Human Epidermal Growth Factor Receptor-2 Gene Research Articles

Her two Gene Amplification by Fish Technique in Ovarian Cancer Patients

Human Epidermal Growth Factor Receptor-2 (HER2) gene triggers a cascade of cellular responses leading to cell proliferation, survival, and metastasis in malignant cells. Therapies targeting HER2 gene are emerging as a promising treatment option for patients with HER2 gene amplification. Hence, the present study aims to investigate the HER2 gene amplification and relationship with clinicopathological features in ovarian cancer patients.

PET/CT and SPECT/CT Imaging of HER2-Positive Breast Cancer.

HER2 (Human Epidermal Growth Factor Receptor 2)-positive breast cancer is characterized by amplification of the HER2 gene and is associated with more aggressive tumor growth, increased risk of metastasis, and poorer prognosis when compared to other subtypes of breast cancer. HER2 expression is therefore a critical tumor feature that can be used to diagnose and treat breast cancer. Moving forward, advances in HER2 in vivo imaging, involving the use of techniques such as positron emission tomography (PET) and single-photon emission computed tomography (SPECT), may allow for a greater role for HER2 status in guiding the management of breast cancer patients. This will apply both to patients who are HER2-positive and those who have limited-to-minimal immunohistochemical HER2 expression (HER2-low), with imaging ultimately helping clinicians determine the size and location of tumors. Additionally, PET and SPECT could help evaluate effectiveness of HER2-targeted therapies, such as trastuzumab or pertuzumab for HER2-positive cancers, and specially modified antibody drug conjugates (ADC), such as trastuzumab-deruxtecan, for HER2-low variants. This review will explore the current and future role of HER2 imaging in personalizing the care of patients diagnosed with breast cancer.

A Transcriptional Link between HER2, JAM-A and FOXA1 in Breast Cancer.

Overexpression of the human epidermal growth factor receptor-2 (HER2) is associated with aggressive disease in breast and certain other cancers. At a cellular level, the adhesion protein Junctional Adhesion Molecule-A (JAM-A) has been reported to regulate the expression of HER3 via a transcriptional pathway involving FOXA1. Since FOXA1 is also a suggested transcription factor for HER2, this study set out to determine if JAM-A regulates HER2 expression via a similar mechanism. An integrated tripartite approach was taken, involving cellular expression studies after targeted disruption of individual players in the putative pathway, in silico identification of relevant HER2 promoter regions and, finally, interrogation of cancer patient survival databases to deconstruct functionally important links between HER2, JAM-A and FOXA1 gene expression. The outcome of these investigations revealed a unidirectional pathway in which JAM-A expression transcriptionally regulates that of HER2 by influencing the binding of FOXA1 to a specific site in the HER2 gene promoter. Moreover, a correlation between JAM-A and HER2 gene expression was identified in 75% of a sample of 40 cancer types from The Cancer Genome Atlas, and coincident high mean mRNA expression of JAM-A, HER2 and FOXA1 was associated with poorer survival outcomes in HER2-positive (but not HER2-negative) patients with either breast or gastric tumors. These investigations provide the first evidence of a transcriptional pathway linking JAM-A, HER2 and FOXA1 in cancer settings, and support potential future pharmacological targeting of JAM-A as an upstream regulator of HER2.

Blood digital polymerase chain reaction as a potential method to detect human epidermal growth factor receptor 2 amplification in non-small cell lung cancer

BackgroundThis study aimed to verify the feasibility of human epidermal growth factor receptor-2 (HER2) amplification detection by digital polymerase chain reaction (dPCR) in non-small cell lung cancer (NSCLC) patients and explore whether HER2 amplification could be detected in circulating tumor DNA (ctDNA) by dPCR.MethodsA total of 112 fresh biopsy tissues and 88 blood samples from NSCLC patients were collected. The serum ctDNA was obtained from blood samples. The copy number of the HER2 gene was evaluated by dPCR and next-generation sequencing (NGS). The sensitivity/specificity and survival analysis were performed by the receiver operating characteristic (ROC) curve. The survival analysis was performed by Kaplan-Meier (KM) curve and univariate Cox regression analysis was also conducted.ResultsROC analysis showed a good prediction result for HER2 amplification in blood samples by dPCR. The survival analysis showed that the median overall survival (OS) in the HER2 negative group detected by blood dPCR was significantly different from the positive group. The results of multivariate Cox regression were the same as those of survival analysis.ConclusionsBlood dPCR might be a potential method to detect HER2 amplification in NSCLC. Amplification of the HER2 gene detected by dPCR was correlated with OS in NSCLC.

Detection of Genetic Polymorphism of HER2 Gene in HER2 Positive Breast Cancer Women in Iraq

The human epidermal growth factor receptor-2 (HER2) gene plays a critical role in breast cancer development and progression. HER2 overexpression characterizes a biologically and clinically aggressive breast cancer subtype. In this study, 60 samples from Iraqi women with breast cancer were collected and investigated for HER2 protein in the tissue by immunohistochemistry. Also, 20 samples from healthy Iraqi women were used as a control. The results showed that 18 (30 %) patients expressed the HER2 protein. A molecular study for single nucleotide polymorphism (SNP) was conducted on samples metastasizing to lymph nodes. DNA was extracted and polymerase chain reaction (PCR) was performed to amplify exon 17 and intron 17 of HER2 gene. Sequencing of PCR product was achieved and two SNPs of HER2 gene, one in exon 17 (Ile655Val) and another close to it in intron 17 (rs903506) were studied. In exon 17, SNP Ile655Val was found in 41% of patients, while in intron 17, the non-coding SNP rs903506 was found in 27% of patients. However, no polymorphism was found in the control group. The results may suggest that HER2 gene can be used as a molecular marker for breast cancer.

The Exciting New Field of HER2-Low Breast Cancer Treatment.

Simple SummaryBreast cancer can express, at varied levels, a protein named HER2, commonly responsible for making it grow and send distant metastases. In the past, patients affected by the so called HER2-positive breast cancer had lower probabilities of cure and survival, though with the advent of drugs that target HER2, three decades ago, their prognosis has greatly improved. So far, only patients with strong HER2 expression on their tumour can be treated with these benefitial drugs, like trastuzumab, though recently stronger drugs have also been shown capable of eliminating breast cancer cells with lower levels of HER2 expression (HER2-low). Sooner or later, these new drugs, like trastuzumab-deruxtecan, may be available for treating such patients. Therefore, the aim of this narrative review of the literature is to provide an outline of what is going on on this specific field of research, and what could be expected in the future in the clinic. Since human epidermal growth factor receptor-2 (HER2) characterization, going through clinical research and regulatory approval of HER2-targeted therapies, much has elapsed and is still unfolding. Hitherto, only breast cancer (BC) patients with HER2 immunohistochemistry 3+ or with HER2 gene fluorescence in-situ hybridization (FISH) amplification (a.k.a., HER2-positive BC) have benefited from anti-HER2 agents. In recent years, however, much of the research effort has been expanded, with positive outcomes being reached for formerly known HER2-negative BC that yet express HER2 to some degree (HER2 immunohistochemistry 1+ or 2+, but FISH negative) and are currently being classified as HER2-low BC for the purpose of trial enrollment. In this sense, our aim is to review the body of evidence of HER2-low BC that led to the study of first-generation anti-HER2 agents, like trastuzumab, and how they have failed to achieve any clinical applicability in this setting. In addition, we review new data that is leading to the growing success of the new generation of drugs, especially the promising HER2-directed antibody–drug conjugates. A narrative review is also performed regarding the rationale behind the consolidated and ongoing clinical trials studying anti-HER2 agents in combination with unrelated agents, such as immunotherapy, endocrine therapy, and CDK4/6 inhibitors. Hopefully, all this ongoing research effort will be able to extend the survival benefits seen with anti-HER2 agents in HER2-positive disease, at least to some degree, to the greater proportion of patients with HER2-low BC.

Research Progress of Targeted Therapy for HER2 Gene in Advanced Non-small Cell Lung Cancer

肺癌是最常见的恶性肿瘤，非小细胞肺癌（non-small cell lung cancer, NSCLC）发生率约占所有肺癌的85%。人表皮生长因子受体2（human epidermal growth factor receptor-2, HER2）是ERBB/HER家族中的一种酪氨酸激酶受体，与表皮生长因子受体（epidermal growth factor receptor, EGFR）等其他家族成员一起激活下游信号传导。HER2基因突变与许多上皮细胞癌症的恶化程度密切相关，HER2基因突变的肿瘤表现出较强的转移能力和侵润能力，对化疗的敏感性也较差，且易复发。目前肺癌驱动基因靶向治疗进展迅速。作为驱动基因，NSCLC中HER2基因突变频率相对于EGFR较低，但是其肺癌驱动机制明确且部分靶向治疗有效，将来可能成为新的标准治疗手段。本文重点讨论HER2基因突变在NSCLC治疗中的研究进展。

Amplification and clinicopathological significance of HER-2 in Kazakh esophageal squamous cell carcinoma.

Amplification and overexpression of the human epidermal growth factor receptor-2 (HER-2) gene accelerates cell division and proliferation, and promotes tumor growth and metastasis in various malignant tumors. However, there are few reports on its influence and mechanism in esophageal cancer. The aim of the present study was to investigate the gene amplification and clinicopathological significance of HER-2 in Kazakh esophageal squamous cell carcinoma (ESCC). HER-2 gene amplification was detected in 70 esophageal cancer tissues using fluorescence in situ hybridization. The association between the HER-2 gene amplification and the clinicopathological characteristics of patients with esophageal cancer was also analyzed. The amplification rate of the HER-2 gene in patients with esophageal cancer was 54.2% (38/70). The results also revealed a positive association between the amplification rate of the HER-2 gene in esophageal squamous cell carcinomas and the level of tissue differentiation, increasing gradually and significantly among the highly, moderately and poorly differentiated tissues (P<0.05). The amplification rate of the HER-2 gene in patients with lymph node metastasis was higher than those without (P<0.05). There was no significant association between the amplification rate of the HER-2 gene and any of the clinic pathological parameters, such as sex, age, depth of invasion and 3-year survival, among patients (P>0.05). In conclusion, the amplification rate of the HER-2 gene in patients with Kazakh ESCC was high. There was an association with various prognostic factors, including cancer differentiation and lymph node metastasis. HER-2 gene expression levels may be considered as an indicator of poor prognosis in patients with ESCC in the clinical setting, and this may provide a basis of treatment for individualized targeted therapies.

Somatic Mutations in HER2 and Implications for Current Treatment Paradigms in HER2-Positive Breast Cancer.

In one of every four or five cases of breast cancer, the human epidermal growth factor receptor-2 (HER2) gene is overexpressed. These carcinomas are known as HER2-positive. HER2 overexpression is linked to an aggressive phenotype and a lower rate of disease-free and overall survival. Drugs such as trastuzumab, pertuzumab, lapatinib, neratinib, and the more recent afatinib target the deregulation of HER2 expression. Some authors have attributed somatic mutations in HER2, a role in resistance to anti-HER2 therapy as differential regulation of HER2 has been observed among patients. Recently, studies in metastatic ER + tumors suggest that some HER2 mutations emerge as a mechanism of acquired resistance to endocrine therapy. In an effort to identify possible biomarkers of the efficacy of anti-HER2 therapy, we here review the known single-nucleotide polymorphisms (SNPs) of the HER2 gene found in HER2-positive breast cancer patients and their relationship with clinical outcomes. Information was recompiled on 11 somatic HER2 SNPs. Seven polymorphisms are located in the tyrosine kinase domain region of the gene contrasting with the low number of mutations found in extracellular and transmembrane areas. HER2-positive patients carrying S310F, S310Y, R678Q, D769H, or I767M mutations seem good candidates for anti-HER2 therapy as they show favorable outcomes and a good response to current pharmacological treatments. Carrying the L755S or D769Y mutation could also confer benefits when receiving neratinib or afatinib. By contrast, patients with mutations L755S, V842I, K753I, or D769Y do not seem to benefit from trastuzumab. Resistance to lapatinib has been reported in patients with L755S, V842I, and K753I. These data suggest that exploring HER2 SNPs in each patient could help individualize anti-HER2 therapies. Advances in our understanding of the genetics of the HER2 gene and its relations with the efficacy of anti-HER2 treatments are needed to improve the outcomes of patients with this aggressive breast cancer.

HER2 Heterogeneity in Personalized Therapy of Gastro-Oesophageal Malignancies: An Overview by Different Methodologies.

Human epidermal growth factor receptor-2 (HER2)-expression gastro-oesophageal adenocarcinomas (GEA) gained interest as an important target for therapy with trastuzumab. In the current review, we focused the current knowledge on HER2 status in dysplastic and neoplastic gastric conditions, analyzing the methodological procedures to identify HER2 expression/amplification, as well as the proposed scoring recommendations. One of the most relevant questions to evaluate the useful impact of HER2 status on therapeutic choice in GEAs is represented by the significant heterogeneity of HER2 protein and gene expression that may affect the targeted treatment selection. Future development of biotechnology will continue to evolve in order to offer more powerful detection systems for the assessment of HER2 status. Finally, liquid biopsy as well as mutation/amplification of several additional genes may furnish an early detection of secondary HER2 resistance mechanisms in GEAs with a better monitoring of the treatment response.

Human Epidermal Growth Factor Receptor-2 gene polymorphism and breast cancer risk in women from the Northeastern region of Brazil

OBJECTIVES:In the Human Epidermal Growth Factor Receptor-2 (HER2) rs1136201 variant, the presence of the G allele may promote cellular alterations and increase breast cancer risk, in addition to enhanced cellular proliferation, tumor aggressiveness, and metastases. The aim of this study was to investigate the presence of the single-nucleotide polymorphism (SNP) variant, rs1136201, within the HER2 gene in women from the Northeastern region of Brazil and breast cancer risk.METHODS:The study included 140 women who were divided into two groups, case (breast cancer) and control (without breast cancer), with 70 women in each group. Peripheral blood of each woman was drawn for the study of genomic Deoxyribonucleic acid (DNA) extracted from leukocytes using the genotyping technique by real-time polymerase chain reaction.RESULTS:The GG genotype occurred in 1 woman in both groups (1.4%) (p=0.32), while the AG genotype occurred in 19 (27.2%) and 13 (18.6%) women in the case and control (p=1.00) groups, respectively. No statistically significant difference in GG and AG genotypes was observed between the case and control groups in premenopausal women (p=1.00). Furthermore, no significant difference in genotypes was observed between the groups, among postmenopausal women (p=0.14).CONCLUSION:In this study, the HER2 rs1136201 polymorphism did not show any statistically significant association with breast cancer, both in premenopausal and postmenopausal women. Nevertheless, further studies with a larger sample size should be performed to assess the association of HER2 polymorphism with breast cancer risk in women from the Northeastern region of Brazil.

276P - Overexpression of HER3 and the survival of ovarian cancer

Background: Human epidermal growth factor receptor-2 (HER2) and 3 (HER3) belongs to epidermal growth factor receptor (EGFR) family of transmembrane receptor tyrosine kinases which is responsible for cell survival and proliferation, but their role in ovarian cancer is not yet established. In this study, their status in specimens of epithelial ovarian cancer was determined with immunohistochemistry (IHC) and in situ hybridization (ISH) and was analyzed to find out the correlation with clinical features of ovarian cancer. Methods: Tissue microarrays (TMAs) were prepared from paraffin blocks of 105 ovarian tumor samples. The immunohistochemical expression of HER2, HER3, PI3K, Akt, p-Akt, mTOR, p-mTOR, S6 and p-S6 were investigated. HER2 and HER3 gene amplifications were evaluated by ISH. The correlation between status of materials and disease outcome of the patients were analyzed. Results: HER2 positivity was 3.8% on IHC and 5.7% on ISH, otherwise HER3 showed 12.4% and 8.6% expression respectively. The concordances of IHC and ISH were 98.1% (103/105) in HER2, and 82.9% (87/105) in HER3. HER2 status was not related with disease outcome of ovarian cancer. However, the HER3 status on FISH was associated with rapid disease progression, and this was a significant risk factor even after adjusting other possible risk factors in multivariate analysis. Conclusions: HER3 could be an independent marker for poor prognosis in ovarian cancer, because HER3 signaling pathway might be distinguished from that of HER2. The possibility of target therapy in patients with HER3 gene alteration should be evaluated in the field of ovarian cancer. Legal entity responsible for the study: The authors. Funding: Has not received any funding. Disclosure: All authors have declared no conflicts of interest.

HER2 status in patients with residual rectal cancer after preoperative chemoradiotherapy: the relationship with molecular results and clinicopathologic features.

The specific role of human epidermal growth factor receptor-2 (HER2) status in rectal cancers remains unclear. This study therefore aimed to explore clinicopathologic and molecular characteristics, and prognostic value of HER2-positivity in residual mid- and/or low-rectal cancers after preoperative chemoradiotherapy (CRT). Surgical specimens from 145 patients with residual rectal cancer after preoperative CRT between January 2006 and January 2011 were used to evaluate HER2 status. HER2 protein expression and gene amplification were determined using immunohistochemistry (IHC) and silver in situ hybridization (SISH) on whole tissue sections, respectively. Polymerase chain reaction was used to analyze molecular characteristics, including microsatellite instability (MSI) and mutations in KRAS exon 2 (codon 12 and 13) and BRAF V600E mutation. Of 139 eligible patients, 8 (5.8%) had HER2 overexpression (IHC 2+ and 3+) that was not associated with clinicopathologic characteristics and patient survival, except positive circumferential resection margin (CRM) (P = 0.012). SISH was performed on 24 patient samples with IHC 1+ (n = 16), 2+ (n = 6), and 3+ (n = 2). HER2 amplification was identified in 3 patients (2.2%); however, this was also associated with positive CRM (P = 0.009) but not survival (all P > 0.05). Moreover, HER2 overexpression and amplification had no relationship with KRAS or BRAF mutations, and MSI status (all P > 0.05). HER2 positivity was found in a minority of rectal cancer patients and was not significantly associated with clinicopathologic and molecular characteristics. Our findings can be helpful in understanding the clinicopathologic bases of HER2 status in rectal cancers.

Abstract P3-08-16: ER, PR and HER2 biomarkers in UK and Irish clinical breast cancer testing: analysis of results from &amp;gt;168,000 patients

Abstract AIMS To describe and analyse &amp;gt;168,000 sets of results from clinical breast biomarker testing carried-out between 2009 and 2016 in the UK and Republic of Ireland, focussing on biological relationships. To present robust confirmatory evidence on known associations and provide new data on previously undescribed or unconfirmed ones. BACKGROUND Between Jan-2009 and Jan-2016 the UK National External Quality Assessment Scheme for Immunocytochemistry and In-situ Hybridisation (UK NEQAS ICC&amp;ISH) collected results on clinical breast cancer testing, comprehensively for human epidermal growth factor receptor-2 (HER2) and optionally for estrogen receptor (ER) and progesterone receptor (PR), from most UK and Irish testing centres. Primary objectives were to assess and, where indicated achieve improvements in testing quality. The size and scope of the data-set is unparalleled in the literature and represent a significant reference resource for the breast cancer community. METHODS UK NEQAS ICC&amp;ISH created and curated an on-line data-entry system allowing centres to systematically collect their own HER2, ER and PR results from clinical testing; for HER2, immunohistochemical (IHC) and in-situ hybridisation (ISH) data were collected; for ER and PR, IHC results could be entered according to local practise format. Tools and guidance were provided enabling centres to analyse their data for local audit and quality assurance. Clinicopathologic data was also collected, including: patient age at diagnosis, histological tumor type, tumor grade, site/stage (primary, recurrence, metastasis) and sample type (core, excision). RESULTS Data are present on 168,793 patients. 173 centres contributed ≥100 entries (96% of total). Median age was 62 years (IQR: 51-72). Tumor type was stated in 42%: 76% were invasive ductal, and 13% invasive lobular carcinoma. Grade was stated in 56%: 15% were Grade 1, 55% Grade 2 and 30% Grade 3. Site/stage was stated in 56%: 92% were primary, 2% recurrent and 6% metastatic. Sample type was stated in 70%: 75% were cores and 25%, excision. Receptor statuses were available as follows; HER2 (100%): 87% negative, 13% positive; ER (45%): 15% negative, 85% positive; PR (31%): 29% negative, 71% positive. HER2 data were available as follows; category by IHC: 91%; amplification status by ISH: 15%; HER2 gene/chromosome 17 (CEP17) ratio: 15% (86% of which were IHC 2+); HER2 gene copy number: 7%; CEP17 copy number: 7%. HER2-positive rate was 24% in ER-negative and 11% in ER-positive cases. HER2-positive status was negatively associated with increasing ER positivity (rho -0.22, P&amp;lt;0.001). 71% of HER2-positive cases were ER-positive and 29% were ER-negative. The HER2 2+ rate was 14% in ER-negative and 21% in ER-positive cases. Considering only IHC 2+ cases, median HER2 copy number was 5.4 in ER-negative and 4.4 in ER-positive disease. Comprehensive description of significant associations for all parameters will be presented. CONCLUSION The unique size and scope of this data-set has allowed confirmation of known associations for HER2, ER and PR with clinicopathologic and biological characteristics in breast cancer to very high confidence levels, and has uncovered previously undescribed relationships in both ER-positive and ER-negative disease. Citation Format: Dodson A, Parry S, Ibrahim M, Bartlett J, Dowsett M, Miller K. ER, PR and HER2 biomarkers in UK and Irish clinical breast cancer testing: analysis of results from &amp;gt;168,000 patients [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 P3-08-16.

A novel HER2 gene body enhancer contributes to HER2 expression

The transcriptional regulation of the human epidermal growth factor receptor-2 (HER2) contributes to an enhanced HER2 expression in HER2-positive breast cancers with HER2 gene amplification and HER2-low or HER2-negative breast cancers following radiotherapy or endocrine therapy, and this drives tumorigenesis and the resistance to therapy. Epigenetic mechanisms are critical for transcription regulation, however, such mechanisms in the transcription regulation of HER2 are limited to the involvement of tri-methylated histone 3 lysine 4 (H3K4me3) and acetylated histone 3 lysine 9 (H3K9ac) at the HER2 promoter region. Here, we report the identification of a novel enhancer in the HER2 3’ gene body, which we have termed HER2 gene body enhancer (HGE). The HGE starts from the 3’ end of intron 19 and extends into intron 22, possesses enhancer histone modification marks in specific cells and enhances the transcriptional activity of the HER2 promoters. We also found that TFAP2C, a known regulator of HER2, binds to HGE and is required for its enhancer function and that DNA methylation in the HGE region inhibits the histone modifications characterizing enhancer and is inversely correlated with HER2 expression in breast cancer samples. The identification of this novel enhancer sheds a light on the roles of epigenetic mechanisms in HER2 transcription, in both HER2-positive breast cancer samples and individuals with HER2-low or HER2-negative breast cancers undergoing radiotherapy or endocrine therapy.

Abstract P1-03-10: HER2 expression in clinical breast cancer samples: A novel detection methodology for HER2 protein quantitation using fluorescent nanoparticles

Abstract Background: The human epidermal growth factor receptor-2 (HER2) is a member of a family of transmembrane tyrosine kinase receptors that play an important role in regulated normal cell growth and differentiation. The over-expression of HER2 in a subset of 15-20% of invasive breast cancers has an important bearing on prognosis, as HER2-positive tumors are associated with an aggressive clinical course and poor outcome. Targeting HER2-overexpression has been shown to be a remarkably effective therapeutic modality; however testing of tumor samples to assess the HER2 status of the patient's breast cancer is required. Clinical assays to assess the HER2 status in patients being considered for targeted therapy include immunohistochemistry (IHC), which detects protein over-expression, or fluorescence in situ hybridization (FISH), which detects gene amplification. Both the IHC and FISH methodologies have limitations. Given that the target of the currently approved drugs is the receptor protein, novel detections systems that could more accurately and quantitatively detect HER2 protein in clinical samples over a broad dynamic range would be advantageous and may be clinically helpful. Material and Methods: A novel detection technology using streptavidin-coated Phosphor Integrated Dot fluorescent nanoparticles (PID) has been developed that can be visualized by fluorescence microscopy and used for quantitative immunofluorescence detection of protein in clinical samples using computer assisted image analysis. In the current study, PID- nanoparticles were used to analyze HER2 protein expression in breast cancer cell lines and 120 well characterized breast cancer samples. These results have been compared with HER2 IHC and HER2 FISH analysis. Results: The expression levels of HER2 protein from 8 breast cancer cell lines was evaluated by antibody-binding capacity with FACS analysis. Formalin fixed paraffin embedded cell pellets for these cell lines were prepared and used for quantitative HER2 analysis by PID. The PID score/cell for each of these cell lines showed a strong linear correlation with antibody-binding capacity sites/cell by FACS analysis (R2 = 0.94). For the 120 breast cancer samples, PID score/cell was measured and compared against HER2 IHC membrane intensity measure by image analysis (Aperio) and HER2 FISH results. The HER2 PID score/cell showed a correlation coefficient of R2=0.72 versus the average HER2 copy number per cell by FISH, compared with a correlation coefficient of R2=0.41 for HER2 IHC membrane intensity measured by Aperio. For the HER2/CEP17 ratio, the correlation coefficient for the PID score/cell was R2=0.79 compared with a correlation coefficient of R2=0.32 for the HER2 IHC membrane intensity. Conclusions: PID-nanoparticles demonstrate great potential for the quantitative measurement of protein of clinical interest in routine clinical samples with morphologic confirmation of the tissue being studied. Further studies looking for PID-score thresholds for HER2 gene amplification and correlations with clinical outcome data are warranted and ongoing. Citation Format: Hicks DG, Goda H, Zhai H, Okada H, McMahon L, Sullivan N, Tang P, Nakano Y. HER2 expression in clinical breast cancer samples: A novel detection methodology for HER2 protein quantitation using fluorescent nanoparticles [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P1-03-10.

Development of a quantitative PCR assay to evaluate HER2 status of Gastric carcinoma in a cohort of Sri Lankan patients

Introduction Human epidermal growth factor receptor2 (HER2) protein over expression and/or HER2gene amplification are/is linked to dismal outcome of Gastric carcinoma (GCa). Immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) are key-methods to identify patients for HER2 targeted therapy. Drawbacks of both methods warrant novel-tests. Objective To determine whether quantitative Polymerase Chain Reaction (qPCR) could serve as a supplementary-method to evaluate HER2 status of GCa in a cohort of Sri Lankan patients and investigate correlation between HER2 assessed by different methods and clinicopathological features. Methods Twenty GCa-patients with known IHC-HER2 scores were evaluated. qPCR was performed for HER2 gene and Ameloid precursor protein (reference gene) in Formalin fixed paraffin embedded GCa tissue. Threshold values (Ct) were analyzed using Pfaffl-method to detect HER2 gene amplification. Results HER2positivity by IHC (protein) and qPCR (gene) were 20% and 35% respectively. Sensitivity and specificity of qPCR was 67% and 76% respectively and results were reproducible. HER2 protein positivity was correlated with Tumour TNM-stage and Lauren-histological types (P Discussion & Conclusion Discrepancies between HER2 positivity by IHC and qPCR were possibly due to transcription activation by other genes in the absence of HER2 gene amplification and the aberrant form of HER2 protein not detected by IHC. Studies also indicate a higher-proportion of IHC negative, but HER2gene amplified GCa by FISH. qPCR may be used as a supplementary-method for detection of HER2 status of GCa in local setting.

Application of a quantitative PCR assay to evaluate Human Epidermal Growth Factor Receptor - 2 status of Gastric carcinoma in a cohort of Sri Lankan patients

Introduction Human epidermal growth factor receptor2 (HER2) protein over expression and/or HER2 gene amplification is linked to dismal outcome of gastric carcinoma (GCa). Immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) are key methods to identify patients for HER2 targeted therapy. Draw backs of both methods warrant novel tests. The objectives were to determine whether quantitative Polymerase Chain Reaction (qPCR) could serve as a supplementary method to evaluate HER2 status of GCa and investigate the correlation between HER2 assessed by different methods and clinicopathological features. Methodology Twenty GCa patients with known IHC-HER2 scores were evaluated. qPCR was performed for HER2 gene and Amyloid precursor protein (reference gene) in formalin fixed paraffin embedded GCa tissue. Threshold values (Ct) were analyzed using Pfaffl-method to detect HER2 gene amplification. Results HER2 positivity by IHC (protein) and qPCR (gene) were 20% and 35% respectively. Sensitivity and specificity of qPCR was 67% and 76% respectively and results were reproducible. HER2 protein positivity was correlated with TNM-stage and Lauren histological types (P Discussion Discrepancies between HER2 positivity by IHC and qPCR were possibly due to transcription activation by other genes in the absence of HER2 gene amplification and the aberrant form of HER2 protein not detected by IHC. Studies also indicate a higher proportion of IHC negative, but HER2 gene amplified GCa by FISH. Conclusion qPCR may be used as a supplementary method for evaluation of HER2 status in gastric carcinoma proven to be HER2 negative by immunohistochemistry.

Genetic Heterogeneity of HER2 Amplification and Telomere Shortening in Papillary Thyroid Carcinoma.

Extensive research is dedicated to understanding if sporadic and familial papillary thyroid carcinoma are distinct biological entities. We have previously demonstrated that familial papillary thyroid cancer (fPTC) cells exhibit short relative telomere length (RTL) in both blood and tissues and that these features may be associated with chromosome instability. Here, we investigated the frequency of HER2 (Human Epidermal Growth Factor Receptor 2) amplification, and other recently reported genetic alterations in sporadic PTC (sPTC) and fPTC, and assessed correlations with RTL and BRAF mutational status. We analyzed HER2 gene amplification and the integrity of ALK, ETV6, RET, and BRAF genes by fluorescence in situ hybridization in isolated nuclei and paraffin-embedded formalin-fixed sections of 13 fPTC and 18 sPTC patients. We analyzed BRAFV600E mutation and RTL by qRT-PCR. Significant HER2 amplification (p = 0.0076), which was restricted to scattered groups of cells, was found in fPTC samples. HER2 amplification in fPTCs was invariably associated with BRAFV600E mutation. RTL was shorter in fPTCs than sPTCs (p < 0.001). No rearrangements of other tested genes were observed. These findings suggest that the association of HER2 amplification with BRAFV600E mutation and telomere shortening may represent a marker of tumor aggressiveness, and, in refractory thyroid cancer, may warrant exploration as a site for targeted therapy.

HER-2 expression and clinical significance in esophagogastric junction adenocarcinoma (Siewert type Ⅱ)

To investigate the human epidermal growth factor receptor-2 (HER-2) expression in esophagogastirc junction adenocarcinoma(Siewert type Ⅱ)and its clinical significance. A total of 180 patients with esophagogastric junction adenocarcinoma (Siewert type Ⅱ) were included in this study. The HER-2 expression was detected by immunohistochemistry (IHC), and fluorescence in situ hybridization (FISH) analysis was performed to assess the HER-2 gene amplification in the IHC-positive and IHC-weak positive cases. HER-2 overexpression (3+ ), weak positive (2+ ) and negative (1+ /0) was 11.7%(21/180), 8.9%(16/180), and 79.4%(143/180), respectively. The FISH analysis showed HER-2 gene amplification in 95.2%(20/21) of HER-2(3+ ) cases and 18.8% (3/16) of HER-2(2+ ) cases. The concordance rate between IHC and FISH was 95.2%. Overexpression of HER-2(3+ ) was associated with the tumor differentiation (P<0.05), and irrelevant to age, sex, pT stage, pN stage, pM stage and pTNM stage (P>0.05). The median overall survival time (OS) was 13 months in HER-2(3+ ) patients, significantly shorter than the 21 months in HER-2(2+ ) and HER-2(+ /-) cases (P<0.01). Approximately 11.7% of the patients with esophagogastric junction adenocarcinoma (Siewert type Ⅱ) are HER-2-positive on IHC. HER-2 overexpression is associated with the tumor differentiation. IHC can be used as a screening test for the positive expression of HER-2 in the esophagogastirc junction adenocarcinoma (Siewert type Ⅱ). However, FISH detection can be used as a more reliable detection method.