HER2 expression plays a key role in various cancers, with overexpression linked to uncontrolled cell growth and metastasis and also its potential as a therapeutic target. Navinci’s advanced in situ proximity ligation technology can provide accurate detection of HER2 and therefore insights into prognosis and treatment decisions. Get your protocol below.
Precise and specific detection of HER2
HER2 expression plays a key role in various cancers, with overexpression linked to uncontrolled cell growth and metastasis and also its potential as a therapeutic target. Navinci’s advanced in situ proximity ligation technology can provide accurate detection of HER2 and therefore insights into prognosis and treatment decisions. Get your protocol below.
In situ detection of HER2 in malignant breast cancer tissue using NaveniBright™ HRP
ErbB/HER receptors in cellular signaling complexity
The human epidermal growth factor receptors (ErbB/HER) consists of four cell surface glycoproteins termed ErbB1/HER1, ErbB2/Neu/HER2, ErbB3/HER3 and ErbB4/HER4 (1,2). ErbB receptors are expressed in a variety of tissues of epithelial, mesenchymal and neuronal origin, where they play fundamental roles in development, proliferation and differentiation via multiple signal transduction. Binding of a ligand to its cognate ErbB receptor induces formation of homo- or heterodimer complexes, which activates receptor tyrosine kinase activity, leading to increased downstream RAS/MAPK, PI3K/AKT, and JAK/STAT signaling (1).
The role of HER2 in diverse cancers
The human epidermal growth factor receptor family of receptors plays a central role in the pathogenesis of several human cancers where they facilitate uncontrolled cell growth and tumorigenesis. Overexpression, amplification, and activating point mutations of Erb/Her family of receptors promote oncogenesis (1). The tumorigenic action of HER2 is not limited to a potential proliferative effect as HER2 has been shown to be a metastasis-promoting factor (3-5).
Most of the studies on HER2 have been carried out in breast cancer after it was found to induce mammary carcinogenesis in vitro and in vivo (6, 7). Overexpression of HER2 is observed in 25–30% of breast cancers and is associated with an aggressive clinical phenotype (1). However, HER2 overexpression also occurs in other forms of cancers such as stomach, ovary, uterine serous endometrial carcinoma, colon, bladder, lung, uterine cervix, head and neck, and esophagus (8,9).
Apart from its role in development of various cancers, it has been intensely evaluated as a therapeutic target. It has also served as a prognostic and predictive biomarker in breast and gastric/gastroesophageal cancers, which have improved the clinical outcome and therapies directed against HER2 have revolutionized the treatment of HER2 overexpressing breast and gastric cancers (5).
In situ detection of HER2 in malignant breast cancer tissue using NaveniBright™ HRP
ErbB/HER receptors in cellular signaling complexity
The human epidermal growth factor receptors (ErbB/HER) consists of four cell surface glycoproteins termed ErbB1/HER1, ErbB2/Neu/HER2, ErbB3/HER3 and ErbB4/HER4 (1,2). ErbB receptors are expressed in a variety of tissues of epithelial, mesenchymal and neuronal origin, where they play fundamental roles in development, proliferation and differentiation via multiple signal transduction. Binding of a ligand to its cognate ErbB receptor induces formation of homo- or heterodimer complexes, which activates receptor tyrosine kinase activity, leading to increased downstream RAS/MAPK, PI3K/AKT, and JAK/STAT signaling (1).
The role of HER2 in diverse cancers
The human epidermal growth factor receptor family of receptors plays a central role in the pathogenesis of several human cancers where they facilitate uncontrolled cell growth and tumorigenesis. Overexpression, amplification, and activating point mutations of Erb/Her family of receptors promote oncogenesis (1). The tumorigenic action of HER2 is not limited to a potential proliferative effect as HER2 has been shown to be a metastasis-promoting factor (3-5).
Most of the studies on HER2 have been carried out in breast cancer after it was found to induce mammary carcinogenesis in vitro and in vivo (6, 7). Overexpression of HER2 is observed in 25–30% of breast cancers and is associated with an aggressive clinical phenotype (1). However, HER2 overexpression also occurs in other forms of cancers such as stomach, ovary, uterine serous endometrial carcinoma, colon, bladder, lung, uterine cervix, head and neck, and esophagus (8,9).
Apart from its role in development of various cancers, it has been intensely evaluated as a therapeutic target. It has also served as a prognostic and predictive biomarker in breast and gastric/ gastroesophageal cancers, which have improved the clinical outcome and therapies directed against HER2 have revolutionized the treatment of HER2 overexpressing breast and gastric cancers (5).
Figure 1. Applying the target-specific Naveni® in situ proximity ligation assay on Her2. Only if the Navenibodies are in close proximity will they generate a rolling circle amplification reaction, leading to a strong and distinct signal
Figure 1. Applying the target-specific Naveni® in situ proximity ligation assay on Her2. Only if the Navenibodies are in close proximity will they generate a rolling circle amplification reaction, leading to a strong and distinct signal
Superior detection of HER2
The two methods currently approved testing for clinical use of HER2 are immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) (10), where the scoring method for HER2 expression is based on the cell membrane staining pattern. We have developed a superior target-specific in situ proximity ligation assay for the visualization of HER2 in tissues. The assay is optimized for single protein detection and features increased sensitivity and efficiency over traditional techniques. Signal will be generated as a result of dual antibody binding to two different epitopes on the same molecule and the subsequent binding of Navenibodies to them, which eliminates off-target effects and greatly improves signal-to-noise ratios compared to standard immunohistochemistry (IHC) (see application examples below). This makes Navincis’ optimized assay for HER2 detection a valuable tool for basic research, as well as an additional verification method in clinical testing with the potential of becoming a new standard.
Superior detection of HER2
The two methods currently approved testing for clinical use of HER2 are immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) (10), where the scoring method for HER2 expression is based on the cell membrane staining pattern. We have developed a superior target-specific in situ proximity ligation assay for the visualization of HER2 in tissues. The assay is optimized for single protein detection and features increased sensitivity and efficiency over traditional techniques. Signal will be generated as a result of dual antibody binding to two different epitopes on the same molecule and the subsequent binding of Navenibodies to them, which eliminates off-target effects and greatly improves signal-to-noise ratios compared to standard immunohistochemistry (IHC) (see application examples below). This makes Navincis’ optimized assay for HER2 detection a valuable tool for basic research, as well as an additional verification method in clinical testing with the potential of becoming a new standard.
Application example – Detection of HER2, comparison Naveni technology vs IHC
Accurate detection of HER2 is essential. The application examples below compare the Naveni technology, utilizing a dual antibody recognition approach, with traditional IHC based on single antibodies.
Application example – Detection of HER2, comparison Naveni technology vs IHC
Accurate detection of HER2 is essential. The application examples below compare the Naveni technology, utilizing a dual antibody recognition approach, with traditional IHC based on single antibodies.
In situ proximity ligation assay detection of HER2 in breast tissue, showing positive staining in malignant tissue. Signals are visualised using NaveniBright
In situ proximity ligation assay detection of HER2 in breast tissue, showing negative staining in healthy tissue. Signals are visualised using NaveniBright
In situ proximity ligation assay detection of HER2 in breast tissue, image one shows positive staining in malignant tissue and image two negative staining in healthy tissue. Signals are visualised using NaveniBright HRP
IHC detection of HER2 in breast tissue, showing positive staining in malignant tissue, A using rabbit mAb and B using mouse mAb
IHC detection of HER2 in breast tissue, showing unspecific staining of HER2 given that the breast tissue is not expressing HER2
IHC detection of HER2 in breast tissue, image one shows positive staining in malignant tissue, A using rabbit mAb and B using mouse mAb. Image two shows unspecific staining of HER2 given that the breast tissue is not expressing HER2
How to detect HER2 with precision
Use our flexible products NaveniBright HRP or NaveniFlex Tissue. For further information about HER2 monoclonal antibodies and protocols, please contact us using the form below.
How to detect HER2 with precision
Use our flexible products NaveniBright HRP or NaveniFlex Tissue. For further information about HER2 monoclonal antibodies and protocols, please contact us using the form below.
NaveniBright HRP
The assay is designed to be used with a mouse and a rabbit primary pair. Chromogenic readout with HRP substrate.
NaveniFlex Tissue
The assay is designed to be used with a mouse and rabbit primary antibody pair. Detection with Atto647N fluorophore is included.
NaveniBright HRP
The assay is designed to be used with a mouse and a rabbit primary pair. Chromogenic readout with HRP substrate.
NaveniFlex Tissue
The assay is designed to be used with a mouse and rabbit primary antibody pair. Detection with Atto647N fluorophore is included.
References
- Olayioye, M. A., Neve, R. M., Lane, H. A., & Hynes, N. E. (2000). The ErbB signaling network: receptor heterodimerization in development and cancer. The EMBO journal, 19(13), 3159–3167. DOI: 10.1093/emboj/19.13.3159
- Roskoski R., Jr (2019). Small molecule inhibitors targeting the EGFR/ErbB family of protein-tyrosine kinases in human cancers. Pharmacological research, 139, 395–411. DOI: 10.1016/j.phrs.2018.11.014
- Olayioye M. A. (2001). Update on HER-2 as a target for cancer therapy: intracellular signaling pathways of ErbB2/HER-2 and family members. Breast cancer research : BCR, 3(6), 385–389. DOI: 10.1186/bcr327
- Neve, R. M., Lane, H. A., & Hynes, N. E. (2001). The role of overexpressed HER2 in transformation. Annals of Oncology, 12, S9–S13. DOI: 10.1093/annonc/12.suppl_1.S9
- Ménard, S., Pupa, S. M., Campiglio, M., & Tagliabue, E. (2003). Biologic and therapeutic role of HER2 in cancer. Oncogene, 22(42), 6570–6578. DOI: 10.1038/sj.onc.1206779
- Di Fiore, P. P., Pierce, J. H., Kraus, M. H., Segatto, O., King, C. R., & Aaronson, S. A. (1987). Erb B-2 Is a Potent Oncogene When Overexpressed in NIH/3T3 Cells. Science, 237(4811), 178–182. DOI: 10.1126/science.2885917
- Muller, W. J., Sinn, E., Pattengale, P. K., Wallace, R., & Leder, P. (1988). Single-step induction of mammary adenocarcinoma in transgenic mice bearing the activated c-neu oncogene. Cell, 54(1), 105–115. DOI: 10.1016/0092-8674(88)90184-5
- Fukushige, S., Matsubara, K., Yoshida, M., Sasaki, M., Suzuki, T., Semba, K., Toyoshima, K., & Yamamoto, T. (1986). Localization of a novel v-erbB-related gene, c-erbB-2, on human chromosome 17 and its amplification in a gastric cancer cell line. Molecular and Cellular Biology, 6(3), 955–958. DOI: 10.1128/MCB.6.3.955
- Reichelt, U., Duesedau, P., Tsourlakis, M. C., Quaas, A., Link, B. C., Schurr, P. G., Kaifi, J. T., Gros, S. J., Yekebas, E. F., Marx, A., Simon, R., Izbicki, J. R., & Sauter, G. (2007). Frequent homogeneous HER-2 amplification in primary and metastatic adenocarcinoma of the esophagus. Modern Pathology, 20(1), 120–129. https://doi.org/10.1038/modpathol.3800712
- Wolff, A. C., Hammond, M. E. H., Hicks, D. G., Dowsett, M., McShane, L. M., Allison, K. H., Allred, D. C., Bartlett, J. M. S., Bilous, M., Fitzgibbons, P., Hanna, W., Jenkins, R. B., Mangu, P. B., Paik, S., Perez, E. A., Press, M. F., Spears, P. A., Vance, G. H., Viale, G., & Hayes, D. F. (2013). Recommendations for Human Epidermal Growth Factor Receptor 2 Testing in Breast Cancer: American Society of Clinical Oncology/College of American Pathologists Clinical Practice Guideline Update. Journal of Clinical Oncology, 31(31), 3997–4013. DOI: 10.1200/JCO.2013.50.9984