Tumor-selective replication herpes simplex virus-based technology significantly improves clinical detection and prognostication of viable circulating tumor cells

// Wen Zhang 1, * , Li Bao 3, 9, 10, * , Shaoxing Yang 6, * , Zhaoyang Qian 3, * , Mei Dong 4, * , Liyuan Yin 2, 19 , Qian Zhao 7 , Keli Ge 1 , Zhenling Deng 1 , Jing Zhang 5 , Fei Qi 8 , Zhongxue An 3 , Yuan Yu 3 , Qingbo Wang 3 , Renhua Wu 3 , Fan Fan 3 , Lianfeng Zhang 16 , Xiping Chen 17 , Yingjian Na 17 , Lin Feng 2 , Lingling liu 1 , Yujie Zhu 1 , Tiancheng Qin 2 , Shuren Zhang 1 , Youhui Zhang 1 , Xiuqing Zhang 3, 11, 12 , Jian Wang 3, 13 , Xin Yi 3 , Liqun Zou 5 , Hong-Wu Xin 18 , Henrik J. Ditzel 10, 14 , Hongjun Gao 6 , Kaitai Zhang 2 , Binlei Liu 15, 1 , Shujun Cheng 2 1 Department of Immunology, Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China 2 State Key Laboratory of Molecular Oncology, Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China 3 BGI-Shenzhen, Shenzhen 518083, China 4 Department of Medical Oncology, Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China 5 Department of Medical Oncology, Cancer Center, West China Hospital of Sichuan University, Chengdu 610041, China 6 Department of Pulmonary Oncology, Affiliated Hospital, Academy of Military Medical Science, Beijing 100071, China 7 Department of Pathology, Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China 8 Department of Respiratory Diseases, Chinese PLA General Hospital, Beijing 100853, China 9 Molecular Disease Biology Section, Department of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, Danish Cancer Society, DK-2100 Copenhagen, Denmark 10 Department of Cancer and Inflammation Research, Institute of Molecular Medicine, University of Southern Denmark, DK-5000 Odense C, Denmark 11 The Guangdong Enterprise Key Laboratory of Human Disease Genomics, BGI-Shenzhen, Shenzhen 518083, China 12 Shenzhen Key Laboratory of Transomics Biotechnologies, BGI-Shenzhen, 518083 Shenzhen, China 13 James D. Watson Institute of Genome Science, 310008 Hangzhou, China 14 Department of Oncology, Odense University Hospital, DK-5000 Odense C, Denmark 15 Hubei Provincial Cooperative, Innovation Center of Industrial Fermentation, Hubei University of Technology, Wuhan 30068, China 16 Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 10021, China 17 Institute of Environmental Health and Related Product Safety, China CDC, Beijing 10021, China 18 Laborartory of Molecular Oncology, Key Laboratory of Carcinogenesis and Translational Research, Ministry of Education, Peking University Cancer Hospital and Institute, Beijing 100142, China 19 Lung Cancer Center, Cancer Center, West China Hospital of Sichuan University, Chengdu 610041, China * These authors contributed equally to this work Correspondence to: Binlei Liu, email: liubinlei@cicams.ac.cn Kaitai Zhang, email: zhangkt@cicams.ac.cn Hongjun Gao, email: gaohj6708@hotmail.com Keywords: viable circulating tumor cells, epithelial-marker-independent, telomerase-specific HSV, clinical application Received: October 20, 2015      Accepted: April 24, 2016      Published: May 18, 2016 ABSTRACT Detection of circulating tumor cells remains a significant challenge due to their vast physical and biological heterogeneity. We developed a cell-surface-marker-independent technology based on telomerase-specific, replication-selective oncolytic herpes-simplex-virus-1 that targets telomerase-reverse-transcriptase-positive cancer cells and expresses green-fluorescent-protein that identifies viable CTCs from a broad spectrum of malignancies. Our method recovered 75.5–87.2% of tumor cells spiked into healthy donor blood, as validated by different methods, including single cell sequencing. CTCs were detected in 59–100% of 326 blood samples from patients with 6 different solid organ carcinomas and lymphomas. Significantly, CTC-positive rates increased remarkably with tumor progression from N0M0, N+M0 to M1 in each of 5 tested cancers (lung, colon, liver, gastric and pancreatic cancer, and glioma). Among 21 non-small cell lung cancer cases in which CTC values were consecutively monitored, 81% showed treatment-related decreases, which was also found after treatments in the other solid tumors. Moreover, monitoring CTC values provided an efficient treatment response indicator in hematological malignancies. Compared to CellSearch, our method detected significantly higher positive rates in 40 NSCLC in all stages, including N0M0, N+M0 and M1, and was less affected by chemotherapy. This simple, robust and clinically-applicable technology detects viable CTCs from solid and hematopoietic malignancies in early to late stages, and significantly improves clinical detection and treatment prognostication.