Overexpression Of Human Epidermal Growth Factor Receptor-2 Research Articles

Targeting HER2 in colorectal cancer

Among the molecular subgroups of interest in metastatic colorectal cancer (mCRC), innovations are underway for tumors with overexpression of HER2 (Human Epidermal Growth Factor Receptor 2). Overexpression of the HER2 protein concerns 2 to 5% of CRC at any stage mainly located in the distal colon and rectum. Diagnosis is based on immunohistochemistry, in situ hybridization with appropriate criteria for colorectal localization, and molecular biology (NGS: next-generation sequencing). Overexpression of HER2 is a predictive factor for resistance to treatments targeting EGFR which are indicated in the case where the tumor is wild-type RAS. It seems to be associated with a poor prognosis of mCRC with a higher risk of brain metastasis. Regarding treatments targeting HER2, no randomized controlled phase III has been published to date. However, several combinations have been evaluated in phase II with clinically meaningful objective response rates: trastuzumab-deruxtecan (45%), trastuzumab-tucatinib (46%), trastuzumab-pyrotinib (45%), trastuzumab-pertuzumab (30%) ou trastuzumab-lapatinib (30%). In this literature review, we present here the current state of knowledge on the diagnostic methods of HER2 overexpression in CRC, the main clinical, molecular and prognostic characteristics, and the efficacy results of the different therapeutic combinations for the patients with HER2 overexpressed mCRC. This justifies, despite the lack of marketing authorization in France and in Europe for agents targeting HER2 in CRC, the systematic evaluation of the HER2 status, as recommended in particular by the NCCN (National Comprehensive Cancer Network).

The production of the first functional antibody mimetic in higher plants: the chloroplast makes the DARPin G3 for HER2 imaging in oncology

BackgroundDesigned mimetic molecules are attractive tools in biopharmaceuticals and synthetic biology. They require mass and functional production for the assessment of upcoming challenges in the near future. The DARPin family is considered a mimetic pharmaceutical peptide group with high affinity binding to specific targets. DARPin G3 is designed to bind to the HER2 (human epidermal growth factor receptor 2) tyrosine kinase receptor. Overexpression of HER2 is common in some cancers, including breast cancer, and can be used as a prognostic and predictive tool for cancer. The chloroplasts are cost-effective alternatives, equal to, and sometimes better than, bacterial, yeast, or mammalian expression systems. This research examined the possibility of the production of the first antibody mimetic, DARPin G3, in tobacco chloroplasts for HER2 imaging in oncology.ResultsThe chloroplast specific DARPin G3 expression cassette was constructed and transformed into N. tabacum chloroplasts. PCR and Southern blot analysis confirmed integration of transgenes as well as chloroplastic and cellular homoplasmy. The Western blot analysis and ELISA confirmed the production of DARPin G3 at the commercial scale and high dose with the rate of 20.2% in leaf TSP and 33.7% in chloroplast TSP. The functional analysis by ELISA confirmed the binding of IMAC purified chloroplast-made DARPin G3 to the extracellular domain of the HER2 receptor with highly effective picomolar affinities. The carcinoma cellular studies by flow cytometry and immunofluorescence microscopy confirmed the correct functioning by the specific binding of the chloroplast-made DARPin G3 to the HER2 receptor on the surface of HER2-positive cancer cell lines.ConclusionThe efficient functional bioactive production of DARPin G3 in chloroplasts led us to introduce plant chloroplasts as the site of efficient production of the first antibody mimetic molecules. This report, as the first case of the cost-effective production of mimetic molecules, enables researchers in pharmaceuticals, synthetic biology, and bio-molecular engineering to develop tool boxes by producing new molecular substitutes for diverse purposes.

RETROSPECTIVE STUDY CORRELATION BETWEEN GRADE AND AMOUNT OF MITOSIS WITH ER, PR, AND HER2 EXPRESSION IN INVASIVE BREAST CARCINOMA OF NO SPECIAL TYPE

Background: In 2018 there were about 24.2% new cases of breast carcinoma worldwide. Molecular breast carcinoma, like expression estrogen receptor (ER), progesterone receptor (PR), and Human Epidermal Growth Factor Receptor2 (HER2) have a higher sensitivity than morphological feature, like tumour grade and mitotic count as prognostic factors. The grade of the tumor reflects how similar these cells are to normal breast epithelial cells. The dominant growth pathway in normal breasts predominantly involves ER and PR, so tumours with ER and PR positivity show lower grade and proliferation activity. In contrast, tumours with ER and PR negativity often have more severe mutation, like overexpression of HER2. Objective: To analyze the correlation between grade and mitotic count with ER, PR, and HER2 expression. Methods: Analytical observational study with a retrospective design. This study used medical record data from ER, PR, and HER2 immunohistochemical examinations as well as histopathological grade and mitotic data for invasive breast carcinoma of no special type in the Anatomical Pathology Laboratory of Dr. Soetomo Surabaya for the period January 1, 2017 - December 31, 2019. Furthermore, the analysis of the relationship between grade and mitotic count was carried out with the expression of ER, PR, and HER2. Results: There was a significant negative relationship between grade and ER with rs = -0.170 (p = 0.000), the number of mitosis and ER with rs = -0.010 (p = 0.001), grade and PR with rs = -0.168 (p = 0.000), and the number of mitosis and PR with rs = -0.110 (p = 0.000). There was no significant relationship between grade and HER2 (p = 0.470) and the number of mitosis and HER2 (p = 0.279). Conclusion: There is a negative significant relationship between grade and amount of mitosis with ER and PR expression. There is no significant relationship between grade and amount of mitosis with HER2 expression.

Review: Radionuclide Molecular Imaging Targeting HER2 in Breast Cancer with a Focus on Molecular Probes into Clinical Trials and Small Peptides.

As the most frequently occurring cancer worldwide, breast cancer (BC) is the leading cause of cancer-related death in women. The overexpression of HER2 (human epidermal growth factor receptor 2) is found in about 15% of BC patients, and it is often associated with a poor prognosis due to the effect on cell proliferation, migration, invasion, and survival. As a result of the heterogeneity of BC, molecular imaging with HER2 probes can non-invasively, in real time, and quantitatively reflect the expression status of HER2 in tumors. This will provide a new approach for patients to choose treatment options and monitor treatment response. Furthermore, radionuclide molecular imaging has the potential of repetitive measurements, and it can help solve the problem of heterogeneous expression and conversion of HER2 status during disease progression or treatment. Different imaging probes of targeting proteins, such as monoclonal antibodies, antibody fragments, nanobodies, and affibodies, are currently in preclinical and clinical development. Moreover, in recent years, HER2-specific peptides have been widely developed for molecular imaging techniques for HER2-positive cancers. This article summarized different types of molecular probes targeting HER2 used in current clinical applications and the developmental trend of some HER2-specific peptides.

Progression of HER-2 lowly expressed breast cancer and the related anti-tumor drugs

Breast cancer is one of the common malignant tumors of women. In recent years, the incidence of breast cancer is high. Human epidermal growth factor receptor-2 (HER-2) is a tyrosine kinase receptor. Breast cancer with abnormal amplification or overexpression of HER-2 have the characteristics of strong tumor invasiveness and poor prognosis. With the advent of anti-HER-2 drugs, the survival period of patients with HER-2 positive breast cancer is gradually prolonged, and the prognosis of patients with HER-2 positive breast cancer is improved. However, the efficacy of traditional HER-2 targeted drugs on patients with low expression of HER-2 is very limited, and the treatment of breast cancer with low expression of HER-2 is still facing challenges. This article reviews the standardization process of the American Society of Clinical Oncology and the American Society of Pathologists guidelines for HER-2 detection, and puts forward the data basis and possibility of defining a new subtype of breast cancer with low expression of HER-2. The birth of a new generation of HER-2 targeting drugs makes it possible to treat patients with low expression of HER-2, which will redefine breast cancer with low expression of HER-2 and provide a new opportunity for the prognosis of patients with low expression of HER-2.

In-silico based identification of phytochemicals from Houttuynia cordata Thunb. as potential inhibitors for overexpressed HER2 and VEGFR2 cancer genes

Human epidermal growth factor receptor2 (HER2) and Vascular endothelial growth factor receptor2 (VEGFR2) – a tyrosine kinase receptors play a key role in breast and stomach cancers. The overexpression of HER2 and VEGFR2 genes increases the number of HER2 and VEGFR2 in the cell which initiates breast and stomach cancer respectively. The phytochemicals from traditional medicinal herb Houttuynia cordata Thunb. are reported to possess anti-inflammatory and anti-cancer potential. However, isolation of phytochemicals from this herb is fraught with uncertainly and time-consuming. Here, a molecular docking approach provides probable binding affinities between the receptors and phytochemicals (ligands) which initiate the first step of anticancer drug discovery and development. In the present study, In-silico docking approaches were used to identify the top-hit phytochemicals from H. cordata as potential inhibitors for overexpressed HER2 (breast) and VEGFR2 (stomach) cancer genes. A total of 100 biologically active phytochemicals from H. cordata were screened and docked against the ligand-binding pocket of HER2 and VEGFR2 kinase domains. Docking results revealed only a few phytochemicals (molecules) which appropriately fit into the ligand-binding pocket with higher binding affinity than the natural ATP ligand. A competitive docking was used to ascertain the top-hit phytochemicals that bind perfectly to the ATP ligand-binding pocket. Among the top-hit phytochemicals docked from H. cordata, the β-sitosterol and Quercetin showed highest binding affinity towards HER2 and VEGFR2 receptors using both hydrogen and hydrophobic interactions. This study confirmed β-sitosterol and Quercetin as potential drug candidates against breast and stomach cancer. Communicated by Ramaswamy H. Sarma

Molecular Mechanism of HER2 Rapid Internalization and Redirected Trafficking Induced by Anti-HER2 Biparatopic Antibody.

Amplification and overexpression of HER2 (human epidermal growth factor receptor 2), an ErbB2 receptor tyrosine kinase, have been implicated in human cancer and metastasis. A bispecific tetravalent anti-HER2 antibody (anti-HER2-Bs), targeting two non-overlapping epitopes on HER2 in domain IV (trastuzumab) and domain II (39S), has been reported to induce rapid internalization and efficient degradation of HER2 receptors. In this study, we investigated the molecular mechanism of this antibody-induced rapid HER2 internalization and intracellular trafficking. Using quantitative fluorescent imaging, we compared the internalization kinetics of anti-HER2-Bs and its parental arm antibodies, alone or in combinations and under various internalization-promoting conditions. The results demonstrated that concurrent engagement of both epitopes was necessary for rapid anti-HER2-Bs internalization. Cellular uptake of anti-HER2-Bs and parental arm antibodies occurred via clathrin-dependent endocytosis; however, inside the cells antibodies directed different trafficking pathways. Trastuzumab dissociated from HER2 in 2 h, enabling the receptor to recycle, whereas anti-HER2-Bs stayed associated with the receptor throughout the entire endocytic pathway, promoting receptor ubiquitination, trafficking to the lysosomes, and efficient degradation. Consistent with routing HER2 to degradation, anti-HER2-Bs significantly reduced HER2 shedding and altered its exosomal export. Collectively, these results enable a better understanding of the mechanism of action of anti-Her2-Bs and can guide the rational design of anti-HER2 therapeutics as well as other bispecific molecules.

Effects of Various Treatment Regimens on the Survival of HER2-positive Breast Cancer Patients with Brain Metastasis

Overexpression of HER2 (human epidermal growth factor receptor 2) occurs in around 20% of breast cancers. This particular subtype is known to be very aggressive and had the worst prognosis prior to the introduction of targeted therapy. The development of anti-HER2 therapies such as trastuzumab and pertuzumab has shown to dramatically improve the prognosis of patients with HER2-positive breast cancer.

HER-2 Protein Overexpression in Patients with Gastric and Oesophageal Adenocarcinoma at a Tertiary Care Facility in Ghana.

The prognosis of gastric and oesophageal adenocarcinoma remains generally poor. However, mounting evidence suggests a positive role of human epidermal growth factor receptor-2 (HER-2) expression in the prognosis of patients with these cancers. In this work, the patterns of HER-2 protein expression were determined in patients with gastric or oesophageal adenocarcinoma. Retrospectively, we reviewed records of gastric and oesophageal biopsies received from 2008 to 2012 and their corresponding archived formalin-fixed paraffin-embedded tissue blocks selected for immunohistochemical analysis. The prevalence of gastric and oesophageal adenocarcinomas and their association with HER-2 protein overexpression were evaluated. Gastric adenocarcinoma made up 18.79% of the gastric biopsies reviewed, and majority of these cancers occurred in males. Regarding the tumour type, HER-2 overexpression was common in the intestinal subtype compared to the diffuse type. Although squamous cell carcinoma was observed to be the commonest (31%) tumour type in the oesophagus compared to adenocarcinoma (8.79%), HER-2 was overexpressed in 42.9% of oesophageal adenocarcinomas, like gastric adenocarcinoma (41.4%). There is a high prevalence of gastric and oesophageal adenocarcinoma, with significant overexpression of HER-2 in these tumours, a window of hope for the management of patients with these cancers.

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.

Tumor characteristics and metabolic quantification in carcinoma breast: An institutional experience.

In India, carcinoma breast is the most common cancer among urban women population and second most common cancer after carcinoma cervix in rural areas. One in 22 women in India develops carcinoma of the breast in their lifetime. Fluorine-18-fluoro-2-deoxy-D-glucose (18F-FDG) uptake in breast cancer usually indicates the degree of tumor metabolism and hence can predict its behavior and prognosis. On the other hand, the estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor-2 (HER-2) or neu state of breast cancer is a biomarker that provides important prognostic information in addition to predicting response to therapy. The main objective of this study is to assess whether a correlation exists between 18F-FDG uptake in untreated cases of breast cancer, their receptor status (ER, PR, and HER-2 or neu), tumor histology, and tumor size. Sixty consecutive female patients, with biopsy-proven primary breast cancer, were enrolled in this prospective study for whom 18F-FDG positron emission tomography-computed tomography scan was done in the Department of Nuclear Medicine. Results obtained were analyzed using appropriate statistical tests (t-test and Pearson Chi-square tests), and interpretation was made with 95% confidence level. In our series, a positive correlation between tumor size, high tumor grade, and standardized uptake value (SUV) was found. Tumors with positive receptor status for estrogen, progesterone, and HER-2/neu receptors had statistically insignificant lower maximum SUV (SUVmax) values than their negative counterparts. Triple-negative breast tumors (ER-, PR-, and no overexpression of HER-2/neu) are currently a subject of major interest because of their aggressiveness, poor prognosis, and lack of targeted therapy. Based on receptor status when the SUVmaxof the group with triple-negative receptor status (ER-/PR-/HER-2/neu-) was compared to rest of the patient group, it was seen that patients with negative receptor status had significantly higher mean SUVmaxvalues. We have inferred that in patients with breast cancer, various biological parameters such as tumor size, grade, histology, and hormonal receptor status have different impact on tumor metabolic activity.

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.

Sensitization of HER2 Positive Breast Cancer Cells to Lapatinib Using Plants-Derived Isothiocyanates

Nearly 25% of all breast cancer is characterized by overexpression of HER2 (human epidermal growth factor receptor 2) which leads to overactivation of prosurvival signal transduction pathways, especially through Akt-mTOR-S6K kinases, and results in enhanced proliferation, migration, induction of angiogenesis, and apoptosis inhibition. Anti-HER2 targeted therapies, such as specific monoclonal antibodies or small-molecule tyrosine kinase inhibitors, even in combination, still seem to be insufficient due to incidence of primary or acquired resistance and prevalence of serious side-effects of these drugs. We assumed that combination of compounds that target different levels of the above-mentioned signal transduction pathway might be more effective in eradication of breast cancer cells. In our in vitro research we used a commercially available drug, lapatinib, acting at the level of the receptor in combination with 1 of the plant-derived isothiocyanates: sulforaphane, erucin, or sulforaphene, as it has been shown previously that sulforaphane inhibits Akt-mTOR-S6K1 pathway in breast cancer cells. We used 2 HER2 overexpressing breast cancer cell lines, SKBR-3 and BT-474. Combinations of the drug and isothiocyanates considerably decreased their viability. This action was synergistic and was accompanied by a decrease in phosphorylation of HER2, Akt, and S6. Combined treatment induced apoptosis more efficiently than either agent alone; however the most effective was a combination of lapatinib with erucin. These findings might support the optimization of therapy based on lapatinib treatment.

Structure–activity relationships of peptidomimetics that inhibit PPI of HER2‐HER3

Human epidermal growth factor receptor-2 (HER2) is a tyrosine kinase family protein receptor that is known to undergo heterodimerization with other members of the family of epidermal growth factor receptors (EGFR) for cell signaling. Overexpression of HER2 and deregulation of signaling has implications in breast, ovarian, and lung cancers. We have designed several peptidomimetics to block the HER2-mediated dimerization, resulting in antiproliferative activity for cancer cells. In this work, we have investigated the structure-activity relationships of peptidomimetic analogs of Compound 5. Compound 5 was conformationally constrained by N- and C-terminal modification and cyclization as well as by substitution with d-amino acids at the N-and C-termini. Among the compounds studied in this work, a peptidomimetic Compound 21 with d-amino acid substitution and its N- and C-termini capped with acetyl and amide functional groups and a reversed sequence compared to that of Compound 5 exhibited better antiproliferative activity in HER2-overexpressed breast, ovarian, and lung cancer cell lines. Compound 21 was further evaluated for its protein-protein interaction (PPI) inhibition ability using enzyme fragment complementation assay, proximity ligation assay, and Western blot analysis. Results suggested that Compound 21 is able to block HER2:HER3 interaction and inhibit phosphorylation of the kinase domain of HER2. The mode of binding of Compound 21 to HER2 protein was modeled using a docking method. Compound 21 seems to bind to domain IV of HER2 near the PPI site of EGFR:HER2, and HER:HER3 and inhibit PPI.

S100A14, a Member of the EF-hand Calcium-binding Proteins, Is Overexpressed in Breast Cancer and Acts as a Modulator of HER2 Signaling

HER2 is overexpressed in 20–25% of breast cancers. Overexpression of HER2 is an adverse prognostic factor and correlates with decreased patient survival. HER2 stimulates breast tumorigenesis via a number of intracellular signaling molecules, including PI3K/AKT and MAPK/ERK.S100A14,one member of the S100 protein family, is significantly associated with outcome of breast cancer patients. Here, for the first time, we show that S100A14 and HER2 are coexpressed in invasive breast cancer specimens,andthere is a significant correlation between the expression levels of the two proteins by immunohistochemistry. S100A14 and HER2 are colocalized in plasma membrane of breast cancer tissue cells and breast cancer cell lines BT474 and SK-BR3. We demonstrate that S100A14 binds directly to HER2 by co-immunoprecipitation and pull-down assays. Further study shows that residues 956–1154 of the HER2 intracellular domain and residue 83 of S100A14 are essential for the two proteins binding.Moreover,we observe a decrease of HER2 phosphorylation, downstream signaling, and HER2-stimulated cell proliferation in S100A14-silenced MCF-7, BT474, and SK-BR3 cells. Our findings suggest that S100A14 functions as a modulator of HER2 signaling and provide mechanistic evidence for its role in breast cancer progression.

Cytoplasmic Overexpression of HER2: A Key Factor in Colorectal Cancer

Trastuzumab, a humanized mouse monoclonal antibody directed against HER2 (Human Epidermal Growth Factor Receptor 2), is currently a keystone in the treatment of breast cancer. Meanwhile, trastuzumab has been validated for use in other types of cancer too. But the data on HER2 expression in colorectal cancer are ambiguous, with reported overexpression of HER2 varying between zero and 84%. In this review these studies are evaluated and compared. It shows that many factors influence the determination of HER2-expression, especially of the intracellular fraction of HER2. It is concluded that although membranous overexpression of HER2 is low in colorectal cancer with only 5% of all patients being positive, a significant proportion of the patients (30%) shows cytoplasmic HER2 overexpression. The clinical impact of enhanced intracellular HER2 is not known, because the nature and origin have not completely been revealed yet. Enlightening this process could be a stepping stone towards targeting of intracellular HER2 as a treatment option.

Abstract 1233: A triad of Brk, HER2 and STAT3 in regulating epithelial and mesenchymal transition and survival of breast cancer cells

Abstract Breast tumor kinase (Brk) is a nonreceptor protein-tyrosine kinase recently shown to be coexpressed with human epidermal growth factor receptor-2 (HER2) at high levels in tumor specimens from patients with invasive ductal breast cancer. Brk is associated with HER2 in breast cancer cells and can directly phosphorylate signal transducers and activators of transcription-3 (STAT3). However, the biological functions of these interactions remain largely unknown. In current study, we developed a subline from HER2-transfected MCF7 breast cancer cells through in vivo selection in nude mice. Overexpression of HER2 led to a higher level of Brk in MCF7 cells and the in vivo selection resulted in a subline with markedly enhanced epithelial and mesenchymal transition (EMT) phenotype and markedly increased migration potential. We named the subline cells EMT1. Both the EMT phenotype and survival of the EMT1 cells heavily depend on Brk, because knockdown of Brk through RNA interference reversed the EMT phenotype, reduced cell migration potential and induced cell death via apoptosis in the EMT1 cells. We further found that the level of activation-specific phosphorylated STAT3 was markedly increased in the EMT1 cells compared with control vector-transfected or HER2-transfected MCF7 cells. Knockdown of Brk significantly reduced the level of STAT3 phosphorylation in the EMT1 cells. Overexpression of a constitutively active STAT3 counteracted Brk knockdown-induced inhibition of cell migration potential and Brk knockdown-induced apoptosis in the EMT1 cells. Together, these preliminary findings support a new paradigm for regulating breast cancer cell EMT and survival that involves interaction and coordination between Brk, HER2, and STAT3. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1233. doi:1538-7445.AM2012-1233

Over-expression of HER-2 is associated with the stage in carcinomas of the urinary bladder

BackgroundThe frequency of over-expression of human epidermal growth factor receptor-2 (HER-2) in bladder cancer is one of the highest among all human malignancies. This over-expression is thought to play a role in aberrant proliferation of cancer cells. Studies on HER-2 expression in bladder carcinoma have shown heterogeneous results.PurposeThe aim of the study was to evaluate the status of HER-2 protein expression in patients with invasive carcinomas of the urinary bladder as related to tumor grade and stage.Materials and methodsArchival samples from 39 patients (6 women, 33 males) with urinary bladder cancer were analyzed for HER-2 over-expression, using immunohistochemistry with the HercepTest.ResultsHER-2 over-expression was observed in 23/39 tumors (59%) and was more frequent in high-grade than in low-grade carcinomas, but the difference was not statistically significant. A significant correlation was established between HER-2 over-expression and tumor stage (p=0.011). HER-2 expression was more frequent in transitional cell carcinomas (TCC) and adenocarcinomas (AC) as compared with squamous cell carcinoma (SCC). Patients’ age and sex were not related to HER-2 over-expression.ConclusionOver-expression of HER-2 was frequent in carcinomas of the urinary bladder. Knowing the HER-2 status would be helpful in formulating a rational treatment strategy for patients with urinary bladder cancer.

Role of HER2 status in the treatment of brain metastases arising from breast cancer by stereotactic radiosurgery.

117 Background: Overexpression of human epidermal growth factor receptor-2 (HER2) in breast cancer has been shown to be an independent risk factor of development of brain metastases (BM). However, the role of HER2 on the radiosensitivity of BM remains controversial. We investigated the efficacy of single fraction stereotactic radiosurgery (SRS) in the treatment of limited BM from breast cancer, based on the HER2 status. Methods: From 2004 to 2010, 57 patients with limited BM were treated with SRS using Leksell Gamma-Knife to a median dose of 20 Gy (range 12-20 Gy) prescribed to the 50% isodose line. The median number of lesions treated were 2 (range 1-7) and the median tumor volume was 92.8 mm3 (range 3.4 -793.7 mm3). There were 27 HER2 positive patients, with 32 ER positive and 22 PR positive patients. Six patients were triple negative. ER/PR information was not available for 4 patients. Overexpression of HER2 was defined as HER2 +++ by immunohistochemistry using the DAKO HercepTest or fluorescent in situ hybridization (FISH) analysis. Time to local recurrence, time to development of new brain metastases, progression-free survival (PFS), and overall survival (OS) were assessed from the date of SRS. Results: With a median follow-up of 11 months (range 1-66 months), 12 patients developed recurrence at the previously treated site (9 HER2 positive patients) and 30 developed new brain metastases (16 HER2 positive patients) The median PFS and OS for all patients were 8 and 16 months respectively. The median PFS for HER2 positive patients compared to HER2 negative patients were 7 and 11 months, respectively (p=0.352). HER2 positive patients had a median OS of 20 months, compared to 14 months in HER2 negative patients (p=0.086). Five patients went on to develop leptomeningeal disease, all of whom were HER2 positive. Conclusions: HER2 overexpression does not increase the risk of recurrence at the site of treatment or appearance of new BM. It does not affect PFS or OS following SRS. However, it remains a risk factor for the development of leptomeningeal disease.

87 BEZ235, a dual PI3K/mTORC inhibitor, targets the DNA damage response leading to radiosensitization and senescence

Growth factors and their receptors play an important role in the pathogenesis of human cancer. The human epidermal growth factor receptor-2 (HER2) is overexpressed in approximately 30% of breast cancers and this is associated with poor clinical outcome. Overexpression of HER2 has been demonstrated to play a direct role in oncogenic transformation. Murine monoclonal antibodies (muMAbs) targeting the extracellular domain of the HER2 receptors suppress HER2-positive cancer cell growth, with muMAb 4D5 having particularly potent activity. A humanized form of muMAb 4D5 was generated by converting all but the antigen-binding region of muMAb 4D5 into human IgG consensus sequences. The humanized monoclonal antibody, Herceptin®, preferentially targets HER2-overexpressing cells, produces responses in breast cancer patients and is well tolerated. In a pivotal phase III trial, Herceptin®administered in combination with chemotherapy (anthracycline/cyclophosphamide or paclitaxel) was compared with chemotherapy alone. The combination was found to produce significant survival benefits in HER2-positive metastatic breast cancer patients. These results have led to the approval of Herceptin®for clinical use in the USA and elsewhere.