Home LiteratureArticle Details
PMID: 29624843 Published · ppublish English Evaluation Study Journal Article Research Support, Non-U.S. Gov't

A novel multimarker assay for the phenotypic profiling of circulating tumor cells in hepatocellular carcinoma.

Liver transplantation : official publication of the American Association for the Study of Liver Diseases and the International Liver Transplantation Society ·Vol. 24 ·No. 7 ·2018-00-00 ·Pages 946-960

Court CM, Hou S, Winograd P, Segel NH, Li QW, Zhu Y, Sadeghi S, Finn RS, Ganapathy E, Song M, French SW, Naini BV, Sho S, Kaldas FM, Busuttil RW, Tomlinson JS, Tseng HR, Agopian VG

Abstract

Current clinicopathologic staging systems and serum biomarkers poorly discriminate tumor biology in hepatocellular carcinoma (HCC), with high recurrence rates following curative-intent surgical resection and liver transplantation (LT). Identification of accurate biomarkers for improved prognostication and treatment selection is a critical unmet need. We sought to develop a novel "liquid-biopsy" assay capable of detecting HCC circulating tumor cells (CTCs) and characterizing phenotypic subpopulations with prognostic significance. Using HCC cell lines, a tissue microarray, and human blood samples, an antibody cocktail targeting the cell-surface markers asialoglycoprotein receptor (ASGPR), glypican-3, and epithelial cell adhesion molecule was optimized for HCC CTC capture using the NanoVelcro CTC Assay. The ability of HCC CTCs and vimentin (VIM)-positive CTCs (a subpopulation expressing an epithelial-to-mesenchymal phenotype) to accurately discriminate tumor stage, recurrence, progression, and overall survival (OS) was evaluated in a prospective study of 80 patients. Multimarker capture detected greater numbers of CTCs than any individual antibody alone for both cell line and patient samples (P < 0.001). HCC CTCs were identified in 59/61 (97%) patients, and HCC (median, 6 CTCs) and non-HCC patients (median, 1 CTC; area under the receiver operating characteristic curve [AUROC] = 0.92; P < 0.001; sensitivity = 84.2%; specificity = 88.5%) were accurately discriminated. VIM-positive CTCs accurately discriminated early-stage, LT eligible patients (median, 0 CTCs) from locally advanced/metastatic, LT ineligible patients (median, 6 CTCs; AUROC = 0.89; P = 0.001; sensitivity = 87.1%; specificity = 90.0%), and predicted OS for all patients (hazard ratio [HR], 2.21; P = 0.001), and faster recurrence after curative-intent surgical or locoregional therapy in potentially curable early-stage HCC (HR, 3.14; P = 0.002). In conclusion, we developed a novel multimarker CTC enrichment assay that detects HCC CTCs with high efficiency and accuracy. A phenotypic subpopulation of VIM-positive CTCs appears to signify the presence of aggressive underlying disease and occult metastases and may have important implications for treatment selection. Liver Transplantation 24 946-960 2018 AASLD.

MeSH Terms
Aged Asialoglycoprotein Receptor/analysis,metabolism Biological Assay/methods Biomarkers, Tumor/analysis,metabolism Carcinoma, Hepatocellular/blood,mortality,pathology Cell Line, Tumor Epithelial Cell Adhesion Molecule/analysis,metabolism Female Glypicans/analysis,metabolism Healthy Volunteers Humans Immunoassay/methods Kaplan-Meier Estimate Liquid Biopsy/methods Liver Cirrhosis/blood Liver Neoplasms/blood,mortality,pathology Male Microfluidics/methods Middle Aged Neoplasm Recurrence, Local/diagnosis,mortality Neoplastic Cells, Circulating/metabolism Prognosis Prospective Studies Sensitivity and Specificity Tissue Array Analysis Vimentin/metabolism
Chemicals
Asialoglycoprotein Receptor Biomarkers, Tumor Epithelial Cell Adhesion Molecule GPC3 protein, human Glypicans VIM protein, human Vimentin
Authors & Affiliations
18 authors, click to expand affiliations / ORCID
Court Colin M ORCID
Departments of Surgery, University of California, Los Angeles, Los Angeles, CA. | Department of Molecular, Cellular, and Integrative Physiology, University of California, Los Angeles, Los Angeles, CA. | Department of Surgery, Veteran's Health Administration, Greater Los Angeles, Los Angeles, CA.
Hou Shuang
Departments of Surgery, University of California, Los Angeles, Los Angeles, CA.
Winograd Paul
Departments of Surgery, University of California, Los Angeles, Los Angeles, CA. | Department of Surgery, Veteran's Health Administration, Greater Los Angeles, Los Angeles, CA.
Segel Nicholas H
Departments of Surgery, University of California, Los Angeles, Los Angeles, CA. | Department of Molecular and Medical Pharmacology, University of California, Los Angeles, Los Angeles, CA.
Li Qingyu Wilda
Departments of Surgery, University of California, Los Angeles, Los Angeles, CA. | Department of Molecular and Medical Pharmacology, University of California, Los Angeles, Los Angeles, CA.
Zhu Yazhen
Department of Molecular and Medical Pharmacology, University of California, Los Angeles, Los Angeles, CA. | California NanoSystems Institute, University of California, Los Angeles, Los Angeles, CA.
Sadeghi Saeed
Department of Medicine, Division of Hematology/Oncology, University of California, Los Angeles, Los Angeles, CA.
Finn Richard S
Department of Medicine, Division of Hematology/Oncology, University of California, Los Angeles, Los Angeles, CA.
Ganapathy Ekambaram
Department of Pathology and Laboratory Medicine, University of California, Los Angeles, Los Angeles, CA.
Song Min
Department of Molecular and Medical Pharmacology, University of California, Los Angeles, Los Angeles, CA. | California NanoSystems Institute, University of California, Los Angeles, Los Angeles, CA.
French Samuel W
Department of Pathology and Laboratory Medicine, University of California, Los Angeles, Los Angeles, CA.
Naini Bita V
Department of Pathology and Laboratory Medicine, University of California, Los Angeles, Los Angeles, CA.
Sho Shonan
Departments of Surgery, University of California, Los Angeles, Los Angeles, CA. | Department of Surgery, Veteran's Health Administration, Greater Los Angeles, Los Angeles, CA.
Kaldas Fady M
Departments of Surgery, University of California, Los Angeles, Los Angeles, CA.
Busuttil Ronald W
Departments of Surgery, University of California, Los Angeles, Los Angeles, CA.
Tomlinson James S
Departments of Surgery, University of California, Los Angeles, Los Angeles, CA. | Jonsson Comprehensive Cancer Center, University of California, Los Angeles, Los Angeles, CA. | Department of Surgery, Veteran's Health Administration, Greater Los Angeles, Los Angeles, CA.
Tseng Hsian-Rong
Department of Molecular and Medical Pharmacology, University of California, Los Angeles, Los Angeles, CA. | California NanoSystems Institute, University of California, Los Angeles, Los Angeles, CA. | Jonsson Comprehensive Cancer Center, University of California, Los Angeles, Los Angeles, CA.
Agopian Vatche G
Departments of Surgery, University of California, Los Angeles, Los Angeles, CA. | Jonsson Comprehensive Cancer Center, University of California, Los Angeles, Los Angeles, CA.
References (36)
36 references, click to expand
  1. Digital quantitation of HCC-associated stem cell markers and protein quality control factors using tissue arrays of human liver sections.
    Exp Mol Pathol. 2014 Dec;97(3):399-410 PMID: 25218810
  2. Liver transplantation for hepatocellular carcinoma: expansion of the tumor size limits does not adversely impact survival.
    Hepatology. 2001 Jun;33(6):1394-403 PMID: 11391528
  3. Modified RECIST (mRECIST) assessment for hepatocellular carcinoma.
    Semin Liver Dis. 2010 Feb;30(1):52-60 PMID: 20175033
  4. Association of Vimentin overexpression and hepatocellular carcinoma metastasis.
    Oncogene. 2004 Jan 8;23(1):298-302 PMID: 14647434
  5. Prediction of posthepatectomy recurrence of hepatocellular carcinoma by circulating cancer stem cells: a prospective study.
    Ann Surg. 2011 Oct;254(4):569-76 PMID: 21892074
  6. Individual profiling of circulating tumor cell composition and therapeutic outcome in patients with hepatocellular carcinoma.
    Transl Oncol. 2013 Aug 01;6(4):420-8 PMID: 23908685
  7. Epithelial-mesenchymal transition phenotypes of circulating tumor cells correlate with the clinical stages and cancer metastasis in hepatocellular carcinoma patients.
    Cancer Biomark. 2017 Dec 6;20(4):487-498 PMID: 28869439
  8. Nanostructure embedded microchips for detection, isolation, and characterization of circulating tumor cells.
    Acc Chem Res. 2014 Oct 21;47(10):2941-50 PMID: 25111636
  9. Expression of Ep-CAM in normal, regenerating, metaplastic, and neoplastic liver.
    J Pathol. 1999 Jun;188(2):201-6 PMID: 10398165
  10. Comprehensive analysis of the independent effect of twist and snail in promoting metastasis of hepatocellular carcinoma.
    Hepatology. 2009 Nov;50(5):1464-74 PMID: 19821482
  11. Cutoff Finder: a comprehensive and straightforward Web application enabling rapid biomarker cutoff optimization.
    PLoS One. 2012;7(12):e51862 PMID: 23251644
  12. Circulating tumour cells as a biomarker for diagnosis and staging in pancreatic cancer.
    Br J Cancer. 2016 Jun 14;114(12):1367-75 PMID: 27300108
  13. Global and regional mortality from 235 causes of death for 20 age groups in 1990 and 2010: a systematic analysis for the Global Burden of Disease Study 2010.
    Lancet. 2012 Dec 15;380(9859):2095-128 PMID: 23245604
  14. Imagestream detection and characterisation of circulating tumour cells - A liquid biopsy for hepatocellular carcinoma?
    J Hepatol. 2016 Aug;65(2):305-13 PMID: 27132171
  15. Isolation of circulating tumor cells in patients with hepatocellular carcinoma using a novel cell separation strategy.
    Clin Cancer Res. 2011 Jun 1;17(11):3783-93 PMID: 21527564
  16. Hepatocellular carcinoma: reasons for phase III failure and novel perspectives on trial design.
    Clin Cancer Res. 2014 Apr 15;20(8):2072-9 PMID: 24589894
  17. Circulating epithelial cells in patients with benign colon diseases.
    Clin Chem. 2012 May;58(5):936-40 PMID: 22205690
  18. Detection of circulating tumor cells in hepatocellular carcinoma using antibodies against asialoglycoprotein receptor, carbamoyl phosphate synthetase 1 and pan-cytokeratin.
    PLoS One. 2014 Apr 24;9(4):e96185 PMID: 24763545
  19. Circulating tumor cells from patients with advanced prostate and breast cancer display both epithelial and mesenchymal markers.
    Mol Cancer Res. 2011 Aug;9(8):997-1007 PMID: 21665936
  20. Role of epithelial to mesenchymal transition in hepatocellular carcinoma.
    J Hepatol. 2016 Oct;65(4):798-808 PMID: 27212245
  21. Presence of EpCAM-positive circulating tumor cells as biomarker for systemic disease strongly correlates to survival in patients with hepatocellular carcinoma.
    Int J Cancer. 2013 Nov;133(9):2165-71 PMID: 23616258
  22. Reality of Single Circulating Tumor Cell Sequencing for Molecular Diagnostics in Pancreatic Cancer.
    J Mol Diagn. 2016 Sep;18(5):688-696 PMID: 27375074
  23. Liver transplantation for the treatment of small hepatocellular carcinomas in patients with cirrhosis.
    N Engl J Med. 1996 Mar 14;334(11):693-9 PMID: 8594428
  24. Cell-surface major vault protein promotes cancer progression through harboring mesenchymal and intermediate circulating tumor cells in hepatocellular carcinomas.
    Sci Rep. 2017 Oct 16;7(1):13201 PMID: 29038587
  25. Vimentin in cancer and its potential as a molecular target for cancer therapy.
    Cell Mol Life Sci. 2011 Sep;68(18):3033-46 PMID: 21637948
  26. Comparison of Two Types of Liquid Biopsies in Patients With Hepatocellular Carcinoma Awaiting Orthotopic Liver Transplantation.
    Transplant Proc. 2015 Nov;47(9):2639-42 PMID: 26680058
  27. Glypican-3 expression distinguishes small hepatocellular carcinomas from cirrhosis, dysplastic nodules, and focal nodular hyperplasia-like nodules.
    Am J Surg Pathol. 2006 Nov;30(11):1405-11 PMID: 17063081
  28. Epithelial-mesenchymal transition markers expressed in circulating tumor cells in hepatocellular carcinoma patients with different stages of disease.
    Cell Death Dis. 2013 Oct 03;4:e831 PMID: 24091674
  29. Biofunctionalized magnetic nanospheres-based cell sorting strategy for efficient isolation, detection and subtype analyses of heterogeneous circulating hepatocellular carcinoma cells.
    Biosens Bioelectron. 2016 Nov 15;85:633-640 PMID: 27240010
  30. A novel prognostic nomogram accurately predicts hepatocellular carcinoma recurrence after liver transplantation: analysis of 865 consecutive liver transplant recipients.
    J Am Coll Surg. 2015 Apr;220(4):416-27 PMID: 25690672
  31. Circulating Tumor Cells for Predicting the Prognostic of Patients with Hepatocellular Carcinoma: A Meta Analysis.
    Cell Physiol Biochem. 2015;37(2):629-40 PMID: 26344495
  32. Reporting recommendations for tumor marker prognostic studies (REMARK).
    J Natl Cancer Inst. 2005 Aug 17;97(16):1180-4 PMID: 16106022
  33. Circulating breast tumor cells exhibit dynamic changes in epithelial and mesenchymal composition.
    Science. 2013 Feb 1;339(6119):580-4 PMID: 23372014
  34. Identification of a population of blood circulating tumor cells from breast cancer patients that initiates metastasis in a xenograft assay.
    Nat Biotechnol. 2013 Jun;31(6):539-44 PMID: 23609047
  35. Correlation Between Postoperative Early Recurrence of Hepatocellular Carcinoma and Mesenchymal Circulating Tumor Cells in Peripheral Blood.
    J Gastrointest Surg. 2018 Apr;22(4):633-639 PMID: 29159757
  36. Subclassification of prostate cancer circulating tumor cells by nuclear size reveals very small nuclear circulating tumor cells in patients with visceral metastases.
    Cancer. 2015 Sep 15;121(18):3240-51 PMID: 25975562
Article Info
Journal
Liver transplantation : official publication of the American Association for the Study of Liver Diseases and the International Liver Transplantation Society
Abbr.
Liver Transpl
ISSN
1527-6473
DOI
10.1002/lt.25062
Published
2018-00-00
Pages
946-960
Language
English
Region
United States
NLM ID
100909185
PMCID
PMC6097911
Subset
IM
Grants
NCI NIH HHS · R21 CA216807 · United States
NCI NIH HHS · R33 CA174562 · United States
NCI NIH HHS · P30 CA016042 · United States
NCI NIH HHS · R44 CA180482 · United States
NCI NIH HHS · U01 CA198900 · United States
NCI NIH HHS · R01 CA246304 · United States
External Links
PubMed source DOI: 10.1002/lt.25062 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2