Electrophysiology-based stratification of pancreatic tumorigenicity by label-free single-cell impedance cytometry
Pancreatic ductal adenocarcinoma (PDAC) is an aggressive cancer lacking specific biomarkers that can be correlated to disease onset, promotion and progression. To assess whether tumor cell electrophysiology may serve as a marker for PDAC tumorigenicity, we use multi-frequency impedance cytometry at...
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| Published in: | Analytica chimica acta Vol. 1101; pp. 90 - 98 |
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| Main Authors: | , , , , , , , , , , , |
| Format: | Journal Article |
| Language: | English |
| Published: |
Netherlands
Elsevier B.V
08-03-2020
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| Subjects: | |
| Online Access: | Get full text |
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| Summary: | Pancreatic ductal adenocarcinoma (PDAC) is an aggressive cancer lacking specific biomarkers that can be correlated to disease onset, promotion and progression. To assess whether tumor cell electrophysiology may serve as a marker for PDAC tumorigenicity, we use multi-frequency impedance cytometry at high throughput (∼350 cells/s) to measure the electrical phenotype of single PDAC tumor cells from xenografts, which are derived from primary pancreatic tumors versus those from liver metastases of different patients. A novel phase contrast metric based on variations in the high and low frequency impedance phase responses that is related to electrophysiology of the cell interior is found to be systematically altered as a function of tumorigenicity. PDAC cells of higher tumorigenicity exhibited lowered interior conductivity and enhanced permittivity, which is validated by the dielectrophoresis on the respective cell types. Using genetic analysis, we suggest the role of dysregulated Na+ transport and removal of Ca2+ ions from the cytoplasm on key oncogenic KRAS-driven processes that may be responsible for lowering of the interior cell conductivity. We envision that impedance cytometry can serve as a tool to quantify phenotypic heterogeneity for rapidly stratifying tumorigenicity. It can also aid in protocols for dielectrophoretic isolation of cells with a particular phenotype for prognostic studies on patient survival and to tailor therapy selection to specific patients.
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•High-throughput (∼350 cells/s) impedance cytometry to measure ∼10,000 cells/sample.•Samples of patient-derived pancreatic tumor xenografts of varying tumorigenicity.•Impedance phase due to electrophysiology of cell interior varies with tumorigenicity.•Lowered interior conductivity and enhanced permittivity for cells of higher tumorigenicity.•Validated based on level of positive dielectrophoresis and its crossover frequency. |
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| Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Authors contributed equally JSM conceived the method, designed, fabricated and integrated the system, designed/ performed the experiments, participated in analyzing the data and generating the manuscript. CH designed/ performed the experiments, analyzed/ interpreted the data, produced the figures and participated in generating the manuscript. JHM performed sample preparation, designed/ performed experiments and participated in data analysis/ interpretation. SJA, EMB, BGB and SN participated in the sample choice for the patient-derived cell lines. WBV, AS and VF participated in fabricating the system. SJA and EMB participated in analyzing/ interpreting the data. SN and EMB participated in revising the manuscript. TWB developed all the sample sets and NSS conceived application of the method to the samples, oversaw the team on experimental design and the manuscript revisions. Electronic Supplementary Information available Author Contributions |
| ISSN: | 0003-2670 1873-4324 |
| DOI: | 10.1016/j.aca.2019.12.033 |