Aptasensor with Expanded Nucleotide Using DNA Nanotetrahedra for Electrochemical Detection of Cancerous Exosomes.

Exosomes are extracellular vesicles (50-100 nm) circulating in biofluids as intercellular signal transmitters. Although the potential of cancerous exosomes as tumor biomarkers is promising, sensitive and rapid detection of exosomes remains challenging. Herein, we combined the strengths of advanced aptamer technology, DNA-based nanostructure, and portable electrochemical devices to develop a nanotetrahedron (NTH)-assisted aptasensor for direct capture and detection of hepatocellular exosomes. The oriented immobilization of aptamers significantly improved the accessibility of an artificial nucleobase-containing aptamer to suspended exosomes, and the NTH-assisted aptasensor could detect exosomes with 100-fold higher sensitivity when compared to the single-stranded aptamer-functionalized aptasensor. The present study provides a proof-of-concept for sensitive and efficient quantification of tumor-derived exosomes. We thus expect the NTH-assisted electrochemical aptasensor to become a powerful tool for comprehensive exosome studies.

[1]  Gary K. Schwartz,et al.  Tumour exosome integrins determine organotropic metastasis , 2015, Nature.

[2]  Bob S. Carter,et al.  Chip-based analysis of exosomal mRNA mediating drug resistance in glioblastoma , 2015, Nature Communications.

[3]  David Elashoff,et al.  Electrochemical Sensor for Multiplex Biomarkers Detection , 2009, Clinical Cancer Research.

[4]  Hakho Lee,et al.  Label-free detection and molecular profiling of exosomes with a nano-plasmonic sensor , 2014, Nature Biotechnology.

[5]  Ryan J. White,et al.  An electrochemical supersandwich assay for sensitive and selective DNA detection in complex matrices. , 2010, Journal of the American Chemical Society.

[6]  J. Dear,et al.  Collection, storage, preservation, and normalization of human urinary exosomes for biomarker discovery. , 2006, Kidney international.

[7]  Michael A. Hollingsworth,et al.  Pancreatic cancer exosomes initiate pre-metastatic niche formation in the liver , 2015, Nature Cell Biology.

[8]  H. Nakagawa,et al.  Antibody-coupled monolithic silica microtips for highthroughput molecular profiling of circulating exosomes , 2014, Scientific Reports.

[9]  M. Mascini,et al.  Analytical applications of aptamers. , 2005, Biosensors & bioelectronics.

[10]  Russell P. Goodman,et al.  Rapid Chiral Assembly of Rigid DNA Building Blocks for Molecular Nanofabrication , 2005, Science.

[11]  G. Kristiansen,et al.  Malignant ascites-derived exosomes of ovarian carcinoma patients contain CD24 and EpCAM. , 2007, Gynecologic oncology.

[12]  Hamid Cheshmi Glioblastoma microvesicles transport RNA and proteins that promote tumour growth and provide diagnostic biomarkers , 2011 .

[13]  S. Howorka,et al.  Nanoscale DNA tetrahedra improve biomolecular recognition on patterned surfaces. , 2012, Small.

[14]  Christian Pilarsky,et al.  Glypican-1 identifies cancer exosomes and detects early pancreatic cancer , 2015, Nature.

[15]  S. Pomeroy,et al.  Tumour microvesicles contain retrotransposon elements and amplified oncogene sequences. , 2011, Nature communications.

[16]  George A Calin,et al.  Cancer exosomes perform cell-independent microRNA biogenesis and promote tumorigenesis. , 2014, Cancer cell.

[17]  Kevin W Plaxco,et al.  Rapid, sensitive, and quantitative detection of pathogenic DNA at the point of care through microfluidic electrochemical quantitative loop-mediated isothermal amplification. , 2012, Angewandte Chemie.

[18]  Shana O Kelley,et al.  Interrogating Circulating Microsomes and Exosomes Using Metal Nanoparticles. , 2016, Small.

[19]  Ursula Sauer,et al.  Aptamer-antibody on-chip sandwich immunoassay for detection of CRP in spiked serum. , 2009, Biosensors & bioelectronics.

[20]  Shana O Kelley,et al.  An electrochemical clamp assay for direct, rapid analysis of circulating nucleic acids in serum. , 2015, Nature chemistry.

[21]  S. Campuzano,et al.  Magnetobiosensors based on viral protein p19 for microRNA determination in cancer cells and tissues. , 2014, Angewandte Chemie.

[22]  Graça Raposo,et al.  Extracellular vesicles: Exosomes, microvesicles, and friends , 2013, The Journal of cell biology.

[23]  Paul Harrison,et al.  Classification, Functions, and Clinical Relevance of Extracellular Vesicles , 2012, Pharmacological Reviews.

[24]  A DNA nanostructure for the functional assembly of chemical groups with tunable stoichiometry and defined nanoscale geometry. , 2009, Angewandte Chemie.

[25]  Hao Yan,et al.  A DNA Nanostructure‐based Biomolecular Probe Carrier Platform for Electrochemical Biosensing , 2010, Advanced materials.

[26]  Kevin M. Bradley,et al.  Evolution of functional six-nucleotide DNA. , 2015, Journal of the American Chemical Society.

[27]  Jiye Shi,et al.  Electrochemical detection of nucleic acids, proteins, small molecules and cells using a DNA-nanostructure-based universal biosensing platform , 2016, Nature Protocols.

[28]  B. Sullenger,et al.  Aptamers: an emerging class of therapeutics. , 2005, Annual review of medicine.

[29]  Zhiyuan Hu,et al.  Label-Free Quantitative Detection of Tumor-Derived Exosomes through Surface Plasmon Resonance Imaging , 2014, Analytical chemistry.

[30]  M. Speicher,et al.  Tumor signatures in the blood , 2014, Nature Biotechnology.

[31]  J. Lötvall,et al.  Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells , 2007, Nature Cell Biology.

[32]  Sang-Hee Jeong,et al.  An indirect competitive assay-based aptasensor for detection of oxytetracycline in milk. , 2014, Biosensors & bioelectronics.