Human Epidermal Growth Factor Receptor-2 Testing Research Articles

Human epidermal growth factor receptor-2 expression and subsequent dynamic changes in patients with ovarian cancer

Human epidermal growth factor receptor-2 (HER2)-targeting drugs are increasingly being incorporated into therapeutic paradigms for non-breast cancers, yet studies on HER2 expression in ovarian cancer (OC) are inadequate. Here, we studied the HER2 status and dynamic changes in OC by reviewing the records of patients who underwent HER2 testing at a single institution. Clinical parameters, including histology, BRCA status, and immunohistochemistry (IHC), were evaluated alongside HER2 expression, timing, and anatomical location. Among 200 patients, 28% and 6% exhibited expression scores of 2+ and 3+, respectively. HER2 3+ scores were observed in 23%, 11%, 9%, and 5% of mucinous, endometrioid, clear cell, and high-grade serous tumors, respectively, and were exclusively identified in BRCA-wildtype, mismatch repair-proficient, or PD-L1-low-expressing tumors. The TP53 mutation rate was low, whereas ARID1A, KRAS, and PIK3CA mutations were relatively more prevalent with HER2 scores of 2+ or 3+ than with 0 or 1+. Four of the five tumors with an HER2 3+ score exhibited ERBB2 amplification. Among 19 patients who underwent multiple time-lagged biopsies, 11 showed increased HER2 expression in subsequent biopsies. Patients with HER2-overexpressing OC exhibited distinct histological, IHC, and genomic profiles. HER2-targeting agents are potential options for BRCA-wildtype patients, particularly as later lines of treatment.

Expression of HER2 in high-grade urothelial carcinoma based on Chinese expert consensus and the clinical effects of disitamab vedotin-tislelizumab combination therapy in the treatment of advanced patients.

Background: A vast number of researchers have discovered high levels of human epidermal growth factor receptor-2 (HER2) expression in urothelial carcinoma (UC), but they do not use a uniform scoring system. Based on the 2021 edition of clinical pathological expert consensus on HER-2 testing in UC in China, we investigated the expression level and clinical significance of HER2 in high-grade UC. Furthermore, we looked at the prognosis of patients with locally advanced/metastatic UC after combining HER2 targeting antibody-drug conjugates (ADC) medication disitamab vedotin (DV) with programmed cell death protein 1 (PD-1) inhibitor tislelizumab. Patients and methods: From 2019 to 2022, we collected paraffin specimens of UC from the Department of Urology at the Provincial Hospital Affiliated to Shandong First Medical University. HER2 expression-related factors were investigated. Patients with advanced UC who have failed systemic chemotherapy at least once and had received immune checkpoint inhibitor (ICI) medication during second-line treatment were selected and treated with DV in combination with tislelizumab. We assessed the therapy's efficacy and safety. Results: 185 patients with high-grade UC were included in this investigation. 127 patients (68.7%) were HER2 positive (IHC 2+/3+) according to the 2021 Clinical pathological expert consensus on HER2 testing in UC in China. The clinical stage of UC differed statistically significantly between the HER2-and HER2+ groups (p = 0.019). Sixteen advanced UC patients were treated with DV and tislelizumab for a median of 14months. The disease control rate was 87.5%, while the objective response rate (ORR) was 62.5%. The ORR of HER2+ individuals was higher than that of HER2-individuals (70.0% vs. 50.0%). The median progression-free survival or overall survival was not reached. In this study, the incidence of treatment-related adverse events was 68.8% (11/16), with all of them being grade 1 or 2 adverse reactions. Conclusion: HER2 protein expressed at a high percentage in UC, and 68.7% patients expressed HER2 positive (IHC 2+/3+). HER2+ expression is positively correlated with higher clinical stage of UC. HER2 targeted ADC drug disitamab vedotin combining with PD-1 inhibitor tislelizumab has shown efficacy, safety and controllable adverse reactions in the treatment of advanced UC.

Diagnostics and treatment for HER2+ mG&CRC: Learning outcomes of GetSMART.

82 Background: Gastric/colorectal cancer (G&CRC) are among the most prevalent cancers. Efforts are ongoing to better assess their clinical/genomic factors. HER2 (human epidermal growth factor receptor 2) testing is strongly recommended in clinical practice to guide treatment. An accredited education program (GetSMART) was launched online (January 2021–April 2022) to equip healthcare providers (HCPs) with knowledge, skills, and confidence to accurately assess/apply molecular testing and therapy options for metastatic G&CRC patients. Methods: A knowledge/confidence-based assessment was embedded in four modules to measure learners’ knowledge mastery (% answering correctly with confidence) pre/post-activity. Self-reported confidence and anticipated performance in clinical practice were measured before/after each module. At the end of program, HCPs were asked about their commitment to change. McNemar’s test was applied to assess statistical significance in pre-post for paired items, and descriptive analysis for non-paired items. Results: Overall, 925 unique HCPs engaged in GetSMART, 31% (n=284) completed all modules, of which 57% (n=163) consented analysis of their data. A statistically significant increase (p<0.05) was measured in percentage of HCPs demonstrating knowledge mastery, confidence, and commitment to change their practice from pre to post across four competencies, except for percentage of HCPs “often”/“always” engaging patients in shared decision-making (SDM) (3, Performance). Largest gains in knowledge mastery/confidence were noted for HCPs’ ability to anticipate/manage adverse events (AE) ( 4A); and for anticipated performance, in HCPs considering appropriate treatment. Post-activity, 66% (98/149) of HCPs intended to change practice based on module content, 83% (70/84) of whom were “highly”/“somewhat” confident to make changes. The activity’s anticipated impact on patients was rated “major”/“moderate” by 69% (103/149) of HCPs. Conclusions: Identification of HER2 aberrations in mG&CRC and its appropriate treatment selection remains challenging in oncology care. This evaluation demonstrated that GetSMART positively impacted HCPs’ knowledge, confidence, and anticipated performance in clinical practice to enhance health outcomes of HER2+ mG&CRC patients. [Table: see text]

Comparison of breast cancer HER-2 receptor testing with immunohistochemistry and in situ hybridization.

Human epidermal growth factor receptor-2 (HER2) status can be tested with immunohistochemistry (IHC) and in situ hybridization (ISH). The 2018 American Society of Clinical Oncology/College of American Pathologists (ASCO/CAP) HER2 testing guidelines suggest initial HER2 testing using IHC and further testing IHC equivocal cases with ISH. However, many institutions perform both IHC and ISH on the same specimen. This study aims to analyze the concordance between HER2 IHC and ISH in order to evaluate the benefit of repeating HER2 testing on the same breast cancer specimens. Patients diagnosed with invasive breast cancer through BreastScreen NSW Sydney West program between January 2018 and December 2020 were identified and their HER2 IHC and HER2 ISH results on core needle biopsy (CNB) and surgical excisions (SE) were retrospectively collected. Specimens with both IHC and ISH results were then analyzed for agreement and concordance using unweighted kappa values. Equivocal IHC (2+) cases were excluded from concordance analysis. Overall, there were 240 invasive breast cancer specimens (CNB and SE) with both IHC and ISH recorded. Concordance between HER2 IHC and ISH was 100% (95% CI: 96.2-100%; κ=1.00 (P<0.001)). Of the IHC equivocal cases (n=146), 94.5% were ISH negative. There was perfect positive concordance and agreement between non-equivocal IHC and ISH results. This reinforces that IHC alone can be utilized reliably for testing HER2 status of non-equivocal cases consistent with the 2018 ASCO/CAP guidelines.

155P Artificial Intelligence (AI) - powered human epidermal growth factor receptor-2 (HER2) and tumor-infiltrating lymphocytes (TIL) analysis for HER2-positive early breast cancer patients treated with HER2-targeted neoadjuvant chemotherapy (NAC)

HER2 is an important predictive biomarker of HER2-targeted therapies for breast cancer. Reliable HER2 testing is essential for clinical practice, but immunohistochemistry (IHC) staining results are susceptible to considerable interobserver variations.

Variability in Breast Cancer Biomarker Assessment and the Effect on Oncological Treatment Decisions: A Nationwide 5-Year Population-Based Study.

Simple SummaryBiomarkers that define breast cancer treatment recommendations include estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth-factor receptor 2 (HER2); histological grade; and in many countries, the Ki67 proliferation index. However, the subjective nature and degree of variability in breast cancer biomarker assessment might result in under- or overtreatment. We demonstrated that limited variability exists in ER, PR, and HER2 positivity rates among 29 departments in Sweden, including 43,261 patients. However, even a few outlier labs affect endocrine and anti-HER2 treatment rates in a clinically relevant proportion, indicating a need for improvement. Despite international guidelines, standardized protocols, and external quality control procedures, very high variability was found in Ki67 scoring and histological grading, indicating a need for new methods. Monitoring rates of biomarker expression and treatments among departments should be mandatory in order to detect variability issues affecting the clinical management of breast cancer.We compared estrogen receptor (ER), progesterone receptor (PR), human epidermal growth-factor receptor 2 (HER2), Ki67, and grade scores among the pathology departments in Sweden. We investigated how ER and HER2 positivity rates affect the distribution of endocrine and HER2-targeted treatments among oncology departments. All breast cancer patients diagnosed between 2013 and 2018 in Sweden were identified in the National Quality Register for Breast Cancer. Cases with data on ER, PR, HER2, Ki67, grade, and treatment were selected (43,261 cases from 29 departments following the guidelines for biomarker testing). The ER positivity rates ranged from 84.2% to 97.6% with 6/29 labs out of the overall confidence intervals (CIs), while PR rates varied between 64.8% and 86.6% with 7/29 labs out of the CIs. HER2 positivity rates ranged from 9.4% to 16.3%, with 3/29 labs out of the overall CIs. Median Ki67 varied between 15% and 30%, where 19/29 labs showed significant intra-laboratory variability. The proportion of grade-II cases varied between 42.9% and 57.1%, and 13/29 labs were outside of the CI. Adjusting for patient characteristics, the proportion of endocrine and anti-HER2 treatments followed the rate of ER and HER2 positivity, illustrating the clinical effect of inter- and intra-laboratory variability. There was limited variability among departments in ER, PR, and HER2 testing. However, even a few outlier pathology labs affected endocrine and HER2-targeted treatment rates in a clinically relevant proportion, suggesting the need for improvement. High variability was found in grading and Ki67 assessment, illustrating the need for the adoption of new technologies in practice.

Receptor-Defined Breast Cancer in Five East African Countries and Its Implications for Treatment: Systematic Review and Meta-Analysis.

PURPOSEEstrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor-2 (HER2) are determinants of treatment and mortality for patients with breast cancer (BC). In East Africa, the estimated 5-year survival (37.7%) is far lower than the US average (90%). This meta-analysis investigates BC receptor subtypes within five East African countries to ascertain cross-country patterns and prioritize treatment needs.METHODSFrom a PubMed search, January 1, 1998-June 30, 2019, for all English-only BC articles for Ethiopia, Kenya, Rwanda, Tanzania, and Uganda, eligible studies had receptor distributions for female BC samples ≥ 30 patients. Outcomes were proportions of ER+, PR+, and HER2-positive (HER2+), and/or molecular subtypes. Data included study characteristics and mean or median patient age. Using metaprop, Stata 16, we estimated pooled proportions (ES) with 95% CIs and assessed heterogeneity.RESULTSAmong 36 BC studies with receptor data, 21 met criteria. Weighted mean age was 47.5 years and median, 48. Overall ES were as follows: 55% for ER-positive (ER+) (95% CI, 47 to 62), 23% for HER2+ (95% CI, 20 to 26), and 27% for triple-negative BC (TNBC) (95% CI, 23 to 32).CONCLUSIONWe found differences between countries, for example, lower distribution of TNBC in Ethiopia (21%) compared with Uganda (35%). ER+, the dominant BC subtype overall at 55%, emphasizes the need to prioritize endocrine therapy. Overall proportions of HER2+ BC (with or without ER+ or PR+), 23%, approached proportions of TNBC, 27%, yet HER2 testing and treatment were infrequent. Testing and reporting of receptor subtypes would promote delivery of more effective treatment reducing the mortality disparity.

Predictors of pathological complete response to neoadjuvant treatment and changes to post-neoadjuvant HER2 status in HER2-positive invasive breast cancer

The response of human epidermal growth factor receptor2 (HER2)- positive breast cancer (BC) patients to anti-HER2 targeted therapy is significant. However, the response is not uniform and a proportion of HER2-positive patients do not respond. This study aims to identify predictors of response in the neoadjuvant treatment and to assess the discordance rate of HER2 status between pre- and post-treatment specimens in HER2-positive BC patients. The study group comprised 500 BC patients treated with neoadjuvant chemotherapy (NACT) and/or neoadjuvant anti-HER2 therapy and surgery who had tumours that were 3+ or 2+ with HER2 immunohistochemistry (IHC). HER2 IHC 2+ tumours were classified into five groups by fluorescence in situ hybridisation (FISH) according to the 2018 ASCO/CAP guidelines of which Groups 1, 2 and 3 were considered HER2 amplified. Pathological complete response (pCR) was more frequent in HER2 IHC 3+ tumours than in HER2 IHC 2+/HER2 amplified tumours, when either in receipt of NACT alone (38% versus 13%; p = 0.22) or neoadjuvant anti-HER2 therapy (52% versus 20%; p < 0.001). Multivariate logistic regression analysis showed that HER2 IHC 3+ and histological grade 3 were independent predictors of pCR following neoadjuvant anti-HER2 therapy. In the HER2 IHC 2+/HER2 amplified tumours or ASCO/CAP FISH Group 1 alone, ER-negativity was an independent predictor of pCR following NACT and/or neoadjuvant anti-HER2 therapy. In the current study, 22% of HER2-positive tumours became HER2-negative by IHC and FISH following neoadjuvant treatment, the majority (74%) HER2 IHC 2+/HER2 amplified tumours. Repeat HER2 testing after neoadjuvant treatment should therefore be considered.

The differences of clinicopathologic characteristics among subgroups of reclassified HER2 fluorescence in situ hybridization (FISH) according to the ASCO/CAP 2018 breast cancer HER2 testing guidelines

AimsThe aim of this study is to analyse differences in clinicopathologic features among reclassified human epidermal growth factor receptor-2 (HER2) fluorescence in situ hybridization (FISH) results in breast cancers according...

Abstract P1-03-02: ASCO/CAP human epidermal growth factor receptor-2 (HER2) in situ hybridization (ISH) categories evaluated by quantitative HER2 protein diagnostic methodologies: A comparative analysis

Abstract Background: In 2013, the ASCO/CAP consensus panel published updated guidelines for HER2 testing in breast cancer that modified the definition of HER2 amplification by in situ hybridization (ISH), creating five new prognostic categories (group 1: classic amplified, group 2: monosomy, group 3: co-amplified (polysomy), group 4: equivocal, and group 5: classic non-amplified). Patients determined to be ISH amplified, were considered eligible for HER2-directed therapy. Concern over whether patients from non-classic groups 2-4 would benefit from treatment has led to the recent publication of the 2018 HER2 focused update. This update has modified the criteria for interpreting these ISH categories, recommending that the final diagnosis take into consideration a combination of HER2 immunohistochemistry (IHC) and ISH results. With increased emphasis on the HER2 protein assessment, it has prompted us to quantitatively examine HER2 protein expression in the ISH categories, using two different novel technologies. Materials &amp; Methods: A cohort of 170 cases (URMC) and 102 cases (PSHMC) of invasive breast cancers, which had previously undergone HER2 IHC and ISH testing, were selected for this study. Cases were sorted and categorized into the HER2 ISH categories defined by ASCO/CAP. HER2 protein expression was quantitatively measured in the URMC and PSHMC cohorts using a novel immunodetection methodology (streptavidin-coated Phosphor-Integrated Dot (PID) fluorescent nanoparticles), and a novel dual-antibody, proximity-binding immunoassay (HERmark® Breast Cancer Assay, Monogram Biosciences, South San Francisco, California), respectively. HER2 protein expression was compared to the HER2 FISH and IHC results by ASCO/CAP category. Results: Cases in group 1 had a significantly (p &amp;lt; 0.01) higher average PID/cell and HERmark compared to cases in groups 2-5 (Table 1). Cases in groups 2-4 showed lower quantitative levels of HER2 protein expression, similar to the classic non-amplified cases (group 5). Group 1 was further divided into three subgroups (Table 2): Group A - ISH high-level amplified (ratio &amp;gt; 2, HER2 &amp;gt; 6, CEP17 &amp;lt; 2.7), Group B - amplified with elevated CEP17 (ratio &amp;gt; 2, CEP17 &amp;gt; 2.7), and Group C - low-level amplified (ratio &amp;gt; 2, HER2 &amp;gt; 4 and &amp;lt; 6). Group A and B had a significantly (p &amp;lt; 0.01) higher average PID/cell and HERmark compared to Group C. Group C was more comparable to cases in groups 2-5 (Table 1). Conclusion: Our results suggest that quantitative assessment of HER2 protein expression may help to further classify cases for HER2 status for targeted therapy, supporting the 2018 ASCO/CAP recommendation that non-classic ISH results might be resolved by evaluating protein expression. Follow up studies with a larger patient cohort and dual quantitative assessment are warranted. Average PID/cell and HERmark in ASCO category groupsASCO category groupN (URMC)PID/cell (URMC)*N (PSHMC)HERmark (PSHMC)*18888.07761.521011.20N/A32016.0213.84238.5315.95296.3208.3*averageTable 2:Average PID/cell and HERmark in subgroups of Group 1SubgroupN (URMC)PID/cell (URMC)*N (PSHMC)HERmark (PSHMC)*A24157.66465.7B34101.61044.1C3016.9329.8*average Citation Format: Buscaglia B, Turner B, Goda H, Huang W, Leitzel K, Natori T, Nakano Y, Okada H, Sperinde J, Ali S, Vasekar M, D'Aguiar M, McMahon L, Henry J, Lipton A, Hicks D. ASCO/CAP human epidermal growth factor receptor-2 (HER2) in situ hybridization (ISH) categories evaluated by quantitative HER2 protein diagnostic methodologies: A comparative analysis [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P1-03-02.

A novel detection methodology for HER2 protein quantitation in formalin-fixed, paraffin embedded clinical samples using fluorescent nanoparticles: an analytical and clinical validation study

BackgroundClinical assays for the assessment of the human epidermal growth factor receptor-2 (HER2) status in breast cancer include immunohistochemistry (IHC) and in situ hybridization (ISH), both of which have limitations. Recent studies have suggested that a more quantitative approach to the measurement of HER2 protein expression may improve specificity in selecting patients for HER-2 targeted therapy. In the current study, we have used HER2 expression in breast cancer cell lines and clinical samples as a model to explore the potential utility of a novel immunodetection technique, using streptavidin coated Phosphor Integrated Dot fluorescent nanoparticles (PID), which can be quantitatively measured using computer analysis.MethodsThe expression of HER2 protein in cell lines was evaluated with antibody-binding capacity using fluorescence-activated cell sorting (FACS) for comparison with PID measurements to test for correlations with existing quantitative protein analysis methodologies. Various other analytic validation tests were also performed, including accuracy, precision, sensitivity, robustness and reproducibility. A methods comparison study investigated correlations between PID versus IHC and ISH in clinical samples. Lastly, we measured HER2 protein expression using PID in the pretreatment biopsies from 34 HER2-positive carcinomas that had undergone neoadjuvant trastuzumab-based chemotherapy.ResultsIn the analytic validation, PID HER2 measurements showed a strong linear correlation with FACS analysis in breast cell lines, and demonstrated significant correlations with all aspects of precision, sensitivity, robustness and reproducibility. PID also showed strong correlations with conventional HER2 testing methodologies (IHC and ISH). In the neoadjuvant study, patients with a pathologic complete response (pCR) had a significantly higher PID score compared with patients who did not achieve a pCR (p = 0.011), and was significantly correlated to residual cancer burden (RCB) class (p = 0.026, R2 = 0.9975).ConclusionsAnalytic testing of PID showed that it may be a viable testing methodology that could offer advantages over other experimental or conventional biomarker diagnostic methodologies. Our data also suggests that PID quantitation of HER2 protein may offer an improvement over conventional HER2 testing in the selection of patients who will be the most likely to benefit from HER2-targeted therapy. Further studies with a larger cohort are warranted.

Abstract P2-09-28: New quantitative diagnostic method by fluorescence nanoparticle for HER2 positive breast cancer treated with neoadjuvant lapatinib and trastuzumab: The Neo LaTH study (JBCRG-16TR)

Abstract Background: HER2 (human epidermal growth factor receptor 2) testing performed by IHC (immunohistochemical) methods and FISH (fluorescence in situ hybridization) is semi-quantitative. Exact quantification of HER2 is needed to predict which patients are more or less likely to response to anti HER2 therapy. To improve the method for cancer patients' HER2 status, we developed a novel fluorescence IHC method using new fluorescence nanoparticle. The fluorescent intensity of this new nanoparticles, termed phosphor-integrated dot (PID), was approximately 100-fold brighter than that of Quantum dots. Because of its increased brightness and analyzing technology, this PID-based fluorescent IHC(IHC-PIC) has an ability of quantifying the biomarker protein in the cancer tissue sample at single particle level. In this study, the primary objective was to investigate if pathological complete response (pCR) rate in HER2- positive breast cancer treated by trastuzumab and lapatinib containing neoadjuvant systemic therapy would depend on the level of HER2, EGFR, HER3, Ki67, ER and PgR protein quantified by this new method. Methods: The Neo-LaTH study is a randomized phase II multicenter trial evaluating the efficacy and safety of lapatinib and trastuzumab followed by lapatinib and trastuzumab plus weekly paclitaxel with or without prolongation of anti-HER2 therapy prior to chemotherapy (18 weeks vs. 6 weeks). The primary endpoint was the comprehensive pCR rate. We evaluated the HER2, EGFR, HER3, Ki67, ER and PgR amount by nano-patho method using PID in formalin-fixed paraffin-embedded core biopsy samples taken at diagnosis retrospective analysis. Univariate and multivariate analyses were performed to determine the association between pCR and variables, including HER2, EGFR, HER3, Ki67, ER and PgR nano-patho score and clinicopathological factors including histological grade, tumor status, nodal status and HER2 FISH ratio. Results: A total of 96 tumor samples from patients were used for the present analysis.The pCR rate was 60.4%. We obtained the images of only PID signal by the image analyses, and calculated the number of PID particles in a cell and defined it as IHC-PID score that reflects the level of HER2, EGFR, HER3, Ki67, ER and PgR protein expression in cancer cells. Univariate analysis showed that HER2 IHC-PID score(p&amp;lt;0.0001), ER IHC-PID score(p=0.009) and PgR IHC-PID score(p=0.019) were associated with pCR and multivariate analysis showed that HER2 IHC-PID score was significantly associated with pCR (adjusted odds ratio, 0.990 [95% CI, 0.984–0.996]; P &amp;lt; .0001). Conclusion: We successfully performed the quantitative IHC-PID for HER2, EGFR, HER3, Ki67, ER and PgR. And we propose using HER2 IHC-PID score as a predictive factor for trastuzumab and lapatinib containing neoadjuvant systemic therapy. This quantitative diagnostic method would be expected to contribute to the development of a molecular therapeutic strategy. Citation Format: Tada H, Miyashita M, Gonda K, Watanabe M, Suzuki A, Watanabe G, Harada N, Sato A, Hamanaka Y, Masuda N, Toi M, Ohno S, Bando H, Ishiguro H, Inoue K, Yamamoto N, Kuroi K, Ohuchi N, Ishida T. New quantitative diagnostic method by fluorescence nanoparticle for HER2 positive breast cancer treated with neoadjuvant lapatinib and trastuzumab: The Neo LaTH study (JBCRG-16TR) [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P2-09-28.

The quality of invasive breast cancer care for low reimbursement rate patients: A retrospective study

Though evidence-based treatments have been recommended for breast cancer, underuse of the treatments was still observed. To certain extent, patients’ access to care, which can be enhanced by increasing the coverage of health insurance, could account for the current underuse in recommended care. This study aimed to examine the association between different proportions of reimbursement and quality of recommended breast cancer care, as well as length of hospital stay. In this retrospective study, 3669 patients diagnosed with invasive breast cancer between 1 June, 2011 and 30 June, 2013 were recruited. Seven quality indicators from preoperative diagnosis procedures to adjuvant therapy and one composite indicator were selected as dependent variables. Logistic regression and generalized linear models were used to explore the association between quality of care and length of hospital stay with different reimbursement rates. Compared with UEBMI (urban employment basic medical insurance), which represented high level reimbursement rate, patients with lower rates of reimbursement were less likely to receive core biopsy, HER-2 (human epidermal growth factor receptor-2) testing, BCS (breast conserving surgery), SLNB (sentinel lymph nodes biopsy), adjuvant therapy and hormonal treatment. No significant difference in preoperative length of hospital stay was observed among the three insurance schemes, however URBMI (urban resident basic medical insurance) insured patients stayed longer for total length of hospital stay. Significant disparities in utilization of evidence-based breast cancer care among patients with different proportions of reimbursement were observed. Patients with lower rate of reimbursement were less likely to receive recommended care. Our findings could provide important support for further healthcare reform and quality improvement in breast cancer care.

Human Epidermal Growth Factor Receptor-2 (HER2) expression in Gastric adenocarcinoma and relevant clinicopathological features in a cohort of Sri Lankan patients

Introduction Human epidermal growth factor receptor -2 (HER2) gene over-expression heralds adverse prognosis in gastric adenocarcinoma (GCa). HER2 protein over-expression by immunohistochemistry (IHC) ranges from 10%-22.8% and 11.7%-15.74% in Western and Asian studies respectively. We aimed to evaluate HER2 positivity in GCa by IHC and to study relevant clinicopathological relationships, as information is locally unavailable. Methodology Hundred consecutive GCa (excluding gastrooesophageal junction tumours) were recruited prospectively from the National Hospital of Sri Lanka over two years. The diagnosis was made on endoscopic biopsy/gastrectomy specimens. HER2 was assessed in paraffinized tumour using c-erB-2 oncoprotein (DAKO A0485) and Real-TM Envision system. Scores of 2+ and 3+ were considered positive, using standard HER2 expression criteria. Relationship between HER2 and clinicopathological parameters were analyzed using SPSS-21 and Chi-squared test. Results Study included 44 gastrectomies and 56 biopsies. Mean age was 61.1 years (range 32- 82). HER2 was positive in 9 (9%) GCas: 2+ in 3(5.4%) biopsies and 3(6.8%) resections, 3+ in 1(1.8%) biopsy and 2(4.5%) resections. Eight (88.9%) HER2 positive GCa occurred in =T3) (p=0.31). HER2 positive cases showed, poor differentiation (p=0.047), higher nuclear grade (p=0.029), mitotic counts >5/HPF (p=0.003), extracellular mucin (p=0.05), tumour necrosis (p=0.037), perineural invasion (p=0.002) and nodal metastasis (p=0.04) assessed only in gastric resections. Discussion and conclusion HER2 positivity with immunohistochemistry was 9% in the gastric carcinoma cohort and were associated with higher stage, adverse histological parameters and nodal metastasis in gastric resections. Adverse histological features can be used as screening features for HER2 testing in gastric carcinoma in limited resource settings.

Biomarkers in Pharmaceutical Research.

Biomarkers are important tools in drug development and are used throughout pharmaceutical research. This review focuses on molecular biomarkers in drug development. It contains sections on how biomarkers are used to assess target engagement, pharmacodynamics, safety, and proof-of-concept. It also covers the use of biomarkers as surrogate end points and patient selection/companion diagnostics and provides insights into clinical biomarker discovery and biomarker development/validation with regulatory implications. To survey biomarkers used in drug development--acknowledging that many pharmaceutical development biomarkers are not published--we performed a focused PubMed search employing "biomarker" and the names of the largest pharmaceutical companies as keywords and filtering on clinical trials and publications in the last 10 years. This yielded almost 500 entries, the majority of which included disease-related (approximately 60%) or prognostic/predictive (approximately 20%) biomarkers. A notable portion (approximately 8%) included HER2 (human epidermal growth factor receptor 2) testing, highlighting the utility of biomarkers for patient selection. The remaining publications included target engagement, safety, and drug metabolism biomarkers. Oncology, cardiovascular disease, and osteoporosis were the areas with the most citations, followed by diabetes and Alzheimer disease. Judicious biomarker use can improve pharmaceutical development efficiency by helping to select patients most appropriate for treatment using a given mechanism, optimize dose selection, and provide earlier confidence in accelerating or discontinuing compounds in clinical development. Optimal application of biomarker technology requires understanding of candidate drug pharmacology, detailed modeling of biomarker readouts relative to pharmacokinetics, rigorous validation and qualification of biomarker assays, and creative application of these elements to drug development problems.

HER2 FISH analysis on a skeletal metastasis: a case report and technical review

Eighty per cent of patients who develop metastatic breast cancer will experience a skeletal metastasis. National guidelines recommend biopsy and repeat estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor-2 (HER2) testing for all metastatic breast cancers to guide further treatment planning. Biomarker analysis in bone biopsy samples is challenging due to the necessity of decalcification of the bone sample in order to allow tissue sectioning. The decalcification solutions may result in the degradation of proteins and thus affect the results of immunohistochemical (IHC) testing for ER, PR, and HER2. The decalcification solutions may also degrade nucleic acids, and result in poor or no hybridization signals when attempting in situ hybridization (ISH) testing for HER2. Here, we report a patient with metastatic breast cancer to the bone who underwent image guided core needle biopsy. Her past medical history was significant for ER+, PR+, and HER2+ breast cancer. The biopsy yielded cores of bone, which were decalcified, and a blood clot that was separated from the bone and fixed in formalin without decalcification. Both specimens showed metastatic carcinoma consistent with a breast primary when viewing hematoxylin and eosin (H&E) stained sections. Immunohistochemical analysis, which was performed only on the clot sections, showed ER+, PR − , and HER2 equivocal (2+) tumor cells. Immunohistochemical analysis was not performed on the bony tissue, because our assays are not validated for use on decalcified tissue. Human epidermal growth factor receptor-2 fluorescence in situ hybridization (FISH) analysis was attempted on the bone biopsy and yielded no signals (analytic failure). This is typical of HER2 FISH analysis in decalcified specimens, due to the impaired deoxyribonucleic acid (DNA) quality. However, HER2 FISH analysis on the clot sections showed that the HER2 gene was amplified. Given the clinical importance of repeat HER2 testing in patients with recurrent breast cancer, techniques that avoid decalcification are needed. Separating the blood clot from the bony tissue, and processing it without decalcification may yield viable tumor whose DNA has not been degraded, and it then can be used for the analysis of ER, PR, and HER2.

High HER2 protein levels correlate with increased survival in breast cancer patients treated with anti-HER2 therapy

IntroductionCurrent methods to determine HER2 (human epidermal growth factor receptor 2) status are affected by reproducibility issues and do not reliably predict benefit from anti-HER2 therapy. Quantitative measurement of HER2 may more accurately identify breast cancer (BC) patients who will respond to anti-HER2 treatments. MethodsUsing selected reaction monitoring mass spectrometry (SRM-MS), we quantified HER2 protein levels in formalin-fixed, paraffin-embedded (FFPE) tissue samples that had been classified as HER2 0, 1+, 2+ or 3+ by immunohistochemistry (IHC). Receiver operator curve (ROC) analysis was conducted to obtain optimal HER2 protein expression thresholds predictive of HER2 status (by standard IHC or in situ hybridization [ISH]) and of survival benefit after anti-HER2 therapy. ResultsAbsolute HER2 amol/μg levels were significantly correlated with both HER2 IHC and amplification status by ISH (p < 0.0001). A HER2 threshold of 740 amol/μg showed an agreement rate of 94% with IHC and ISH standard HER2 testing (p < 0.0001). Discordant cases (SRM-MS-negative/ISH-positive) showed a characteristic amplification pattern known as double minutes. HER2 levels >2200 amol/μg were significantly associated with longer disease-free survival (DFS) and overall survival (OS) in an adjuvant setting and with longer OS in a metastatic setting. ConclusionQuantitative HER2 measurement by SRM-MS is superior to IHC and ISH in predicting outcome after treatment with anti-HER2 therapy.

HER2 in Breast Cancer

HER2 (human epidermal growth factor receptor 2) receptor is a membrane tyrosine kinase and when activated affects cell proliferation and survival. The HER2 oncogene is located on chromosome 17q12. HER2 amplification is the primary pathway of HER2 receptor overexpression and is a major driver of tumor development and progression in a subset of breast cancers. HER2 is amplified in about 15% to 20% of breast cancers. The overexpressed HER2 receptor is a valuable therapeutic target. The 2007 ASCO guidelines mandate that HER2 should be evaluated in every invasive breast cancer, either at the time of diagnosis or recurrence to guide therapy. Currently HER2 testing is carried out by several methods. It is crucial to standardize testing techniques to accurately assess HER2 status. The aim of this review on HER2 in breast cancer is to discuss the important aspects of HER2 biology, its significance in breast cancer, and the current standards for its detection.

Central pathology laboratory review of HER2 and ER in early breast cancer: an ALTTO trial [BIG 2-06/NCCTG N063D (Alliance)] ring study

Choice of therapy for breast cancer relies on human epidermal growth factor receptor-2 (HER2) and estrogen receptor α (ER) status. Before randomization in the phase III Adjuvant Lapatinib and/or Trastuzumab Treatment Optimisation (ALTTO) trial for HER2-positive disease, HER2 and ER were centrally reviewed by Mayo Clinic (Rochester, MN, and Scottsdale, AZ) for North America and by the European Institute of Oncology (IEO; Milan, Italy) for the rest of world (except China). Discordance rates (local vs. central review) differed between Mayo and IEO. Among locally HER2-positive cases, 5.8 % (Mayo) and 14.5 % (IEO) were centrally HER2 negative. Among locally ER-positive cases, 16.2 % (Mayo) and 4.2 % (IEO) were centrally ER-negative. Among locally ER-negative cases, 3.4 % (Mayo) and 21.4 % (IEO) were centrally ER-positive. We, therefore, performed a ring study to identify features contributing to these differing discordance rates. Mayo and IEO exchanged slides for 25 HER2 and 35 ER locally/centrally discordant cases. Both laboratories performed IHC and FISH for HER2 using the HercepTest(®) and PathVysion HER2 DNA probe kit/HER2/centromere 17 probe mixture. IHC for ER was tested centrally using the monoclonal ER 1D5 antibody (Mayo) or the DAKO cocktail of ER 1D5 and 2.123 antibodies (IEO). Mayo and IEO confirmed the central HER2-negative result in 100 % of 25 cases. Mayo and IEO confirmed the central ER result in 29 (85 %) of 34 evaluable cases. The five Mayo-negative/IEO-positive cases were ER-positive when retested at Mayo using the DAKO ER cocktail. In this ring study, ALTTO ineligibility did not change when HER2 testing was performed by either IEO or Mayo central laboratories. However, a dual antibody ER assay had fewer false-negative test results than an assay with a single antibody, and there was more discordance between the two ER reagents than has been previously reported. Using even slightly different assay methods yielded different results, even between experienced central laboratories.

The evolving role of HER2 evaluation for diagnosis and clinical decision making for breast and gastric adenocarcinoma

Clinical laboratory testing for human epidermal growth factor receptor-2 (HER2) in patients with newly diagnosed breast and gastric cancer is critically important for therapeutic decisions about adjuvant treatment. The HER2 pathway is a major molecular driver of disease progression in a subset of these solid tumors and the results of HER2 testing determine which patients are likely to respond to an expanding variety of therapies that target this important biologic pathway. Given the significant clinical impact of these test results on patient management and adjuvant treatment decisions, standardization of the assay for HER2 and assured reliability of these results are critical. We review highlights of the clinical rationale that underlies HER2 testing for both breast and gastric carcinomas, and describe some of the challenges associated with ensuring accurate test results.