title: A multiplexed marker-based algorithm for diagnosis of carcinoma of unknown primary using circulating tumor cells.
abstract: Real-time, single-cell multiplex immunophenotyping of circulating tumor cells (CTCs) is hypothesized to inform diagnosis of tissue of origin in patients with carcinoma of unknown primary (CUP). In 20 to 50% of CUP patients, the primary site remains unidentified, presenting a challenge for clinicians in diagnosis and treatment. We developed a post-CellSearch CTC assay using multiplexed Q-dot or DyLight conjugated antibodies with the goal of detecting multiple markers in single cells within a CTC population. We adapted our approach to size-based CTC enrichment protocols for capturing CTCs and subsequent immunofluorescence (IF) using a minimal set of markers to predict the primary sites for common metastatic tumors. The carcinomas are characterized with cytokeratin 7 (CK7), cytokeratin 20 (CK20), thyroid transcription factor 1 (TTF-1), estrogen receptor (ER) or prostate-specific antigen (PSA. IF has been optimized in cultured tumor cells with individual antibodies, then with conjugated antibodies to form a multiplex antibody set. With IF, we evaluated antibodies specific to these 5 markers in lung, breast, colorectal, and prostate cancer cell lines and blood from metastatic prostate and breast cancer patients. This advanced technology provides a noninvasive, diagnostic blood test as an adjunct to routine tissue biopsy. Its further implementation requires prospective clinical testing.
authors.affiliation: Laboratory of Translational Oncology and Experimental Cancer Therapeutics, Division of Hematology-Oncology, Penn State Hershey Cancer Institute, Hershey, PA, USA.|Laboratory of Translational Oncology and Experimental Cancer Therapeutics, Department of Medical Oncology and Molecular Therapeutics Program, Fox Chase Cancer Center, Philadelphia, PA, USA.
authors.forename: Elizabeth M
authors.initials: EM
authors.lastname: Matthew
authors.affiliation: Laboratory of Translational Oncology and Experimental Cancer Therapeutics, Division of Hematology-Oncology, Penn State Hershey Cancer Institute, Hershey, PA, USA.|Laboratory of Translational Oncology and Experimental Cancer Therapeutics, Department of Medical Oncology and Molecular Therapeutics Program, Fox Chase Cancer Center, Philadelphia, PA, USA.
authors.forename: Lanlan
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Department of Pathology, Penn State Milton S. Hershey Medical Center, Hershey, PA, USA.
authors.forename: Zhaohai
authors.initials: Z
authors.lastname: Yang
authors.affiliation: Laboratory of Translational Oncology and Experimental Cancer Therapeutics, Division of Hematology-Oncology, Penn State Hershey Cancer Institute, Hershey, PA, USA.|Laboratory of Translational Oncology and Experimental Cancer Therapeutics, Department of Medical Oncology and Molecular Therapeutics Program, Fox Chase Cancer Center, Philadelphia, PA, USA.
authors.forename: David T
authors.initials: DT
authors.lastname: Dicker
authors.affiliation: Division of Hematology-Oncology, Penn State Milton S. Hershey Medical Center and Penn State Hershey Cancer Institute, Hershey, PA, USA.
authors.forename: Sheldon L
authors.initials: SL
authors.lastname: Holder
authors.affiliation: Laboratory of Translational Oncology and Experimental Cancer Therapeutics, Division of Hematology-Oncology, Penn State Hershey Cancer Institute, Hershey, PA, USA.|Division of Hematology-Oncology, Penn State Milton S. Hershey Medical Center and Penn State Hershey Cancer Institute, Hershey, PA, USA.|The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors.forename: Bora
authors.initials: B
authors.lastname: Lim
authors.affiliation: Department of Biomedical Engineering, Penn State University, University Park, PA, USA.
authors.forename: Ramdane
authors.initials: R
authors.lastname: Harouaka
authors.affiliation: Department of Biomedical Engineering, Penn State University, University Park, PA, USA.
authors.forename: Si-Yang
authors.initials: SY
authors.lastname: Zheng
authors.affiliation: Division of Hematology-Oncology, Penn State Milton S. Hershey Medical Center and Penn State Hershey Cancer Institute, Hershey, PA, USA.
authors.forename: Joseph J
authors.initials: JJ
authors.lastname: Drabick
authors.affiliation: Division of Hematology-Oncology, Penn State Milton S. Hershey Medical Center and Penn State Hershey Cancer Institute, Hershey, PA, USA.
authors.forename: Nicholas E
authors.initials: NE
authors.lastname: Lamparella
authors.affiliation: Division of Hematology-Oncology, Penn State Milton S. Hershey Medical Center and Penn State Hershey Cancer Institute, Hershey, PA, USA.
authors.forename: Cristina I
authors.initials: CI
authors.lastname: Truica
authors.affiliation: Laboratory of Translational Oncology and Experimental Cancer Therapeutics, Division of Hematology-Oncology, Penn State Hershey Cancer Institute, Hershey, PA, USA.|Laboratory of Translational Oncology and Experimental Cancer Therapeutics, Department of Medical Oncology and Molecular Therapeutics Program, Fox Chase Cancer Center, Philadelphia, PA, USA.
authors.forename: Wafik S
authors.initials: WS
authors.lastname: El-Deiry
mesh_terms: D000465:Algorithms; D014408:Biomarkers, Tumor / Q000378:metabolism*; D001943:Breast Neoplasms / Q000175:diagnosis* / Q000378:metabolism; D015179:Colorectal Neoplasms / Q000175:diagnosis* / Q000378:metabolism; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D008175:Lung Neoplasms / Q000175:diagnosis* / Q000378:metabolism; D008297:Male; D009382:Neoplasms, Unknown Primary / Q000145:classification* / Q000175:diagnosis / Q000378:metabolism; D009360:Neoplastic Cells, Circulating / Q000473:pathology*; D011471:Prostatic Neoplasms / Q000175:diagnosis* / Q000378:metabolism
chemical_list: D014408:Biomarkers, Tumor
keywords: carcinoma of unknown primary (CUP); circulating tumor cells (CTCs); diagnosis; immunofluorescence