Comparison of an automated cartridge-based system for mRNA assessment with central immunohistochemistry in the neoadjuvant GeparX trial.

2019 ◽  
Vol 37 (15_suppl) ◽  
pp. 3075-3075
Author(s):  
Carsten Denkert ◽  
Theresa Link ◽  
Paul Jank ◽  
Marianne Just ◽  
Claus Hanusch ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Clinical Trial ◽  
Mrna Expression ◽  
Expression Analysis ◽  
Hormone Receptors ◽  
Therapy Response ◽  
Her2 Positive ◽  
Ki 67 ◽  
Mrna Expression Analysis

3075 Background: Hormone receptors, HER2 and Ki-67 are prognostic values typically determined for breast cancer (BC) outcome and prediction of therapy response. A RT-qPCR based system, the Xpert Breast Cancer STRAT4, can be used to classify BC tissues regarding their hormone receptor status, HER2 and proliferation via Ki-67. We compared mRNA expression analysis of ER, PR, HER2, and Ki-67 by this automated in-vitro diagnostic platform (GeneXpert) (GX) with central immunohistochemistry (IHC) in a large clinical trial cohort. Methods: BC patients from the prospective GBG neoadjuvant trial GeparX (NCT02682693) (still recruiting) were included in this biomarker project. We used formalin-fixed paraffin embedded (FFPE) pretherapeutical core biopsies with a tumor content > 10%. One 4 µm FFPE tissue section was first processed with the Xpert FFPE Lysis Kit, the sample lysate was placed in the STRAT4 cartridge system and then tested on the GX system in which the purification, amplification and real-time detection took place within two hours automatically. Results: A total of 503 (99%) of the 509 samples had a valid measurement of all four genes. 258 samples (51.3%) of the cohort were classified in central pathology as ER positive, 196 (39%) as PR positive and 78 (15.5%) as HER2-positive, and 421 samples (83.7%) were Ki-67-high ( > 20%). The simple kappa coefficient was for ER = 0.7938, PR = 0.6540, HER2 = 0.8172 and Ki-67 = 0.3655. This indicates, that the measurements for ER, PR and HER2 showed a high correlation between both methods, whereas the measurement of Ki-67 does not. The accuracy between the STRAT4 and IHC was 89.7% for ER, 83.3% for PR, 94.6% for HER2 and 86.7% for Ki-67. According to molecular subgroups, highest accuracy regarding Ki-67, was determined in TNBC (96.2%; luminal: 81.1%; HER2-positive: 76.9%). Conclusions: Our results show a high concordance between standardized central IHC and automated mRNA expression analysis for the most important BC biomarkers ER, PR and HER2. For the proliferation marker Ki-67, the concordance is slightly lower. The STRAT4 assay might offer additional option to conventional methods for BC biomarker assessment, in particular in settings where IHC is not feasible. To determine the clinical validity, additional outcome analyses are necessary. Clinical trial information: NCT02682693.

scholarly journals Determination of Hormone Receptors, Human Epidermal Growth Factor Receptor 2 and Ki67 Status in Invasive Breast Carcinoma: A Concordance Study between Immunohistochemistry, Fluorescence in Situ Hybridization, and GeneXpert® Breast Cancer STRAT4 Assay

2021 ◽  
Author(s):  
rajaa elaje ◽  
Abdellah Naya ◽  
Ayoub KHOAJA ◽  
Younes Zaid ◽  
Mounia Oudghiri ◽  
...  
Keyword(s):  
Breast Cancer ◽  
In Situ Hybridization ◽  
Mrna Expression ◽  
Hormone Receptors ◽  
Therapy Response ◽  
Cancer Prognosis ◽  
University Hospital ◽  
Automated System ◽  
Ki 67

Abstract BackgroundThe accurate assessment of hormone receptors (ER and PR), HER2 and Ki-67 proliferative index provides meaningful information about breast cancer prognosis and prediction of therapy response. Immunohistochemistry, the most common method for evaluating these prognostic biomarkers, can be impacted by numerous variabilities due to pre-analytical/analytical factors and subjective interpretation by pathologists. The Xpert® Breast Cancer STRAT4, a RT-qPCR based system, can be used to classify breast invasive carcinomas based on the assessment of these 4 biomarkers. In this study, we investigated the accuracy of RT-qPCR based mRNA expression levels in a closed, single-use cartridge, automated system compared with the current gold standard, immunohistochemistry (IHC), and fluorescent in situ hybridization (FISH) for HER2 equivocal cases.Methods We evaluated ESR1, PGR, ERBB2 and MKi67 mRNA expression by Xpert Breast Cancer STRAT4 and ER, PR, HER2 and Ki67 by IHC (FISH for HER2 IHC 2+) in 200 formalin-fixed paraffin-embedded (FFPE) tissue blocks with invasive breast cancer, collected from the Pathology Department of Casablanca Ibn Rochd University Hospital.Results Concordance between Xpert ® Breast Cancer STRAT4 and IHC was 93.5% for ER, 83.51% for PR, 95% for HER2 (92% for IHC+FISH), and 81.20% for Ki67 (excluding intermediate IHC Staining 10 ≤ %IHC <20). The simple Kappa coefficient was, for ER, 0.830 (P < 0, 0001), 0.565 (P < 0, 0001) for PR, 0.838 (P < 0, 0001) for HER2-IHC, 0.771 (P< 0, 0001) for HER2 IHC+FISH and, for, Ki67, 0.458 (P < 0, 0001).Conclusions We demonstrated globally a high concordance between centrally assessed IHC, IHC+FISH and mRNA measurements of ER/ESR1 and HER2/ERBB2, and a moderate agreement between PR/PGR and Ki67/MKi67. These findings provide an additional, objective, and quantitative assessment of tumor receptor status in breast cancer.

scholarly journals Novel Molecular Markers for Breast Cancer

2016 ◽  
Vol 8 ◽  
pp. BIC.S38394 ◽  
Author(s):  
Kazushi Inoue ◽  
Elizabeth A. Fry
Keyword(s):  
Breast Cancer ◽  
Molecular Markers ◽  
Hormone Receptors ◽  
Epithelial Mesenchymal Transition ◽  
Disease Free Survival ◽  
Her2 Positive ◽  
Specific Therapy ◽  
Predictive Values ◽  
Mesenchymal Transition

The use of molecular biomarkers assures that breast cancer (BC) patients receive optimal treatment. Established biomarkers, such as estrogen receptor, progesterone receptor, HER2, and Ki67, have been playing significant roles in the subcategorization of BC to predict the prognosis and decide the specific therapy to each patient. Antihormonal therapy using 4-hydroxytamoxifen or aromatase inhibitors have been employed in patients whose tumor cells express hormone receptors, while monoclonal antibody to HER2 has been administered to HER2-positive BCs. Although new therapeutic agents have been developed in the past few decades, many patients still die of the disease due to relapse; thus, novel molecular markers that predict therapeutic failure and those that can be targets for specific therapy are expected. We have chosen four of such molecules by reviewing recent publications, which are cyclin E, B-Myb, Twist, and DMP1β. The oncogenicity of these molecules has been demonstrated in vivo and/or in vitro through studies using transgenic mice or siRNAs, and their expressions have been shown to be associated with shortened overall or disease-free survival of BC patients. The former three molecules have been shown to accelerate epithelial-mesenchymal transition that is often associated with cancer stem cell-ness and metastasis; all these four can be novel therapeutic targets as well. Thus, large prospective studies employing immunohistochemistry will be needed to establish the predictive values of these molecules in patients with BC.

scholarly journals Neoadjuvant Chemotherapy Exerts Selection Pressure Towards Luminal Phenotype Breast Cancer

Breast Care ◽  
2017 ◽  
Vol 12 (6) ◽  
pp. 391-394 ◽  
Author(s):  
Giulia Galli ◽  
Giacomo Bregni ◽  
Stefano Cavalieri ◽  
Luca Porcu ◽  
Paolo Baili ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Neoadjuvant Chemotherapy ◽  
Hormone Receptors ◽  
Residual Disease ◽  
Luminal A ◽  
Her2 Positive ◽  
Ki 67 ◽  
Surgical Specimen ◽  
Luminal B ◽  
Basal Phenotype

Background: Breast cancer (BC) phenotype after neoadjuvant chemotherapy (NAC) has not been extensively described and few data exist on whether expression of the primary tumor hormone receptors, HER2 and Ki-67 changes as a result of chemotherapy. Materials and Methods: We analyzed specimens from all BC patients treated with anthracycline/taxane-based NAC at our Institution between January 2010 and March 2015 (n = 325). The expression of estrogen receptor (ER), progesterone receptor (PR), HER2 and Ki-67 was determined in pre- and post-NAC specimens. McNemar's test was used to compare paired proportions. Results: Among patients with residual disease after NAC, basal phenotype was luminal A, luminal B, HER2 positive and triple negative in 44, 111, 74 and 27 cases, respectively. PR-positive tumors decreased from 68.0% in the initial biopsy sample to 61.7% in the surgical specimen (p = 0.024). A Ki-67 of < 20% increased from 23.6% to 45% (p < 0.001). ER expression changed from positive to negative in 5% and from negative to positive in 16.7% of cases. Overall, 30% of cases underwent subtype changes, 79% of them towards luminal differentiation. Conclusions: The switch towards luminal phenotype suggests some kind of endocrine effect of NAC. Our findings raise renewed interest in combinatorial cytotoxic chemotherapy with concomitant or rather sequential endocrine therapy, either alone or with targeted agents.

scholarly journals Development of a Ki-67-based clinical trial assay for neoadjuvant endocrine therapy response monitoring in breast cancer

2017 ◽  
Vol 165 (2) ◽  
pp. 355-364 ◽  
Author(s):  
Rodrigo Goncalves ◽  
Katherine DeSchryver ◽  
Cynthia Ma ◽  
Yu Tao ◽  
Jeremy Hoog ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Clinical Trial ◽  
Endocrine Therapy ◽  
Therapy Response ◽  
Response Monitoring ◽  
Neoadjuvant Endocrine Therapy ◽  
Ki 67

AGE FEATURES OF MOLECULAR-BIOLOGICAL SUBTYPES OF BREAST CANCER

2020 ◽  
Vol 17 (2) ◽  
pp. 187-192
Author(s):  
E.A. Novikova ◽  
◽  
O.V. Kostromina ◽  
D.V. Mikhailov ◽  
S.L. Leontiev ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Hormone Receptors ◽  
Age Groups ◽  
Luminal A ◽  
Ki 67 ◽  
Luminal B ◽  
Age Related ◽  
Age Features ◽  
Immunohistochemical Studies ◽  
Triple Negative Subtype

Aim. The aim of the study was to determine the presence of peculiarities of the age structure in patients with various surrogate molecular biological subtypes of breast cancer. Materials and research methods. This work analyzes the age-related characteristics of the occurrence of molecular biological subtypes in 499 patients with invasive breast cancer. All cases were divided into 5 molecular biological subtypes based on immunohistochemical studies of hormone receptors, Her2, Ki-67. The average age of the patients was 53.4±0.39 years, the predominant group was patients from 50 to 60 years (37.2% of the total). Research results. In patients under 40 years old, the triple negative subtype prevailed (44.8%). Luminal A subtype prevailed in the groups 51-60 years old (more than 41.4%) and over 60 years old (39.7%). Luminal B (Her2-) subtype was equally found in all age groups.

scholarly journals Splicing factor SRSF1 promotes breast cancer progression via oncogenic splice switching of PTPMT1

2021 ◽  
Vol 40 (1) ◽  
Author(s):  
Jun-Xian Du ◽  
Yi-Hong Luo ◽  
Si-Jia Zhang ◽  
Biao Wang ◽  
Cong Chen ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Progression ◽  
High Risk Group ◽  
Clinical Samples ◽  
Binding Motif ◽  
Ki 67 ◽  
Tissue Samples

Abstract Background Intensive evidence has highlighted the effect of aberrant alternative splicing (AS) events on cancer progression when triggered by dysregulation of the SR protein family. Nonetheless, the underlying mechanism in breast cancer (BRCA) remains elusive. Here we sought to explore the molecular function of SRSF1 and identify the key AS events regulated by SRSF1 in BRCA. Methods We conducted a comprehensive analysis of the expression and clinical correlation of SRSF1 in BRCA based on the TCGA dataset, Metabric database and clinical tissue samples. Functional analysis of SRSF1 in BRCA was conducted in vitro and in vivo. SRSF1-mediated AS events and their binding motifs were identified by RNA-seq, RNA immunoprecipitation-PCR (RIP-PCR) and in vivo crosslinking followed by immunoprecipitation (CLIP), which was further validated by the minigene reporter assay. PTPMT1 exon 3 (E3) AS was identified to partially mediate the oncogenic role of SRSF1 by the P-AKT/C-MYC axis. Finally, the expression and clinical significance of these AS events were validated in clinical samples and using the TCGA database. Results SRSF1 expression was consistently upregulated in BRCA samples, positively associated with tumor grade and the Ki-67 index, and correlated with poor prognosis in a hormone receptor-positive (HR+) cohort, which facilitated proliferation, cell migration and inhibited apoptosis in vitro and in vivo. We identified SRSF1-mediated AS events and discovered the SRSF1 binding motif in the regulation of splice switching of PTPMT1. Furthermore, PTPMT1 splice switching was regulated by SRSF1 by binding directly to its motif in E3 which partially mediated the oncogenic role of SRSF1 by the AKT/C-MYC axis. Additionally, PTPMT1 splice switching was validated in tissue samples of BRCA patients and using the TCGA database. The high-risk group, identified by AS of PTPMT1 and expression of SRSF1, possessed poorer prognosis in the stage I/II TCGA BRCA cohort. Conclusions SRSF1 exerts oncogenic roles in BRCA partially by regulating the AS of PTPMT1, which could be a therapeutic target candidate in BRCA and a prognostic factor in HR+ BRCA patient.

scholarly journals O80: GREMLIN-1 PROMOTES TUMOURIGENESIS AND METASTASIS IN HER2 POSITIVE BREAST CANCER

2021 ◽  
Vol 108 (Supplement_1) ◽  
Author(s):  
C Zabkiewicz ◽  
L Ye ◽  
R Hargest
Keyword(s):  
Breast Cancer ◽  
Cellular Growth ◽  
Breast Cancers ◽  
Her2 Positive ◽  
Mesenchymal Transition ◽  
Her2 Breast Cancer ◽  
Bmp Signalling ◽  
Gremlin 1

Abstract Introduction HER2 over-expression denotes poor prognosis in breast cancers.Bone morphogenetic protein(BMP) signalling is known to interact with EGF signalling, co-regulating breast cancer progression.BMP antagonist Gremlin-1 may influence breast cancer disease progression, but this remains unexplored in HER2 positive breast cancers. Method GREM1 and HER2 expression, and clinical outcomes were examined in clinical cohorts.GREM1 overexpression or pEF control plasmid were transduced into BT474 HER2+breast cancer cells. In vitro function tests using BT474 pEF and BT474GREM1cells include 2D/3D growth, migration, and expression of epithelial to mesenchymal transition(EMT)markers. Signalling cascades were examined in BT474 treated with RhGremlin-1. In vivo, BALB/c nude mice underwent either mammary injection or intra-cardiac injection of BT474pEF or BT474GREM1 cells and disease burden assessed. Result GREM1 expression correlates with HER2 in breast tumours(p=0.03) and is higher in metastatic HER2 positive cancers (p = 0.04). HER2 positive patients with high GREM1 have poor survival(p = 0.0002). BT474GREM1cells have up-regulated markers of EMT compared to control. BT474 RhGremlin-1 treated cells have active AKT pathway signalling, independent of BMP signalling. In vitro,  BT474GREM1cells significantly proliferate and migrate compared to control(p&lt;0.05 and p &lt; 0.001).This is confirmed in vivo,  BT474GREM1 mice grew significantly larger mammary tumours(p&lt;0.05) and had more PETCT metastatic hotspots. Conclusion Gremlin-1 is correlated with poor outcomes in HER2 patients and promotes breast cancer cellular growth, migration and metastasis.Gremlin-1 is a novel area of research with potential as a prognostic biomarker and therapeutic target for personalised, effective, breast cancer outcomes. Take-home message BMP antagonists are gaining interest for their potential in breast cancer prognosis and therapeutics.This novel area of research shows BMP antagonist Gremlin-1 is of importance in HER2 positive breast cancers. DRAGONS DEN

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