Dielectrophoretic microfluidic chip enables single-cell measurements for multidrug resistance in heterogeneous acute myeloid leukemia patient samples

Khamenehfar, Avid, Gandhi, Maher K., Chen, Yuchun, Hogge, Donna E. and Li, Paul C. H. (2016) Dielectrophoretic microfluidic chip enables single-cell measurements for multidrug resistance in heterogeneous acute myeloid leukemia patient samples. Analytical Chemistry, 88 11: 5680-5688. doi:10.1021/acs.analchem.5b04446


Author Khamenehfar, Avid
Gandhi, Maher K.
Chen, Yuchun
Hogge, Donna E.
Li, Paul C. H.
Title Dielectrophoretic microfluidic chip enables single-cell measurements for multidrug resistance in heterogeneous acute myeloid leukemia patient samples
Journal name Analytical Chemistry   Check publisher's open access policy
ISSN 1520-6882
0003-2700
Publication date 2016-05-05
Year available 2016
Sub-type Article (original research)
DOI 10.1021/acs.analchem.5b04446
Open Access Status Not Open Access
Volume 88
Issue 11
Start page 5680
End page 5688
Total pages 9
Place of publication Washington, DC, United States
Publisher American Chemical Society
Language eng
Formatted abstract
The front-line treatment for adult acute myeloid leukemia (AML) is anthracycline-based combination chemotherapy. However, treatment outcomes remain suboptimal with relapses frequently observed. Among the mechanisms of treatment failure is multidrug resistance (MDR) mediated by the ABCB1, ABCC1, and ABCG2 drug-efflux transporters. Although genetic and phenotypic heterogeneity between leukemic blast cells is a well-recognized phenomenon, there remains minimal data on differences in MDR activity at the individual cell level. Specifically, functional assays that can distinguish the variability in MDR activity between individual leukemic blasts are lacking. Here, we outline a new dielectrophoretic (DEP) chip-based assay. This assay permits measurement of drug accumulation in single cells, termed same-single-cell analysis in the accumulation mode (SASCA-A). Initially, the assay was optimized in pretherapy samples from 20 adults with AML whose leukemic blasts had MDR activity against the anthracyline daunorubicin (DNR) tested using multiple MDR inhibitors. Parameters tested were initial drug accumulation, time to achieve signal saturation, fold-increase of DNR accumulation with MDR inhibition, ease of cell trapping, and ease of maintaining the trapped cells stationary. This enabled categorization into leukemic blast cells with MDR activity (MDR+) and leukemic blast cells without MDR activity (MDR-ve). Leukemic blasts could also be distinguished from benign white blood cells (notably these also lacked MDR activity). MDR-ve blasts were observed to be enriched in samples taken from patients who went on to enter complete remission (CR), whereas MDR+ blasts were frequently observed in patients who failed to achieve CR following front-line chemotherapy. However, pronounced variability in functional MDR activity between leukemic blasts was observed, with MDR+ cells not infrequently seen in some patients that went on to achieve CR. Next, we tested MDR activity in two paired AML patient samples. Pretherapy samples taken from patients that achieved CR to front-line chemotherapy were compared with samples taken at time of subsequent relapse. MDR+ cells were frequently observed in leukemic blast cells in both pretherapy and relapsed samples, consistent with MDR as a mechanism of relapse in these patients. We demonstrate the ability of a new DEP microfluidic chip-based assay to identify heterogeneity in MDR activity in leukemic blasts. The test provides a platform for future studies to characterize the mechanistic basis for heterogeneity in MDR activity at the individual cell level.
Keyword Adult acute myeloid leukemia
AML
Multidrug resistance
MDR
Dielectrophoretic chip-based assay
Leukemic blast cells
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: HERDC Pre-Audit
UQ Diamantina Institute Publications
 
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