Endothelin, Non-Selective

General, the pCR rates were similar in wild-type and =

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, DNA concentration was determined using the NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific, Freemont, CA, Genetic analysis of gene was performed using a commercially available status kit (certified CE-IVD for diagnostic use; Diatech Pharmacogenetics, Jesi, Ancona, Italy, 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.