Volume 147, Issue 2, Pages 426-432 (November 2017) iCTC drug resistance (CDR) Testing ex vivo for evaluation of available therapies to treat patients with epithelial ovarian cancer  Michael L. Pearl, Huan Dong, Qiang Zhao, Shaun Tulley, Marlo K. Dombroff, Wen-Tien Chen  Gynecologic Oncology  Volume 147, Issue 2, Pages 426-432 (November 2017) DOI: 10.1016/j.ygyno.2017.08.018 Copyright © 2017 The Authors Terms and Conditions

Fig. 1 Imaging and counting of iCTCs and immune cells in the CDR assay using blood from an EOC patient. (A). Images were taken after fixed cells were stained with HL markers for immune cells by Differential Interference Contract microscopy. Bars=40μm. (B). Enumeration of CAM-avid iCTCs and immune (HL) cells in 0.5-mL blood using flow cytometry. Gates were made on G-CAM+ (CAM uptake)/7AAD+ fixed cells (G1) to exclude platelets or non-cellular particles (left panels in Control and Gemzar boxes). HL+/dim events were also excluded (events outside of G2). Events that overlap between G1 and G2 are identified and enumerated as iCTC counts shown in red on the right panel of the plot. Note the assay was duplicated. Gemzar (gemcitabine, nucleoside metabolic inhibitor). Gynecologic Oncology 2017 147, 426-432DOI: (10.1016/j.ygyno.2017.08.018) Copyright © 2017 The Authors Terms and Conditions

Fig. 2 Representatives of CDR results using blood from EOC patients before the Taxol-Carbo therapy. (A). CDR scores using 0.5-mL blood from SB448 patient per drug or combination. Each data point represents mean±SD of duplicates. Note: rec., recurrence. (B). CDR scores using 0.5-mL blood from SB483 patient per drug or combination. Each data point represents mean±SD of duplicates. Note: IIIb, stage IIIb. (C). CDR scores using 0.5-mL blood from SB455 patient per drug or combination. Each data point represents mean±SD of duplicates. Note: IIIc, stage IIIc. Generic names of drugs were provided in (): Taxotere (docetaxel, taxane derivatives and anti-microtubule agents), Adriamycin (doxorubicin) and Doxil (doxorubicin liposomal, anthracyclines compounds), Hycamtin (topotecan, topoisomerase I inhibitor), Gemzar (gemcitabine, nucleoside metabolic inhibitor), cyclophosphamide (nitrogen mustard alkylating agent), and combined Taxotere and carboplatin (Taxol-Carbo). Gynecologic Oncology 2017 147, 426-432DOI: (10.1016/j.ygyno.2017.08.018) Copyright © 2017 The Authors Terms and Conditions

Fig. 3 Representatives of treatment monitoring of individual patients using iCTCs and CDR results using blood sampling from patients during and after therapy. (A). Changes in iCTCs (upper plot) and CDR results (lower plot) of blood sampling, marked with red circle, from SB419 patient with stage IV EOC treated with three lines of therapy as indicated in the upper plot. In this case, decrease in iCTCs detected positive response in the first line of therapy; increase in iCTCs identified relapse and non-responsiveness of the second line of chemotherapy and the third line of chemotherapy. CDR scores using 0.5-mL blood per drug or combination. Each data point represents mean±SD of duplicates.). Note: progressive disease (PD); no evidence of disease (NED) using CT scans and/or tumor biopsy. (B). Changes in iCTCs (upper plot) and CDR results (lower plot) of blood sampling, marked with red circle, from SB445 patient with stage IV EOC treated with therapy as indicated in the upper plot. Decrease in iCTCs detected positive response from PD to NED during therapy. CDR scores using 0.5-mL blood per drug or combination. Each data point represents mean±SD of duplicates. Generic names of drugs were provided in (): Taxotere (docetaxel, taxane derivatives and anti-microtubule agents), Adriamycin (doxorubicin) and Doxil (doxorubicin liposomal, anthracyclines compounds), Hycamtin (topotecan, topoisomerase I inhibitor), Gemzar (gemcitabine, nucleoside metabolic inhibitor), cyclophosphamide (nitrogen mustard alkylating agent), and combined Taxotere and carboplatin (Taxol-Carbo). Gynecologic Oncology 2017 147, 426-432DOI: (10.1016/j.ygyno.2017.08.018) Copyright © 2017 The Authors Terms and Conditions