Enhanced detection and comprehensive in situ phenotypic characterization of circulating and disseminated heteroploid epithelial and glioma tumor cells.
Conventional strategy of anti-EpCAM capture and immunostaining of cytokeratins (CKs) to detect circulating tumor cells (CTCs) is limited by highly heterogeneous and dynamic expression or absence of EpCAM and/or CKs in CTCs. In this study, a novel integrated cellular and molecular approach of subtraction enrichment (SE) and immunostaining-FISH (iFISH) was successfully developed. Both large or small size CTCs and circulating tumor microemboli (CTM) in various biofluid samples including cerebrospinal fluid (CSF) of cancer patients and patient-derived-xenograft (PDX) mouse models were efficiently enriched and comprehensively identified and characterized by SE-iFISH. Non-hematopoietic CTCs with heteroploid chromosome 8 were detected in 87-92% of lung, esophageal and gastric cancer patients. Characterization of CTCs performed by CK18-iFISH showed that CK18, the dual epithelial marker and tumor biomarker, was strong positive in only 14% of lung and 24% of esophageal CTCs, respectively. Unlike conventional methodologies restricted only to the large and/or both EpCAM and CK positive CTCs, SE-iFISH enables efficient enrichment and performing in situ phenotypic and karyotypic identification and characterization of the highly heterogeneous CTC subtypes classified by both chromosome ploidy and the expression of various tumor biomarkers. Each CTC subtype may possess distinct clinical significance relative to tumor metastasis, relapse, therapeutic drug sensitivity or resistance, etc.
Department of Thoracic Surgery, Capital Medical University School of Oncology and Chaoyang Hospital, Beijing, China.
Feng
F
Ge
Department of Neurosurgery, Huashan Hospital, Fudan University, Shanghai, China.
Haishi
H
Zhang
Cytelligen, San Diego, California, USA.
Daisy Dandan
DD
Wang
Cytelligen, San Diego, California, USA.
Linda
L
Li
Cytelligen, San Diego, California, USA.
Peter Ping
PP
Lin
D000818:Animals; D000951:Antigens, Neoplasm / Q000378:metabolism; D014408:Biomarkers, Tumor / Q000378:metabolism; D001932:Brain Neoplasms / Q000378:metabolism*; D015815:Cell Adhesion Molecules / Q000378:metabolism; D019008:Drug Resistance, Neoplasm; D000071858:Epithelial Cell Adhesion Molecule; D004847:Epithelial Cells / Q000166:cytology*; D020869:Gene Expression Profiling; D005910:Glioma / Q000378:metabolism*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007621:Karyotyping; D053538:Keratin-18 / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D009364:Neoplasm Recurrence, Local / Q000235:genetics; D009368:Neoplasm Transplantation; D009369:Neoplasms / Q000134:cerebrospinal fluid; D009360:Neoplastic Cells, Circulating / Q000473:pathology; D010641:Phenotype; D010996:Pleural Effusion; D011003:Ploidies
D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D015815:Cell Adhesion Molecules; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule; D053538:Keratin-18
CTC and DTC subtypes; cytokeratin (CK) 18; iFISH; in situ phenotyping and karyotyping; subtraction enrichment