrosequencing on a PyroMark Q96 ID (Qiagen). Statistical Analysis pCR was defined as the absence of invasive breast cancer in both the breast and the axilla. The association between baseline biomarkers.
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