Sensitive fluorescent detection of exosomal microRNA based on enzymes-assisted dual-signal amplification

Xia, Yaokun; Huang, Ze-Ning; Chen, Tingting; Xu, Lilan; Zhu, Gengzhen; Chen, Wenqian; Chen, Guan-Yu; Wu, Shu-Xiang; Lan, Jianming; Lin, Xu; Chen, Jinghua

doi:10.1016/j.bios.2022.114259

Cited by 11 publications

(8 citation statements)

References 54 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In our study, the widely used biomarker of EVs miR-21 was chosen. − miR-21, one of the mammalian miRNAs discovered, is located in the 11th intron of the TMEM49 (transmembrane protein 49) gene on chromosome 17 (17q.23.1), which is the precursor of vacuole membrane protein 1 (VMP1). miR-21 increases the cell proliferation and decreases apoptosis, promoting the incidence of various cancers by activating oncogenic signaling pathways.…”

Section: Resultsmentioning

confidence: 99%

Localized Imaging of Programmed Death-Ligand 1 on Individual Tumor-Derived Extracellular Vesicles for Prediction of Immunotherapy Response

Zhang,

Guan,

et al. 2023

ACS Nano

View full text Add to dashboard Cite

Programmed death-ligand 1 (PD-L1) on tumorderived small extracellular vesicles (EVs) is a biomarker for prediction of the immunotherapy response. However, conventional bulk measurement can hardly analyze the expression of PD-L1 on individual tumor-derived EVs. Herein, a method for localized imaging of tumor-derived individual EVs PD-L1 (LITIE) is developed. In this assay, EVs in plasma were directly captured on a biochip. Then the liposome-mediated membrane fusion strategy was used to image miR-21 in EVs to discriminate miR-21-positive EVs from the whole EVs populations. Subsequently, the primer exchange reaction (PER) is applied to generate localized and amplified fluorescent signals for imaging PD-L1 on identified tumor-derived EVs. When applied in clinical sample tests, the LITIE assay could effectively distinguish breast cancer patients from healthy donors or patients with benign tumors. Interestingly, in a mice melanoma model, the LITIE assay showed the ability to predict immunotherapy response even before drug treatment. Thus, we think the strategy of measuring individual tumor-derived EVs PD-L1 could serve as an alternative way for screening clinical responders suitable for immunotherapy.

show abstract

Section: Resultsmentioning

confidence: 99%

Localized Imaging of Programmed Death-Ligand 1 on Individual Tumor-Derived Extracellular Vesicles for Prediction of Immunotherapy Response

Zhang,

Guan,

et al. 2023

ACS Nano

View full text Add to dashboard Cite

show abstract

“…By detecting EV miR-21, GC patients can be distinguished from PC lesions patients with an accuracy of 83.9%. [50] RCA is a commonly used enzyme-assisted isothermal amplification method. RCA uses a small segment of cyclic oligonucleotide as a template and deoxynucleoside triphosphates or nucleoside triphosphate as raw material to generate a long repetitive single-stranded DNA or RNA by amplification under the action of DNA or RNA polymerase.…”

Section: Enzyme Signal Amplificationmentioning

confidence: 99%

“…By detecting EV miR‐21, GC patients can be distinguished from PC lesions patients with an accuracy of 83.9%. [ 50 ]…”

Section: Fluorescent Detectionmentioning

confidence: 99%

Emerging Sensing and In Situ Detection Technologies for the Analysis of Extracellular Vesicle miRNAs

Han,

Wang,

Zhu

et al. 2023

Advanced NanoBiomed Research

View full text Add to dashboard Cite

Liquid biopsy has received increasing attention as a new disease detection modality because of its noninvasive, simple sampling, and reproducible assay advantages. Among the markers of liquid biopsy, extracellular vesicles (EVs) are considered as promising disease biomarkers because they contain a large amount of biological information and have a significant role in physiological activities. The emergence and progression of some of these diseases are associated with miRNAs carried by EVs (EV‐miRNAs). Therefore, high‐sensitive detection of EV‐miRNAs is essential in clinical applications. A growing number of strategies, including biosensors, in situ detection methods, and microfluidics have been developed for the detection of EV‐miRNA and have been applied in the diagnosis of diseases such as cancer. This review summarizes the probes, signal amplification, and detection methods for EV‐miRNA detection, as well as the application of membrane fusion‐based in situ detection and integrated microfluidic chips for EV‐miRNA detection. The challenges of these materials and techniques in clinical diagnostic applications are also discussed.

show abstract

“…Owing to high programmability and biocompatibility, DNA has shown great potential as a building block for engineering a nanosensor that can detect a variety of targets, including nucleic acid, proteins, and metal ions. − In particular, a wide range of DNA probes modified with different functional groups have been designed for the individual detection of GSH or APE1 in test tubes or living cells. − In spite of the success acquired in this area, the predictable control over the target molecular imaging at the right site was still limited, particularly in living organisms. Recently, efforts have focused on imparting stimuli-responsive capabilities to DNA-based sensors to allow conditional control over its functions.…”

Section: Introductionmentioning

confidence: 99%

AND-Gated Nanosensor for Imaging of Glutathione and Apyrimidinic Endonuclease 1 in Cells, Animals, and Organoids

Huang

Zhao

et al. 2023

ACS Appl. Mater. Interfaces

Self Cite

View full text Add to dashboard Cite

The development of a strategy for imaging of glutathione (GSH) and apurinic/apyrimidinic endonuclease 1 (APE1) in an organism remains challenging despite their significance in elaborating the correlated pathophysiological processes. Therefore, in this study, we propose a DNA-based AND-gated nanosensor for fluorescence imaging of the GSH as well as APE1 in living cells, animals, and organoids. The DNA probe is composed of a G-strand and A-strand. The disulfide bond in the G-strand is cleaved through a GSH redox reaction, and the hybridization stability between the G-strand and A-strand is decreased, leading to a conformational change of the A-strand. In the presence of APE1, the apurinic/apyrimidinic (AP) site in the A-strand is digested, producing a fluorescence signal for the correlated imaging of GSH and APE1. This nanosensor enables monitoring of the expression level change of GSH and APE1 in cells. Additionally, we illustrate the capability of this “dual-keys-and-locked” conceptual methodology in achieving specific tumor imaging when GSH and APE1 are present simultaneously (overexpressed GSH and APE1 in tumor cells) with improving tumor-to-normal tissue ratio in vivo. Furthermore, using this nanosensor, the GSH and APE1 also are visualized in organoids that recapitulate the phenotypic and functional traits of the original biological specimens. Overall, this study demonstrates the potential of our proposed biosensing technology in investigating the roles of various biological molecules involved in specific diseases.

show abstract

Sensitive fluorescent detection of exosomal microRNA based on enzymes-assisted dual-signal amplification

Cited by 11 publications

References 54 publications

Localized Imaging of Programmed Death-Ligand 1 on Individual Tumor-Derived Extracellular Vesicles for Prediction of Immunotherapy Response

Localized Imaging of Programmed Death-Ligand 1 on Individual Tumor-Derived Extracellular Vesicles for Prediction of Immunotherapy Response

Emerging Sensing and In Situ Detection Technologies for the Analysis of Extracellular Vesicle miRNAs

AND-Gated Nanosensor for Imaging of Glutathione and Apyrimidinic Endonuclease 1 in Cells, Animals, and Organoids

Contact Info

Product

Resources

About