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      Development of DNA Aptamers to Native EpCAM for Isolation of Lung Circulating Tumor Cells from Human Blood

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          Abstract

          We selected DNA aptamers to the epithelial cell adhesion molecule (EpCAM) expressed on primary lung cancer cells isolated from the tumors of patients with non-small cell lung cancer using competitive displacement of aptamers from EpCAM by a corresponding antibody. The resulting aptamers clones showed good nanomolar affinity to EpCAM-positive lung cancer cells. Confocal microscopy imaging and spectral profiling of lung cancer tissues confirmed the same protein target for the aptamers and anti-EpCAM antibodies. Furthermore, the resulted aptamers were successfully applied for isolation and detection of circulating tumor cells in clinical samples of peripheral blood of lung cancer patients.

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          The emerging role of EpCAM in cancer and stem cell signaling.

          Markus Münz, Patrick A. Baeuerle, Olivier Gires (2009)
          Initially discovered as a dominant antigen on colon carcinomas, the epithelial cell adhesion molecule (EpCAM) was considered a mere cell adhesion molecule and reliable surface-binding site for therapeutic antibodies. Recent findings can better explain the relevance of EpCAM's high-level expression on human cancers and cancer propagating cells, and its negative prognostic potential for survival of patients with certain cancers. EpCAM has oncogenic potential and is activated by release of its intracellular domain, which can signal into the cell nucleus by engagement of elements of the wnt pathway.
          Article 0 comments Cited 201 times – based on 0 reviews     Review now
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            Analysis of circulating tumor cells in patients with non-small cell lung cancer using epithelial marker-dependent and -independent approaches.

            Matthew G Krebs, Jian-Mei Hou, Robert Sloane … (2012)
            Epithelial circulating tumor cells (CTCs) are detectable in patients with non-small cell lung cancer (NSCLC). However, epithelial to mesenchymal transition, a widely reported prerequisite for metastasis, may lead to an underestimation of CTC number. We compared directly an epithelial marker-dependent (CellSearch) and a marker-independent (isolation by size of epithelial tumor cells [ISET]) technology platform for the ability to identify CTCs. Molecular characteristics of CTCs were also explored. Paired peripheral blood samples were collected from 40 chemonäive, stages IIIA to IV NSCLC patients. CTCs were enumerated by Epithelial Cell Adhesion Molecule-based immunomagnetic capture (CellSearch, Veridex) and by filtration (ISET, RareCell Diagnostics). CTCs isolated by filtration were assessed by immunohistochemistry for epithelial marker expression (cytokeratins, Epithelial Cell Adhesion Molecule, epidermal growth factor receptor) and for proliferation status (Ki67). CTCs were detected using ISET in 32 of 40 (80%) patients compared with 9 of 40 (23%) patients using CellSearch. A subpopulation of CTCs isolated by ISET did not express epithelial markers. Circulating tumor microemboli (CTM, clusters of ≥ 3 CTCs) were observed in 43% patients using ISET but were undetectable by CellSearch. Up to 62% of single CTCs were positive for the proliferation marker Ki67, whereas cells within CTM were nonproliferative. Both technology platforms detected NSCLC CTCs. ISET detected higher numbers of CTCs including epithelial marker negative tumor cells. ISET also isolated CTM and permitted molecular characterization. Combined with our previous CellSearch data confirming CTC number as an independent prognostic biomarker for NSCLC, we propose that this complementary dual technology approach to CTC analysis allows more complete exploration of CTCs in patients with NSCLC.
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              Frequent EpCam protein expression in human carcinomas.

              Marcel Bundi, Guido Sauter, P Went … (2003)
              Expression of the transmembrane glycoprotein EpCam (epithelial cellular adhesion molecule) occurs in normal epithelium of different organs and was described in carcinomas of various sites. Specific anti-EpCam therapies are now being used in clinical trials. Thus, it is of interest to know which tumor types express or overexpress this protein, and in what frequency. We therefore analyzed EpCam expression by immunohistochemistry on a tissue microarray containing 3900 tissue samples of 134 different histological tumor types and subtypes. EpCam expression was detected in 98 of 131 tumor categories. At least a weak EpCam expression in >10% of tumors was observed in 87 of 131 different tumor categories. Adenocarcinomas of the colon (81%) and pancreas (78%), as well as hormone-refractory adenocarcinomas of the prostate (71%), were identified as particularly promising therapy targets with a high fraction of strongly positive tumors. Most soft-tissue tumors and all lymphomas were EpCam negative. It is concluded that anti-EpCam therapies, if proven to be successful, will have broad applications in a wide variety of carcinomas.
              Article 0 comments Cited 167 times – based on 0 reviews     Review now
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                Author and article information

                Journal
                Journal ID (nlm-ta): Cancers (Basel)
                Journal ID (iso-abbrev): Cancers (Basel)
                Journal ID (publisher-id): cancers
                Title: Cancers
                Publisher: MDPI
                ISSN (Electronic): 2072-6694
                Publication date (Electronic): 12 March 2019
                Publication date Collection: March 2019
                Volume: 11
                Issue: 3
                Electronic Location Identifier: 351
                Affiliations
                [1 ]Federal Research Center “Krasnoyarsk Science Center of the Siberian Branch of the Russian Academy of Science”, Krasnoyarsk 660036, Russia; galina.zamay@ 123456gmail.com (G.S.Z.); olga.kolovskaya@ 123456gmail.com (O.S.K.); cobweb9999@ 123456gmail.com (T.I.I.); tzamay@ 123456yandex.ru (T.N.Z.); d_veprintsev@ 123456mail.ru (D.V.V.); yury.glazyrin@ 123456gmail.com (Y.E.G.)
                [2 ]Laboratory for Biomolecular and Medical Technologies, Krasnoyarsk State Medical University named after prof. V.F. Voino-Yasenecki, Krasnoyarsk 660022, Russia; valllya93@ 123456mail.ru (V.L.G.); garanzha.i.v@ 123456mail.ru (I.I.G.); alexkrat@ 123456mail.ru (A.V.K.)
                [3 ]Krasnoyarsk Regional Clinical Cancer Center named after A.I. Kryzhanovsky, Krasnoyarsk 660133, Russia
                [4 ]Department of Chemistry and Biomolecular Sciences, University of Ottawa, Ottawa, ON K1N 6N5, Canada; ana.gargaun@ 123456gmail.com
                [5 ]Laboratory of Advanced Materials and Technology, Siberian Physical-Technical Institute of Tomsk State University, Tomsk 634050, Russia; 201kiop@ 123456mail.ru (I.N.L.); v_svetlichnyi@ 123456bk.ru (V.A.S.)
                Author notes
                Author information
                https://orcid.org/0000-0002-2826-5751
                https://orcid.org/0000-0003-0514-599X
                Article
                Publisher ID: cancers-11-00351
                DOI: 10.3390/cancers11030351
                PMC ID: 6468627
                PubMed ID: 30871104
                SO-VID: f1cece1a-0854-41c5-b3ce-d322c1cd3f2e
                Copyright © © 2019 by the authors.
                License:

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                Date received : 06 February 2019
                Date accepted : 08 March 2019
                Categories
                Subject: Article

                Keywords: aptamers,selex (systematic evolution of ligands by exponential enrichment),epcam,non-small-cell lung cancer,circulating tumor cells,blood
                Data availability:
                Keywords: aptamers, selex (systematic evolution of ligands by exponential enrichment), epcam, non-small-cell lung cancer, circulating tumor cells, blood

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