EndoPredict and Mammaprint Risk Classification

EndoPredict and Mammaprint Risk Classification equation of jeopardize classification between EndoPredict and Mammaprint in ER- overbearing/HER2-negative primary invasive chest of drawers cancer.Alberto Pelez-Garcia, Laura Yebenes, Alberto Berjon, Antonia Angulo, Pilar Zamora, Jose Ignacio Snchez-Mendez, Enrique Espinosa, Andres Redondo, Victoria Heredia, Marta Mendiola, Jaime Feliu, David HardissonCorresponding Author David Hardisson, MD, PhD De circumstancesment of Pathology infirmary Universitario La Paz, IdiPAZ Paseo de la Castellana, 261 28046 Madrid, Spain.ABSTRACTPurposeTo compare the prognostic mathematical operation of the EndoPredict assay with the MammaPrint chumps obtained for the same cancer samples on 40 estrogen-receptor positive/HER2-negative depreciator carcinomas.MethodsFormalin-fixed, paraffin-embedded invasive dummy carcinoma create from raw materials that were previously study with MammaPrint as part of good turn care of the patients, andwere classified as high-risk (20 patients) and low-risk (20 patients), were selected to be analyzed by the EndoPredict assay, a due south ingredientration gene saying shew that combines looking of 8 genes (EP grade) with two clinicopathological factors (neoplasm size and nodal status, EPclin score).ResultsThe EP score classified 15 patients as low-risk and 25 patients as high-risk. EPclin re-classified 5 of the 25 EP high-risk patients into low-risk, resulting in a total of 20 high-risk and 20 low-risk tumours. EP score and MammaPrint score were significantly correlated (p=0.008). Twelve of 20 samples classified as low-risk by MammaPrint were as well as low-risk by EP score (60%). 17 of 20 MammaPrint high-risk tumors were also high-risk by EP score. The over any capital of New Hampshire between EP score and MammaPrint was 72.5%. EPclin score also correlated with MammaPrint results (p=0.004). Discrepancies between both leavens occurred in 10 cases 5 MammaPrint low-risk patients were classi fied as EPclin high-risk and 5 high-risk MammaPrint were classified as low-risk by EPClin (overall concordance 75%).ConclusionsThis study demonstrates a moderate concordance between MammaPrint and EndoPredict. Differences in results could be explained by the inclusion of different gene sets in each platform, and the inclusion of clinical parameters, such(prenominal) as tumor size and nodal status, in the EndoPredict test.KeywordsBreast cancer prognosis gene expression signatures EndoPredict MammaPrintINTRODUCTIONBreast cancer is the most common cancer and the second most frequent cause of cancer death among women in real countries. Approximately 231,840 new cases of invasive rapper cancer and 40,290 deaths are expect among US women in 2015 1.Currently, the decision on adjuvant treatment for breast cancer patients is based on risk assessment exploitation clinicopathological criteria, such as patient age, menopausal status, axillary lymph node status, tumor size, tumor grade, estro gen receptor (ER)/progesterone receptor (PgR) expression, HER2 status, and Ki67 score. However, decision making in adjuvant treatment of women with ER-positive/HER2-negative early breast cancer remains a difficult task. Routinely, all of these patients will receive adjuvant hormonal treatment. However, a substantial proportion of these patients are also treated with adjuvant chemotherapy, although a significant part of these will not achieve a pass on reduction of their risk of return 2.Therefore, a major challenge for clinical oncologists is to identify those patients who will not benefit for adjuvant chemotherapy, and those who are more than likely to develop recurrence, so that the most appropriate therapeutic regime can be administered 2, 3.In recent years, molecular characterization of breast cancer has contributed to broaden our understanding of breast cancer as a heterogeneous disease, and led to the development of a variety of prognostic and prophetic gene signatures 4. Morever, these assays may also be useful in recurrence prediction and treatment decision making 5. One of the most widely employ tests is the MammaPrint (MP) assay (Agendia Laboratories, Amsterdam, The Netherlands), which is a prognostic score performed by a central laboratory that was cleared by the FDA in 2007. MP was ab initio limited by its requirement for fresh tissue, but it is now pass for formalin-fixed, paraffin-embedded (FFPE) tissue 6. MP measures the expression of 70 genes using a microarray platform, and reports a binary risk classification (low-risk or high-risk) for recurrence without adjuvant chemotherapy. This tuition is intended to spare patients at low-risk of recurrence from receiving adjuvant chemotherapy, with its attendant morbidity. It is not intended to predict the response, per se, to chemotherapy rather, it helps to select patients who are likely to benefit from chemotherapy from a prognostic point of view 7. More recently developed, the EndoPredict assay (EP) (Sividon nosology GmbH, Cologne, Germany), is a diagnostic test based on gene expression data in combination with clinicopathological risk parameters to assess the risk of outside metastasis in patients with ER-positive/HER2-negative primary breast cancer if treated with adjuvant ductless gland therapy alone 8. This test measures the expression of eight cancer-related genes of interest (BIRC5, UBE2C, DHCR7, RBBP8, IL6ST, AZGP1, MGP and STC2) and common chord credit entry genes (CALM2, OAZ1 and RPL37A) to calculate a molecular risk score (EP score). The molecular risk score is then combined with the nodal status and tumor size resulting in a molecular-clinicopathological hybrid score (EPclin score) with improved prognostic power. Using a predefined cutoff value, patients are stratified into low- or high-risk of distant recurrence. The test can be carried out on identification numberly processed and archived FFPE tissue, and is intentional to be performed decentrally 9, 10. EP was validated in three randomized endocrine pattern III trials with patients with ER-positive/HER2-negative node negative and node positive breast carcinomas 5, 8. The EP provided surplus prognostic information to conventional risk factors such as grading, valued ER, or Ki67 and outperformed risk classification by clinical guidelines. Moreover, it could be exhibit that EP is prognostic for early and late metastasis 5, 11.The EPclin score was also directly compared to stringently clinical risk classifications (like St. Gallen, German S3, and NCCN) and found to be superior to these classifiers 11.The accusive of this study was to compare the concordance of EndoPredict results in 40 ER-positive/HER2-negative breast carcinomas which were previously tested with MammaPrint and categorized as low-risk (20 patients) or high-risk (20 patients). We further judge TargetPrint (Agendia Laboratories), a commercially available mribonucleic acid-based gene expression test that qua ntitatively determines gene expression levels of ER, PgR, and HER2.MATERIALS AND METHODSPatients and tumor samplesThis study involved 40 patients with ER-positive/HER2-negative early-stage breast carcinoma. All patients underwent surgery between March 2012 and December 2015 at the University Hospital La Paz, Madrid, Spain. Data on age and tumor characteristics were collected for all patients. The surgical specimens were fixed in 10% buffered formalin and embedded in paraffin. Four-m thick sections were stained with hematoxylin-eosin for histologic diagnosis. Sections (10m) with at least 40% of tumor cellularity were selected for the study.Immunohistochemistry for ER/PR/HER2 and Ki67 and Fluorescence in situ Hybridization (FISH) for HER2All cases were reviewed by two breast pathologist (DH and LY) to assess tumor grade (using the Nottingham histological three-tier grading system), tumor size, nodal status, ER, PgR, HER-2, and Ki67 expression. The expression of ER (clone EP1 Dako, Gl ostrup, Denmark, prediluted), PgR (clone PgR1294 Dako, prediluted), and Ki67 (clone MIB1 Dako, prediluted) were determined by immunohistochemistry (IHC) during routine pathologic examination. ER and PgR status was determined based on the role of positive nuclei in the invasive neoplastic compartment of the tissue. Tumors were classified as ER- or PgR-positive when 1% invasive tumor cells showed definite nuclear dapple, regardless of maculation intensity. Ki67 was evaluated as the percentage of positively stained nuclear cancer cells (regardless of staining intensity). HER2 expression was evaluated with the HercepTest kit out (Dako) and scored as 0, 1+, 2+, or 3+, gibe to the FDA scoring system. Tumors scored as 2+ were re-tested with FISH using the HER2 IQFISH PharmDx kit (Dako).Mammaprint TestThe MammaPrint test was performed on representative paraffin blocks at the centralise Agendia Laboratories (Amsterdam, The Netherlands) blinded for clinical and histological data as part of routine care of the patients included in this study. Additionally to MammaPrint, TargetPrint assay, an additional test that is an preference measurement of ER, PgR, and HER2 to IHC/FISH assessment, was also performed.EndoPredict TestThe same tumor tissue block used for MammaPrint testing in each case was used for EP test. RNA extraction was performed as previously described 9. Total RNA was extracted from one 10-m whole formalin-fixed, paraffin-embedded tissue section using a silica-coated magnetic bead-based method with Tissue Preparation Reagents (Sividon Diagnostics). Expression of eight genes-of-interest (AZGP1, BIRC5, DHCR7, IL6ST, MGP, RBBP8, STC2, UBE2C), three normalization genes (CALM2, OAZ1, RPL37A) as well as the amount of residual genomic desoxyribonucleic acid (HBB) were assessed by the EP assay (Sividon Diagnostics). Gene expression was assessed by one-step RT-qPCR using the SuperScript III PLATINUM One-Step Quantitative RT-PCR System with ROX (Invitrogen, Karlsru he, Germany) according to manufacturers instructions in a VERSANT kPCR Molecular System (Siemens health care Diagnostics, Erlangen, Germany). EP and EPclin scores were determined as published earlier 8, 9 using the EndoPredict bailiwick Generator software which is available online (www1.endopredict.com). The predefined cut-offs for diagnostic decisions were applied to stratify patients into low- or high-risk groups EP low-risk (