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Same-single-cell analysis using the microfluidic biochip to reveal drug accumulation enhancement by an amphiphilic diblock copolymer drug formulation

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Abstract

Multidrug resistance (MDR) is one of the major obstacles in drug delivery, and it is usually responsible for unsuccessful cancer treatment. MDR may be overcome by using MDR inhibitors. Among different classes of these inhibitors that block drug efflux mediated by permeability-glycoprotein (P-gp), less toxic amphiphilic diblock copolymers composed of methoxypolyethyleneglycol-block-polycaprolactone (MePEG-b-PCL) have been studied extensively. The purpose of this work is to evaluate how these copolymer molecules can reduce the efflux, thereby enhancing the accumulation of P-gp substrates (e.g., daunorubicin or DNR) in MDR cells. Using conventional methods, it was found that the low-molecular-weight diblock copolymer, MePEG17-b-PCL5 (PCL5), enhanced drug accumulation in MDCKII-MDR1 cells, but the high-molecular-weight version, MePEG114-b-PCL200 (PCL200), did not. However, when PCL200 was mixed with PCL5 (and DNR) in order to encapsulate them to facilitate drug delivery, there was no drug enhancement effect attributable to PCL5, and the reason for this negative result was unclear. Since drug accumulation measured on different cell batches originated from single cells, we employed the same-single-cell analysis in the accumulation mode (SASCA-A) to find out the reason. A microfluidic biochip was used to select single MDR cells, and the accumulation of DNR was fluorescently measured in real time on these cells in the absence and presence of PCL5. The SASCA-A method allowed us to obtain drug accumulation information faster in comparison to conventional assays. The SASCA-A results, and subsequent curve-fitting analysis of the data, have confirmed that when PCL5 was encapsulated in PCL200 nanoparticles as soon as they were synthesized, the ability of PCL5 to enhance DNR accumulation was retained, thus suggesting PCL200 as a promising delivery system for encapsulating P-gp inhibitors, such as PCL5.

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Abbreviations

CsA:

Cyclosporine A

DMEM:

Dulbecco’s modified Eagle's medium

DMSO:

Dimethyl sulfoxide

DNR:

Daunorubicin

DOX:

Doxurubicin

FBS:

Fetal bovine serum

HBSS:

Hanks’ balanced salt solution

MDCK:

Madin-Darby canine kidney

MDR:

Multidrug resistance

MePEG-b-PCL:

Methoxypolyethyleneglycol-block-polycaprolactone

MTS:

3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium

SASCA-A:

Same single cell analysis in the drug accumulation mode

PBS:

Phosphate buffered saline

PCL5:

MePEG17-b-PCL5

PCL200:

MePEG114-b-PCL200

PEN/STR:

Penicillin/streptomycin

P-gp:

Permeability-glycoprotein

SD:

Standard deviation

Tryp/EDTA:

Trypsin-ethylenediaminetetraacetic acid

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Acknowledgment

Financial support from the Natural Science and Engineering Research Council (NSERC) is acknowledged. The authors are grateful to Dr. Wilfred D. Stein for advice in curve-fitting used in the single-cell kinetic study. They thank Yuchun Chen for her assistance in chip design, experimental advice, and curve fitting analysis, and Gagandeep Kaur for her assistance in data processing.

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Authors and Affiliations

  1. Department of Chemistry, Simon Fraser University, 8888 University Drive, Burnaby, BC, V5A 1S6, Canada

    Avid Khamenehfar & Paul C. H. Li

  2. Faculty of Pharmaceutical Sciences, The University of British Columbia, 2146 East Mall, Vancouver, BC, V6T 1Z3, Canada

    Chung Ping Leon Wan, Kevin Letchford & Helen M. Burt

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  1. Avid Khamenehfar
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  2. Chung Ping Leon Wan
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  3. Paul C. H. Li
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Correspondence to Paul C. H. Li.

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Khamenehfar, A., Wan, C.P.L., Li, P.C.H. et al. Same-single-cell analysis using the microfluidic biochip to reveal drug accumulation enhancement by an amphiphilic diblock copolymer drug formulation. Anal Bioanal Chem 406, 7071–7083 (2014). https://doi.org/10.1007/s00216-014-8151-7

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  • DOI: https://doi.org/10.1007/s00216-014-8151-7

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