Aptamer/AuNP Biosensor for Colorimetric Profiling of Exosomal Proteins.

Exosomes constitute an emerging biomarker for cancer diagnosis because they carry multiple proteins that reflect the origins of parent cells. Assessing exosome surface proteins provides a powerful means of identifying a combination of biomarkers for cancer diagnosis. We report a sensor platform that profiles exosome surface proteins in minutes by the naked eye. The sensor consists of a gold nanoparticle (AuNP) complexed with a panel of aptamers. The complexation of aptamers with AuNPs protects the nanoparticles from aggregating in a high-salt solution. In the presence of exosomes, the non-specific and weaker binding between aptamers and the AuNP is broken, and the specific and stronger binding between exosome surface protein and the aptamer displaces aptamers from the AuNP surface and results in AuNP aggregation. This aggregation results in a color change and generates patterns for the identification of multiple proteins on the exosome surface.

[1]  Wifredo Ricart,et al.  The version of record : , 2018 .

[2]  Joshua E. Smith,et al.  Gold nanoparticle-based colorimetric assay for the direct detection of cancerous cells. , 2008, Analytical chemistry.

[3]  P. Park,et al.  Discovering statistically significant pathways in expression profiling studies. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[4]  M. V. Filatov,et al.  Exosomes are natural carriers of exogenous siRNA to human cells in vitro , 2013, Cell Communication and Signaling.

[5]  D. Astruc,et al.  Gold nanoparticles: assembly, supramolecular chemistry, quantum-size-related properties, and applications toward biology, catalysis, and nanotechnology. , 2004, Chemical reviews.

[6]  R. Simpson,et al.  Proteomic profiling of exosomes: Current perspectives , 2008, Proteomics.

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

[8]  John J Rossi,et al.  Cell-specific aptamer-mediated targeted drug delivery. , 2011, Oligonucleotides.

[9]  Laurence Zitvogel,et al.  Exosomes: composition, biogenesis and function , 2002, Nature Reviews Immunology.

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

[11]  Huixiang Li,et al.  Colorimetric detection of DNA sequences based on electrostatic interactions with unmodified gold nanoparticles. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[12]  C. Théry,et al.  Membrane vesicles as conveyors of immune responses , 2009, Nature Reviews Immunology.

[13]  Yusuke Yoshioka,et al.  Ultra-sensitive liquid biopsy of circulating extracellular vesicles using ExoScreen , 2014, Nature Communications.

[14]  J Christopher Love,et al.  Cell-surface sensors for real-time probing of cellular environments. , 2011, Nature nanotechnology.

[15]  Nan Zhang,et al.  DNA Aptamer Evolved by Cell-SELEX for Recognition of Prostate Cancer , 2014, PloS one.

[16]  R. Schiffelers,et al.  Exosome mimetics: a novel class of drug delivery systems , 2012, International journal of nanomedicine.

[17]  L. Gold,et al.  Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. , 1990, Science.

[18]  S. Q. Xie,et al.  Proteomic Analysis of Mitotic RNA Polymerase II Reveals Novel Interactors and Association With Proteins Dysfunctional in Disease* , 2011, Molecular & Cellular Proteomics.

[19]  Tushar Patel,et al.  Development of an aptasensor for electrochemical detection of exosomes. , 2016, Methods.

[20]  Cuichen Wu,et al.  Engineering of switchable aptamer micelle flares for molecular imaging in living cells. , 2013, ACS nano.

[21]  J. Serrano,et al.  Orthogonal action of noncovalent interactions for photoresponsive chiral columnar assemblies. , 2010, Angewandte Chemie.

[22]  Hakho Lee,et al.  Protein typing of circulating microvesicles allows real-time monitoring of glioblastoma therapy , 2012, Nature Medicine.

[23]  R. Setterquist,et al.  Exosomes: current knowledge of their composition, biological functions, and diagnostic and therapeutic potentials. , 2012, Biochimica et biophysica acta.

[24]  J. Rossi,et al.  Aptamers as targeted therapeutics: current potential and challenges , 2016, Nature Reviews Drug Discovery.

[25]  Yuexiang Lu,et al.  Aptamer-based plasmonic sensor array for discrimination of proteins and cells with the naked eye. , 2013, Analytical chemistry.

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

[27]  W. Tan,et al.  Molecular Recognition-Based DNA Nanoassemblies on the Surfaces of Nanosized Exosomes. , 2017, Journal of the American Chemical Society.

[28]  Weihong Tan,et al.  Cell-specific aptamer probes for membrane protein elucidation in cancer cells. , 2008, Journal of proteome research.

[29]  Salma Khan,et al.  Survivin is released from cancer cells via exosomes , 2010, Apoptosis.

[30]  Rong-Fong Shen,et al.  Identification and proteomic profiling of exosomes in human urine. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[31]  S. Lim,et al.  Hypoxic Tumor Cell Modulates Its Microenvironment to Enhance Angiogenic and Metastatic Potential by Secretion of Proteins and Exosomes* , 2010, Molecular & Cellular Proteomics.

[32]  W. Duan,et al.  Selection of DNA aptamers against epithelial cell adhesion molecule for cancer cell imaging and circulating tumor cell capture. , 2013, Analytical chemistry.

[33]  H. Adomat,et al.  Exosomes as Biomarker Enriched Microvesicles: Characterization of Exosomal Proteins Derived from a Panel of Prostate Cell Lines with Distinct AR Phenotypes , 2012, Molecular & Cellular Proteomics.

[34]  Weihong Tan,et al.  Cell-specific internalization study of an aptamer from whole cell selection. , 2008, Chemistry.

[35]  Ronghua Yang,et al.  Functional DNA-Containing Nanomaterials: Cellular Applications in Biosensing, Imaging, and Targeted Therapy , 2014, Accounts of chemical research.

[36]  Chad A Mirkin,et al.  Nanostructures in biodiagnostics. , 2005, Chemical reviews.

[37]  Steven A Greenberg,et al.  A gene expression approach to study perturbed pathways in myositis , 2007, Current opinion in rheumatology.

[38]  Weihong Tan,et al.  Synthetic DNA Aptamers to Detect Protein Molecular Variants in a High‐Throughput Fluorescence Quenching Assay , 2003, Chembiochem : a European journal of chemical biology.

[39]  Ping Yu,et al.  A simple assay for direct colorimetric visualization of trinitrotoluene at picomolar levels using gold nanoparticles. , 2008, Angewandte Chemie.

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

[41]  A. Llorente,et al.  Proteomic Analysis of Microvesicles Released by the Human Prostate Cancer Cell Line PC-3 , 2012, Molecular & Cellular Proteomics.

[42]  M. Mason,et al.  Proteomics Analysis of Cancer Exosomes Using a Novel Modified Aptamer-based Array (SOMAscanTM) Platform , 2014, Molecular & Cellular Proteomics.

[43]  Hong Zhao,et al.  Colorimetric detection of glucose in rat brain using gold nanoparticles. , 2010, Angewandte Chemie.

[44]  G. Duigou,et al.  Decreased replication ability of E1-deleted adenoviruses correlates with increased brain tumor malignancy. , 2005, Cancer research.

[45]  E. Klein,et al.  Novel role of prostate-specific membrane antigen in suppressing prostate cancer invasiveness. , 2005, Cancer research.

[46]  Muling Shi,et al.  Aptasensor with Expanded Nucleotide Using DNA Nanotetrahedra for Electrochemical Detection of Cancerous Exosomes. , 2017, ACS nano.

[47]  E. Wang,et al.  Simple and sensitive aptamer-based colorimetric sensing of protein using unmodified gold nanoparticle probes. , 2007, Chemical communications.

[48]  Kevin M. Bradley,et al.  Aptamers against Cells Overexpressing Glypican 3 from Expanded Genetic Systems Combined with Cell Engineering and Laboratory Evolution. , 2016, Angewandte Chemie.

[49]  F. Magni,et al.  Advances in membranous vesicle and exosome proteomics improving biological understanding and biomarker discovery , 2011, Proteomics.

[50]  D. Bigner,et al.  Proteomic and immunologic analyses of brain tumor exosomes , 2009, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[51]  Steven A Carr,et al.  Protein biomarker discovery and validation: the long and uncertain path to clinical utility , 2006, Nature Biotechnology.

[52]  Weihong Tan,et al.  Aptamer-conjugated nanoparticles for cancer cell detection. , 2011, Analytical chemistry.

[53]  C. Chiang,et al.  The General Transcription Machinery and General Cofactors , 2006, Critical reviews in biochemistry and molecular biology.

[54]  Weihong Tan,et al.  Mapping receptor density on live cells by using fluorescence correlation spectroscopy. , 2009, Chemistry.

[55]  Martin J. Hessner,et al.  Transcriptional Signatures as a Disease-Specific and Predictive Inflammatory Biomarker for Type 1 Diabetes , 2012, Genes and Immunity.

[56]  Weian Zhao,et al.  Bioinspired multivalent DNA network for capture and release of cells , 2012, Proceedings of the National Academy of Sciences.

[57]  György Nagy,et al.  Cellular and Molecular Life Sciences REVIEW Membrane vesicles, current state-of-the-art: emerging role of extracellular vesicles , 2022 .

[58]  W. Tan,et al.  Cell-SELEX-based aptamer-conjugated nanomaterials for cancer diagnosis and therapy , 2015 .