Amplified fluorescence imaging of HER2 dimerization on cancer cells by using a co-localization triggered DNA nanoassembly
- 28 Downloads
Convenient and sensitive detection of human epidermal growth factor receptor 2 (HER2) dimerization is highly desirable for molecule subtyping and guiding personalized HER2 targeted therapy of breast cancer. A colocalization-triggered DNA nanoassembly (CtDNA) strategy was developed for amplified imaging of HER2 dimerization. It exploits (a) the advantage of the specificity of aptamer proximity hybridization, and (b) the high sensitivity of hairpin-free nonlinear HCR. The mechanism of step-by-step hairpin-free nonlinear HCR for DNA dendritic nanoassembly was studied by native polyacrylamide gel electrophoresis, atomic force microscopy and fluorometry. The results revealed a high specificity, sensitivity, and excellent controllability of the DNA dendritic nanoassembly. The method was used to identify HER2 homodimers and HER2/HER3 heterodimers in various breast cancer cell lines using fluorescence microscopy. It was then extended to image and quantitatively evaluate HER2 homodimers in clinical formalin-fixed paraffin-embedded breast cancer tissue specimens. This revealed its remarkable accuracy and practicality for clinical diagnostics.
KeywordsHER2 dimerization Fluorescent imaging Aptamer Proximity hybridization Hairpin-free nonlinear HCR DNA nanoassembly Formalin-fixed paraffin-embedded tissue Breast cancer Cancer diagnostics
This work was funded by the National Natural Science Foundation of China (81572080, 81873972 and 81873980) and the Training Program for Advanced Young Medical Personnel of Chongqing (2017HBRC003).
Compliance with ethical standards
The author(s) declare that they have no competing interests.
- 8.Yoshioka Y, Suzuki T, Matsuo Y, Tsurita G, Watanabe T, Dohmae N, Nakamura Y, Hamamoto R (2017) Protein lysine methyltransferase SMYD3 is involved in tumorigenesis through regulation of HER2 homodimerization. Cancer med 6(7):1665–1672. https://doi.org/10.1002/cam4.1099 CrossRefPubMedPubMedCentralGoogle Scholar
- 10.Ghosh R, Narasanna A, Wang SE, Liu S, Chakrabarty A, Balko JM, Gonzalez-Angulo AM, Mills GB, Penuel E, Winslow J, Sperinde J, Dua R, Pidaparthi S, Mukherjee A, Leitzel K, Kostler WJ, Lipton A, Bates M, Arteaga CL (2011) Trastuzumab has preferential activity against breast cancers driven by HER2 homodimers. Cancer Res 71(5):1871–1882. https://doi.org/10.1158/0008-5472.can-10-1872 CrossRefPubMedPubMedCentralGoogle Scholar
- 12.Spears M, Taylor KJ, Munro AF, Cunningham CA, Mallon EA, Twelves CJ, Cameron DA, Thomas J, Bartlett JM (2012) In situ detection of HER2:HER2 and HER2:HER3 protein-protein interactions demonstrates prognostic significance in early breast cancer. Breast Cancer Res Treat 132(2):463–470. https://doi.org/10.1007/s10549-011-1606-z CrossRefPubMedGoogle Scholar
- 16.Wang L, Li W, Sun J, Zhang S-Y, Yang S, Li J, Li J, Yang H-H (2018) Imaging of receptor dimers in zebrafish and living cells via aptamer recognition and proximity-induced hybridization chain reaction. Anal Chem 90(24):14433–14438. https://doi.org/10.1021/acs.analchem.8b04015 CrossRefPubMedGoogle Scholar
- 23.Chang CC, Chen CY, Chuang TL, Wu TH, Wei SC, Liao H, Lin CW (2016) Aptamer-based colorimetric detection of proteins using a branched DNA cascade amplification strategy and unmodified gold nanoparticles. Biosens Bioelectron 78:200–205. https://doi.org/10.1016/j.bios.2015.11.051 CrossRefPubMedGoogle Scholar
- 26.Ding X, Cheng W, Li Y, Wu J, Li X, Cheng Q, Ding S (2017) An enzyme-free surface plasmon resonance biosensing strategy for detection of DNA and small molecule based on nonlinear hybridization chain reaction. Biosens Bioelectron 87:345–351. https://doi.org/10.1016/j.bios.2016.08.077 CrossRefPubMedGoogle Scholar
- 27.Chen CH, Chernis GA, Hoang VQ, Landgraf R (2003) Inhibition of heregulin signaling by an aptamer that preferentially binds to the oligomeric form of human epidermal growth factor receptor-3. Proc Natl Acad Sci U S A 100(16):9226–9231. https://doi.org/10.1073/pnas.1332660100 CrossRefPubMedPubMedCentralGoogle Scholar
- 28.Shi Y, Huang W, Tan Y, Jin X, Dua R, Penuel E, Mukherjee A, Sperinde J, Pannu H, Chenna A, DeFazio-Eli L, Pidaparthi S, Badal Y, Wallweber G, Chen L, Williams S, Tahir H, Larson J, Goodman L, Whitcomb J, Petropoulos C, Winslow J (2009) A novel proximity assay for the detection of proteins and protein complexes: quantitation of HER1 and HER2 total protein expression and homodimerization in formalin-fixed, paraffin-embedded cell lines and breast cancer tissue. Diagn Mol Pathol 18(1):11–21. https://doi.org/10.1097/PDM.0b013e31818cbdb2 CrossRefPubMedGoogle Scholar
- 29.Garner AP, Bialucha CU, Sprague ER, Garrett JT, Sheng Q, Li S, Sineshchekova O, Saxena P, Sutton CR, Chen D, Chen Y, Wang H, Liang J, Das R, Mosher R, Gu J, Huang A, Haubst N, Zehetmeier C, Haberl M, Elis W, Kunz C, Heidt AB, Herlihy K, Murtie J, Schuller A, Arteaga CL, Sellers WR, Ettenberg SA (2013) An antibody that locks HER3 in the inactive conformation inhibits tumor growth driven by HER2 or neuregulin. Cancer Res 73(19):6024–6035. https://doi.org/10.1158/0008-5472.can-13-1198 CrossRefPubMedPubMedCentralGoogle Scholar
- 30.Okita R, Mougiakakos D, Ando T, Mao Y, Sarhan D, Wennerberg E, Seliger B, Lundqvist A, Mimura K, Kiessling R (2012) HER2/HER3 signaling regulates NK cell-mediated cytotoxicity via MHC class I chain-related molecule a and B expression in human breast cancer cell lines. J Immunol 188(5):2136–2145. https://doi.org/10.4049/jimmunol.1102237 CrossRefPubMedGoogle Scholar
- 32.Wolff AC, Hammond MEH, Allison KH, Harvey BE, Mangu PB, Bartlett JMS, Bilous M, Ellis IO, Fitzgibbons P, Hanna W, Jenkins RB, Press MF, Spears PA, Vance GH, Viale G, McShane LM, Dowsett M (2018) Human epidermal growth factor receptor 2 testing in breast Cancer: American Society of Clinical Oncology/College of American Pathologists Clinical Practice Guideline Focused Update. Arch Pathol Lab Med 36(20):2105–2122. https://doi.org/10.1200/jco.2018.77.8738 CrossRefGoogle Scholar
- 33.Huang W, Reinholz M, Weidler J, Yolanda L, Paquet A, Whitcomb J, Lingle W, Jenkins RB, Chen B, Larson JS, Tan Y, Sherwood T, Bates M, Perez EA (2010) Comparison of central HER2 testing with quantitative total HER2 expression and HER2 homodimer measurements using a novel proximity-based assay. Am J Clin Pathol 134(2):303–311. https://doi.org/10.1309/ajcp3bzy4yafntrg CrossRefPubMedGoogle Scholar