Selection of DNA aptamers for extra cellular domain of human epidermal growth factor receptor 2 to detect HER2 positive carcinomas

Sett, Arghya; Borthakur, Bibhuti Bhusan; Bora, Utpal

doi:10.1007/s12094-017-1629-y

Cited by 33 publications

(17 citation statements)

References 29 publications

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“…As shown in Figure 3A, after the fixed SKBR-3 cells were treated with Apt2-G4 probes and implemented intact DCTDR system, cells showed bright and clear green fluorescence, indicating the Apt2-G4 probes can bind with the HER2 proteins on the cell surface and triggered DCTDR in situ for HER2 protein imaging. The fluorescence signals distributed mainly on the cell surface but partly in cytoplasmic surroundings of SKBR-3 cells, which is in accordance with the imaging results of the original article for this aptamer development, due to enhanced cell penetration ability of DNA aptamer 23. Meanwhile, there was no detectable fluorescence image, when Apt2-G4 or hemin was absent in the DCTDR system.…”

Section: Resultssupporting

confidence: 85%

“…HER2 protein was chosen as the model. HER2 specific aptamer (Apt2) 23 and intact G-quadruplex sequence (G4) were incorporated to design a bifunctional Apt2-G4 probes for total HER2 protein analysis. The recognition of HER2 protein with the aptamer part can present the hemin/G4 DNAzyme on the cell surface and subsequently initiated DCTDR in situ.…”

Section: Introductionmentioning

confidence: 99%

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DNAzyme Catalyzed Tyramide Depositing Reaction for In Situ Imaging of Protein Status on the Cell Surface

Xu¹,

Liu²,

Yang³

et al. 2019

Theranostics

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Effective characterization of protein biomarkers status on the cell surface has important value in the diagnosis and treatment of diseases. Traditional immunohistochemistry can only assess the protein expression level rather than accurately reflect their interaction and oligomerization, resulting in inevitable problems for personalized therapy. Methods: Herein, we developed a novel DNAzyme-catalyzed tyramide depositing reaction (DCTDR) for in situ amplified imaging of membrane protein status. By using human epidermal growth factor receptor 2 (HER2) as model, the binding of HER2 proteins with specific aptamers induced the formation of activated hemin/G-quadruplex (G4) DNAzyme on the cell surface to catalyze the covalent deposition of fluorescent tyramide on the membrane proteins for fluorescence imaging. Results: The DCTDR-based imaging can conveniently characterize total HER2 expression and HER2 dimerization on the breast cancer cell surface with the application of aptamer-G4 probes and proximity aptamer-split G4 probes, respectively. The designed DCTDR strategy was successfully applied to quantitatively estimate total HER2 expression and HER2 homodimer on clinical breast cancer tissue sections with high specificity and accuracy. Conclusion: The DCTDR strategy provides a simple, pragmatic and enzyme-free toolbox to conveniently and sensitively analyze protein status in clinical samples for improving clinical research, cancer diagnostics and personalized treatment.

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Section: Resultssupporting

confidence: 85%

Section: Introductionmentioning

confidence: 99%

DNAzyme Catalyzed Tyramide Depositing Reaction for In Situ Imaging of Protein Status on the Cell Surface

Xu¹,

Liu²,

Yang³

et al. 2019

Theranostics

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show abstract

“…Apart from this, several aptamers were also developed to target human epidermal growth factor 2. These were used to deliver therapeutic drugs, RNA, and nanoparticles into the cancer cells [ 76 , 77 ].…”

Section: Aptamers As Therapeutic Agentsmentioning

confidence: 99%

Current Perspectives on Aptamers as Diagnostic Tools and Therapeutic Agents

2020

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Aptamers are synthetic single-stranded DNA or RNA sequences selected from combinatorial oligonucleotide libraries through the well-known in vitro selection and iteration process, SELEX. The last three decades have witnessed a sudden boom in aptamer research, owing to their unique characteristics, like high specificity and binding affinity, low immunogenicity and toxicity, and ease in synthesis with negligible batch-to-batch variation. Aptamers can specifically bind to the targets ranging from small molecules to complex structures, making them suitable for a myriad of diagnostic and therapeutic applications. In analytical scenarios, aptamers are used as molecular probes instead of antibodies. They have the potential in the detection of biomarkers, microorganisms, viral agents, environmental pollutants, or pathogens. For therapeutic purposes, aptamers can be further engineered with chemical stabilization and modification techniques, thus expanding their serum half-life and shelf life. A vast number of antagonistic aptamers or aptamer-based conjugates have been discovered so far through the in vitro selection procedure. However, the aptamers face several challenges for its successful clinical translation, and only particular aptamers have reached the marketplace so far. Aptamer research is still in a growing stage, and a deeper understanding of nucleic acid chemistry, target interaction, tissue distribution, and pharmacokinetics is required. In this review, we discussed aptamers in the current diagnostics and theranostics applications, while addressing the challenges associated with them. The report also sheds light on the implementation of aptamer conjugates for diagnostic purposes and, finally, the therapeutic aptamers under clinical investigation, challenges therein, and their future directions.

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“…This RNA aptamer could bind with HER2 positive cell line MDA-MB-453 and KPL-4 but had no affinity towards HER2 negative cells such as MCF-7 and A431. In another study, Sett et al reported the isolation of DNA aptamer ECD_Apt1 to specifically target extracellular domain of HER2 protein [ 67 ] and then conjugated the ECD_Apt1 aptamer with biotin. This biotin-aptamer conjugate showed stronger cytoplasmic staining in SKBR3 compared to MDA-MB-231 and MCF-7.…”

Section: Applications Of Aptamer In Breast Cancermentioning

confidence: 99%

Potential Diagnostic and Therapeutic Applications of Oligonucleotide Aptamers in Breast Cancer

Shaikh

et al. 2017

IJMS

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Breast cancer is one of the most common causes of cancer related deaths in women. Currently, with the development of early detection, increased social awareness and kinds of treatment options, survival rate has improved in nearly every type of breast cancer patients. However, about one third patients still have increased chances of recurrence within five years and the five-year relative survival rate in patients with metastasis is less than 30%. Breast cancer contains multiple subtypes. Each subtype could cause distinct clinical outcomes and systemic interventions. Thereby, new targeted therapies are of particular importance to solve this major clinical problem. Aptamers, often termed “chemical antibodies”, are functionally similar to antibodies and have demonstrated their superiority of recognizing target with high selectivity, affinity and stability. With these intrinsic properties, aptamers have been widely studied in cancer biology and some are in clinical trials. In this review, we will firstly discuss about the global impacts and mechanisms of breast cancer, then briefly highlight applications of aptamers that have been developed for breast cancer and finally summarize various challenges in clinical translation of aptamers.

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Selection of DNA aptamers for extra cellular domain of human epidermal growth factor receptor 2 to detect HER2 positive carcinomas

Cited by 33 publications

References 29 publications

DNAzyme Catalyzed Tyramide Depositing Reaction for In Situ Imaging of Protein Status on the Cell Surface

DNAzyme Catalyzed Tyramide Depositing Reaction for In Situ Imaging of Protein Status on the Cell Surface

Current Perspectives on Aptamers as Diagnostic Tools and Therapeutic Agents

Potential Diagnostic and Therapeutic Applications of Oligonucleotide Aptamers in Breast Cancer

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