Clinical significance of detecting CSF-derived tumor cells in breast cancer patients with leptomeningeal metastasis

Abstract

// Xuelu Li 1, 2, * , Yuan Zhang 1, 3, * , Jinlei Ding 1, 4, * , Min Wang 1, 3 , Na Li 1, 5 , Hui Yang 1, 5 , Kainan Wang 1, 5 , Dandan Wang 6 , Peter Ping Lin 6 , Man Li 1, 5 , Zuowei Zhao 1, 2 and Pixu Liu 1, 3, 7 1 The Second Affiliated Hospital, Institute of Cancer Stem Cell, Dalian Medical University, Dalian, China 2 Department of Breast Surgery, The Second Affiliated Hospital, Dalian Medical University, Dalian, China 3 Institute of Cancer Stem Cell, Cancer Center, Dalian Medical University, Dalian, China 4 Cancer Institute, The Second Affiliated Hospital, Dalian Medical University, Dalian, China 5 Department of Oncology, The Second Affiliated Hospital, Dalian Medical University, Dalian, China 6 Cytelligen, San Diego, California, USA 7 College of Pharmacy, Dalian Medical University, Dalian, China * These authors contributed equally to this work Correspondence to: Pixu Liu, email: pixu_liu@dmu.edu.cn Zuowei Zhao, email: dmuzhaozuowei@163.com Man Li, email: dmuliman@163.com Peter Ping Lin, email: plin@cytelligen.com Keywords: breast cancer; leptomeningeal metastasis; cerebrospinal fluid-derived tumor cell; SE-i•FISH Received: July 20, 2017      Accepted: December 14, 2017      Published: December 21, 2017 ABSTRACT Despite marked advances in breast cancer therapy, breast cancer-associated leptomeningeal metastasis (LM), a particularly aggressive syndrome with multifocal seeding of the leptomeninges by tumor cells, still carries an abysmal prognosis. A major problem with breast cancer LM surveillance is the lack of an effective and sensitive means to track dynamic changes of the disease. Cytology detection of cerebrospinal fluid (CSF) is considered the gold standard for LM diagnosis but has a high false-negative rate with a limited sensitivity. Here we applied subtraction enrichment and immunostaining-fluorescence in situ (SE-i•FISH) method, a technique previously used for isolating circulating tumor cells (CTCs) from the peripheral blood, to detect, enumerate, and track cerebrospinal fluid-derived tumor cells (CSFTCs) in CSF samples from 8 breast cancer patients. Comparing with cytology test, we found SE-i•FISH method can accurately and feasibly detect CSFTCs for the diagnosis of breast cancer-associated LM and monitor the disease progression. We also isolated and cultured CSFTCs from these cancer patients and performed genomic sequencing on CSFTCs of two patients. Genomic analysis of CSFTCs against corresponding archival primary breast tumors revealed clonal relationships with some ongoing evolution. Further drug sensitivity test on cultured CSFTCs based on genomic analysis data helped identify promising treatment options for the patient tested. Together, our results suggest that CSFTCs detection using SE-i•FISH platform could serve as a sensitive and accurate method to make the diagnosis and a promising approach to monitor tumor dynamics and treatment response for breast cancer-associated LM.