Ultra-sensitive liquid biopsy of circulating extracellular vesicles using ExoScreen

Cancer cells secrete small membranous extracellular vesicles (EVs) into their microenvironment and circulation. Although their potential as cancer biomarkers has been promising, the identification and quantification of EVs in clinical samples remains challenging. Here we describe a sensitive and rapid analytical technique for profiling circulating EVs directly from blood samples of patients with colorectal cancer. EVs are captured by two types of antibodies and are detected by photosensitizer-beads, which enables us to detect cancer-derived EVs without a purification step. We also show that circulating EVs can be used for detection of colorectal cancer using the antigen CD147, which is embedded in cancer-linked EVs. This work describes a new liquid biopsy technique to sensitively detect disease-specific circulating EVs and provides perspectives in translational medicine from the standpoint of diagnosis and therapy.

[1]  Hannes Stockinger,et al.  Cancer-related issues of CD147. , 2010, Cancer genomics & proteomics.

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

[3]  Y. Gho,et al.  Quantitative proteomics of extracellular vesicles derived from human primary and metastatic colorectal cancer cells , 2012, Journal of extracellular vesicles.

[4]  Gema Moreno-Bueno,et al.  Melanoma exosomes educate bone marrow progenitor cells toward a pro-metastatic phenotype through MET , 2012, Nature Medicine.

[5]  N. Kosaka,et al.  Comparative marker analysis of extracellular vesicles in different human cancer types , 2013, Journal of extracellular vesicles.

[6]  N. Armitage,et al.  Preoperative carcinoembryonic antigen is related to tumour stage and long-term survival in colorectal cancer. , 1998, British Journal of Cancer.

[7]  Aled Clayton,et al.  Cancer exosomes trigger fibroblast to myofibroblast differentiation. , 2010, Cancer research.

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

[9]  R. Eglen,et al.  The Use of AlphaScreen Technology in HTS: Current Status , 2008, Current chemical genomics.

[10]  Takahiro Ochiya,et al.  Circulating microRNA in body fluid: a new potential biomarker for cancer diagnosis and prognosis , 2010, Cancer science.

[11]  T. Muramatsu,et al.  Basigin (CD147): a multifunctional transmembrane protein involved in reproduction, neural function, inflammation and tumor invasion. , 2003, Histology and histopathology.

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

[13]  G. Parmiani,et al.  Tumour-released exosomes and their implications in cancer immunity , 2008, Cell Death and Differentiation.

[14]  F. Reyal,et al.  Rab27a supports exosome-dependent and -independent mechanisms that modify the tumor microenvironment and can promote tumor progression. , 2012, Cancer research.

[15]  H. Wieand,et al.  Accuracy of fecal occult blood screening for colorectal neoplasia. A prospective study using Hemoccult and HemoQuant tests. , 1993, JAMA.

[16]  J. Nesland,et al.  EMMPRIN is associated with S100A4 and predicts patient outcome in colorectal cancer , 2012, British Journal of Cancer.

[17]  Jing Xu,et al.  HAb18G (CD147), a cancer‐associated biomarker and its role in cancer detection , 2009, Histopathology.

[18]  S. Mathivanan,et al.  ExoCarta: A compendium of exosomal proteins and RNA , 2009, Proteomics.

[19]  G. Sauter,et al.  High incidence of EMMPRIN expression in human tumors , 2006, International journal of cancer.

[20]  A. Guha,et al.  Intercellular transfer of the oncogenic receptor EGFRvIII by microvesicles derived from tumour cells , 2008, Nature Cell Biology.

[21]  Cicek Gercel-Taylor,et al.  MicroRNA signatures of tumor-derived exosomes as diagnostic biomarkers of ovarian cancer. , 2008, Gynecologic oncology.

[22]  Yusuke Yoshioka,et al.  Neutral Sphingomyelinase 2 (nSMase2)-dependent Exosomal Transfer of Angiogenic MicroRNAs Regulate Cancer Cell Metastasis* , 2013, The Journal of Biological Chemistry.

[23]  O. Mäkelä,et al.  Comparison of CA 19-9 and carcinoembryonic antigen (CEA) levels in the serum of patients with colorectal diseases. , 1984, British Journal of Cancer.

[24]  S. Rai,et al.  Identifying mRNA, MicroRNA and Protein Profiles of Melanoma Exosomes , 2012, PloS one.