title: A liquid biopsy-based method for the detection and quantification of circulating tumor cells in surgical osteosarcoma patients.
abstract: A method for the enumeration and quantification of osteosarcoma (OS) circulating tumor cells (CTCs) is currently not available. A correlation between the number of CTCs and progression-free survival (PFS) has been established for other cancers, but not for OS CTCs. A method was therefore developed for CTC quantification in OS and validated in a prospective cohort of surgical patients with primary and recurrent/metastatic OS (N=23). Human OS cells, acting as CTCs, were enumerated from spiked human peripheral blood (PB) following erythrocyte and leukocyte depletion. The OS cells were quantified microscopically based on aneuploidy and a CK18-/CD45- phenotype. Aneuploidy was assayed by fluorescence in situ hybridization (FISH) using fluorescence-labeled alpha-satellite probes for the centromeres of chromosome (CEP 8). CK18 and CD45 phenotyping was performed with immunocytochemistry. HOS cells in spiked PB could be effectively retrieved with the FISH-based enumeration method, which was subsequently employed in an OS patient cohort. PB of recurrent/metastatic OS patients contained more CTCs than the PB of primary OS patients. OS patients with ≥2 CTCs per 7.5 ml of PB had worse PFS than patients whose PB contained <2 CTCs. In 2 cases, CTCs were present in PB of OS patients with negative X-ray and chest CT scans. In conclusion, our method was able to quantitate CTCs in liquid biopsies of OS patients. The number of CTCs has diagnostic and prognostic value.
authors.affiliation: Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China.
authors.forename: Haoqiang
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China.
authors.forename: Peng
authors.initials: P
authors.lastname: Gao
authors.affiliation: Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China.
authors.forename: Xin
authors.initials: X
authors.lastname: Xiao
authors.affiliation: Department of Experimental Surgery, Academic Medical Center, University of Amsterdam, 1105AZ Amsterdam, The Netherlands.
authors.forename: Michal
authors.initials: M
authors.lastname: Heger
authors.affiliation: Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China.
authors.forename: Lei
authors.initials: L
authors.lastname: Geng
authors.affiliation: Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China.
authors.forename: Bo
authors.initials: B
authors.lastname: Fan
authors.affiliation: Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China.
authors.forename: Yulin
authors.initials: Y
authors.lastname: Yuan
authors.affiliation: Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China.
authors.forename: Chen
authors.initials: C
authors.lastname: Huang
authors.affiliation: Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China.
authors.forename: Guojing
authors.initials: G
authors.lastname: Chen
authors.affiliation: Membrane Biochemistry and Biophysics, Institute of Biomembranes, Utrecht University, 3584CH Utrecht, The Netherlands.
authors.forename: Yao
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Orthopedic Oncology, Tianjin Hospital, Tianjin 300210, P.R. China.
authors.forename: Yongchen
authors.initials: Y
authors.lastname: Hu
authors.affiliation: Department of Orthopedics, The General Hospital of Jinan Military Commanding Region, Jinan, Shandong 250031, P.R. China.
authors.forename: Xiuchun
authors.initials: X
authors.lastname: Yu
authors.affiliation: Department of Orthopedics, The General Hospital of Nanjing Military Commanding Region, Nanjing, Jiangsu 210008, P.R. China.
authors.forename: Sujia
authors.initials: S
authors.lastname: Wu
authors.affiliation: Department of Vascular and Endocrine Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China.
authors.forename: Ling
authors.initials: L
authors.lastname: Wang
authors.affiliation: Department of Orthopedics, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Wang