Xiaosai Yao1, Christina Williamson2, Viktor A Adalsteinsson3, Richard S D'Agostino2, Torin Fitton2, Gregory G Smaroff2, Robert T William4, K Dane Wittrup5, J Christopher Love6. 1. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Mass; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Mass. 2. Department of Thoracic and Cardiovascular Surgery, Lahey Hospital and Medical Center, Burlington, Mass. 3. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Mass; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Mass; Broad Institute of MIT and Harvard, Cambridge, Mass. 4. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Mass. 5. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Mass; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Mass; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Mass. 6. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Mass; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Mass; Broad Institute of MIT and Harvard, Cambridge, Mass. Electronic address: clove@mit.edu.
Abstract
OBJECTIVE: Intraoperative tumor shedding may facilitate tumor dissemination. In earlier studies, shed tumor cells were defined primarily by cytomorphological examination, and normal epithelial cells could not always be distinguished from tumor cells. We sought to accurately identify tumor cells using single-cell sequencing and determine whether these cells were mobilized into the circulation during pulmonary lobectomy. METHODS: Forty-two blood samples collected from the tumor-draining pulmonary vein at the end of lobectomy procedures were analyzed. Arrays of nanowells were used to enumerate and retrieve single EpCAM(+) cells. Targeted sequencing of 10 to 15 cells and nested polymerase chain reaction of single cells detected somatic mutations in shed epithelial cells consistent with patient-matched tumor but not normal tissue. RESULTS: The mean number of EpCAM(+) cells in video-assisted thoracoscopy (VATS) lobectomy (no wedge) specimens (n = 16) was 165 (median, 115; range, 0-509) but sampling cells from 3 patients indicated that only 0% to 38% of the EpCAM(+) cells were tumor cells. The mean number of EpCAM(+) cells in VATS lobectomy (wedge) specimens (n = 12) was 1128 (median, 197; range, 47-9406) and all of the EpCAM(+) cells were normal epithelial cells in 2 patients sampled. The mean number of EpCAM(+) cells in thoracotomy specimens (n = 14) was 238 (median, 22; range, 9-2920) and 0% to 50% of total EpCAM(+) cells were tumor cells based on 4 patients sampled. CONCLUSIONS: Surgery mobilizes tumor cells into the pulmonary vein, along with many normal epithelial cells. EpCAM alone cannot differentiate between normal and tumor cells. On the other hand, single-cell genetic approaches with patient-matched normal and tumor tissues can accurately quantify the number of shed tumor cells.
OBJECTIVE:Intraoperative tumor shedding may facilitate tumor dissemination. In earlier studies, shed tumor cells were defined primarily by cytomorphological examination, and normal epithelial cells could not always be distinguished from tumor cells. We sought to accurately identify tumor cells using single-cell sequencing and determine whether these cells were mobilized into the circulation during pulmonary lobectomy. METHODS: Forty-two blood samples collected from the tumor-draining pulmonary vein at the end of lobectomy procedures were analyzed. Arrays of nanowells were used to enumerate and retrieve single EpCAM(+) cells. Targeted sequencing of 10 to 15 cells and nested polymerase chain reaction of single cells detected somatic mutations in shed epithelial cells consistent with patient-matched tumor but not normal tissue. RESULTS: The mean number of EpCAM(+) cells in video-assisted thoracoscopy (VATS) lobectomy (no wedge) specimens (n = 16) was 165 (median, 115; range, 0-509) but sampling cells from 3 patients indicated that only 0% to 38% of the EpCAM(+) cells were tumor cells. The mean number of EpCAM(+) cells in VATS lobectomy (wedge) specimens (n = 12) was 1128 (median, 197; range, 47-9406) and all of the EpCAM(+) cells were normal epithelial cells in 2 patients sampled. The mean number of EpCAM(+) cells in thoracotomy specimens (n = 14) was 238 (median, 22; range, 9-2920) and 0% to 50% of total EpCAM(+) cells were tumor cells based on 4 patients sampled. CONCLUSIONS: Surgery mobilizes tumor cells into the pulmonary vein, along with many normal epithelial cells. EpCAM alone cannot differentiate between normal and tumor cells. On the other hand, single-cell genetic approaches with patient-matched normal and tumor tissues can accurately quantify the number of shed tumor cells.
Authors: J Christopher Love; Jehnna L Ronan; Gijsbert M Grotenbreg; Annemarthe G van der Veen; Hidde L Ploegh Journal: Nat Biotechnol Date: 2006-05-14 Impact factor: 54.908
Authors: Moritz Koch; Peter Kienle; Ulf Hinz; Dalibor Antolovic; Jan Schmidt; Christian Herfarth; Magnus von Knebel Doeberitz; Jürgen Weitz Journal: Ann Surg Date: 2005-02 Impact factor: 12.969
Authors: Daniel Förnvik; Ingvar Andersson; Magnus Dustler; Roy Ehrnström; Lisa Rydén; Anders Tingberg; Sophia Zackrisson; Kristina Aaltonen Journal: Breast Cancer Res Treat Date: 2013-08-29 Impact factor: 4.872
Authors: Jens G Lohr; Viktor A Adalsteinsson; Kristian Cibulskis; Atish D Choudhury; Mara Rosenberg; Peter Cruz-Gordillo; Joshua M Francis; Cheng-Zhong Zhang; Alex K Shalek; Rahul Satija; John J Trombetta; Diana Lu; Naren Tallapragada; Narmin Tahirova; Sora Kim; Brendan Blumenstiel; Carrie Sougnez; Alarice Lowe; Bang Wong; Daniel Auclair; Eliezer M Van Allen; Mari Nakabayashi; Rosina T Lis; Gwo-Shu M Lee; Tiantian Li; Matthew S Chabot; Amy Ly; Mary-Ellen Taplin; Thomas E Clancy; Massimo Loda; Aviv Regev; Matthew Meyerson; William C Hahn; Philip W Kantoff; Todd R Golub; Gad Getz; Jesse S Boehm; J Christopher Love Journal: Nat Biotechnol Date: 2014-04-20 Impact factor: 54.908
Authors: Stefanie S Jeffrey; Elodie Sollier-Christen; Haiyan E Liu; Melanie Triboulet; Amin Zia; Meghah Vuppalapaty; Evelyn Kidess-Sigal; John Coller; Vanita S Natu; Vida Shokoohi; James Che; Corinne Renier; Natalie H Chan; Violet R Hanft Journal: NPJ Genom Med Date: 2017-11-01 Impact factor: 8.617