Acquisition of Portal Venous Circulating Tumor Cells From Patients With Pancreaticobiliary Cancers by Endoscopic Ultrasound.
OBJECTIVE
Tumor cells circulate in low numbers in peripheral blood; their detection is used predominantly in metastatic disease. We evaluated the feasibility and safety of sampling portal venous blood via endoscopic ultrasound (EUS) to count portal venous circulating tumor cells (CTCs), compared with paired peripheral CTCs, in patients with pancreaticobiliary cancers (PBCs).


METHODS
In a single-center cohort study, we evaluated 18 patients with suspected PBCs. Under EUS guidance, a 19-gauge EUS fine needle was advanced transhepatically into the portal vein and as many as four 7.5-mL aliquots of blood were aspirated. Paired peripheral blood samples were obtained. Epithelial-derived CTCs were sorted magnetically based on expression of epithelial cell adhesion molecules; only those with a proper morphology and found to be CD45 negative and positive for cytokeratins 8, 18, and/or 19 and 4',6-diamidino-2-phenylindole were considered to be CTCs. For 5 samples, CTCs also were isolated by flow cytometry and based on CD45 depletion. ImageStream was used to determine the relative protein levels of P16, SMAD4, and P53. DNA was extracted from CTCs for sequencing of select KRAS codons.


RESULTS
There were no complications from portal vein blood acquisition. We detected CTCs in portal vein samples from all 18 patients (100%) vs peripheral blood samples from only 4 patients (22.2%). Patients with confirmed PBCs had a mean of 118.4 ± 36.8 CTCs/7.5 mL portal vein blood, compared with a mean of 0.8 ± 0.4 CTCs/7.5 mL peripheral blood (P < .01). The 9 patients with nonmetastatic, resectable, or borderline-resectable PBCs had a mean of 83.2 CTCs/7.5 mL portal vein blood (median, 62.0 CTCs/7.5 mL portal vein blood). In a selected patient, portal vein CTCs were found to carry the same mutations as those detected in a metastatic lymph node and expressed similar levels of P16, SMAD4, and P53 proteins.


CONCLUSIONS
It is feasible and safe to collect portal venous blood from patients undergoing EUS. We identified CTCs in all portal vein blood samples from patients with PBCs, but less than 25% of peripheral blood samples. Portal vein CTCs can be used for molecular characterization of PBCs and share features of metastatic tissue. This technique might be used to study the pathogenesis and progression of PBCs, as well as a diagnostic or prognostic tool to stratify risk of cancer recurrence or developing metastases.
Department of Medicine, Section of Hematology/Oncology, The University of Chicago Medicine and Biological Sciences, Chicago, Illinois.
Daniel V T
DV
Catenacci
Center for Endoscopic Research and Therapeutics, The University of Chicago Medicine and Biological Sciences, Chicago, Illinois.
Christopher G
CG
Chapman
Department of Medicine, Section of Hematology/Oncology, The University of Chicago Medicine and Biological Sciences, Chicago, Illinois.
Peng
P
Xu
Center for Endoscopic Research and Therapeutics, The University of Chicago Medicine and Biological Sciences, Chicago, Illinois.
Ann
A
Koons
Center for Endoscopic Research and Therapeutics, The University of Chicago Medicine and Biological Sciences, Chicago, Illinois.
Vani J
VJ
Konda
Center for Endoscopic Research and Therapeutics, The University of Chicago Medicine and Biological Sciences, Chicago, Illinois.
Uzma D
UD
Siddiqui
Center for Endoscopic Research and Therapeutics, The University of Chicago Medicine and Biological Sciences, Chicago, Illinois. Electronic address: iwaxman@uchicago.edu.
Irving
I
Waxman
D000368:Aged; D000369:Aged, 80 and over; D001661:Biliary Tract Neoplasms / Q000097:blood / Q000235:genetics / Q000473:pathology*; D014408:Biomarkers, Tumor / Q000097:blood / Q000235:genetics; D002452:Cell Count; D002641:Chicago; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000097:blood; D004252:DNA Mutational Analysis; D061765:Endoscopic Ultrasound-Guided Fine Needle Aspiration; D005240:Feasibility Studies; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D018189:Immunomagnetic Separation; D007633:Keratins / Q000097:blood; D017493:Leukocyte Common Antigens / Q000097:blood; D008297:Male; D008875:Middle Aged; D009154:Mutation; D009360:Neoplastic Cells, Circulating / Q000737:chemistry / Q000473:pathology*; D010190:Pancreatic Neoplasms / Q000097:blood / Q000235:genetics / Q000473:pathology*; D011169:Portal Vein / Q000473:pathology*; D011237:Predictive Value of Tests; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics; D051901:Smad4 Protein / Q000097:blood; D016159:Tumor Suppressor Protein p53 / Q000097:blood
D014408:Biomarkers, Tumor; D019941:Cyclin-Dependent Kinase Inhibitor p16; C117307:KRAS protein, human; C497796:SMAD4 protein, human; D051901:Smad4 Protein; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D007633:Keratins; D017493:Leukocyte Common Antigens; C516769:PTPRC protein, human; D016283:Proto-Oncogene Proteins p21(ras)
Endoscopic Ultrasound; EpCAM; Pancreatic Cancer; Portal Venous Blood