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DNA Polyplexes of a Phosphorylcholine-Based Zwitterionic Polymer for Gene Delivery

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Abstract

Purpose

We tested polyplexes of a diblock polymer containing a pH-responsive, endosomolytic core (dimethylaminoethyl methacrylate and butyl methacrylate; DB) and a zwitterionic Poly (methacryloyloxyethyl phosphorylcholine) (PMPC) corona for the delivery of plasmid DNA (pDNA) to glioblastoma cells.

Methods

We studied the physicochemical characteristics of the DNA polyplexes such as particle hydrodynamic diameter and surface potential. Cytocompatibility of free PMPC-DB polymer and pDNA polyplexes with U-87MG and U-138MG glioma cell lines were evaluated using the ATP assay. The transfection activity of luciferase pDNA polyplexes was measured using a standard luciferase assay. Anti-proliferative, apoptotic, and cell migration inhibitory activities of PMPC-DB/Interferon-beta (IFN-β1) pDNA polyplexes were examined using ATP assay, flow cytometry, and wound closure assay, respectively.

Results

PMPC-DB copolymer condensed pDNA into nanosized polyplexes. DNA polyplexes showed particle diameters ranging from ca. 100–150 nm with narrow polydispersity indices and near electroneutral zeta potential values. PMPC-DB/Luciferase pDNA polyplexes were safe and showed an 18-fold increase in luciferase expression compared to the gold standard PEI polyplexes in U-87MG cells. PMPC-DB/IFN-β1 polyplexes induced apoptosis, demonstrated anti-proliferative effects, and retarded cell migration in glioblastoma cells.

Conclusion

The results described herein should guide the future optimization of PMPC-DB/DNA delivery systems for in vivo studies.

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Abbreviations

DLS:

Dynamic light scattering

DMAEMA-BMA:

Dimethylaminoethyl methacrylate and butyl methacrylate

GFP:

Green fluorescence protein

IFN-β1:

Interferon beta-1

Luc:

Luciferase

PC:

Phosphatidylcholine

pDNA:

Plasmid deoxyribonucleic acid

PEG:

Polyethylene glycol

PMPC:

Poly (methacryloyloxyethyl phosphorylcholine)

PMPC-DB:

Poly (methacryloyloxyethyl phosphorylcholine) conjugated with DMAEMA-BMA

siRNA:

Small interfering ribonucleic acid

U-138MG:

Uppsala 138 malignant glioma

U-87MG:

Uppsala 87 malignant glioma

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Acknowledgments and Disclosures

The study was supported using start-up funds for the Manickam laboratory from the School of Pharmacy and a Faculty Development Fund award to the PI from Duquesne University (DU). The authors thank Mr. Suyang Wu (DU) for providing Annexin V AF647 and 7-AAD dyes for the flow cytometry studies. The authors express their deep appreciation to Ms. Manisha Chandwani, Ms. Yashika Kamte, and Dr. Lauren O’Donnell (DU) for flow cytometry support. The authors are thankful to Ms. Akshita Bhatt and Dr. Jane Cavanaugh (DU) for fluorescence microscopy support. The authors are thankful to Drs. Joel Gillespie (University of Pittsburgh) and Jelena Janjic (DU) for allowing the use of Malvern ZS90 Zetasizer for the DLS studies. The authors declare no conflicts of interest.

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Authors and Affiliations

  1. Graduate School of Pharmaceutical Sciences, School of Pharmacy, Duquesne University, 600 Forbes Avenue, Pittsburgh, PA, 15282, USA

    Kandarp M. Dave, Linjiang Han, Lindsay Kadlecik & Devika S Manickam

  2. Department of Biomedical Engineering, Vanderbilt University, Nashville, TN, 37232, USA

    Meredith A. Jackson & Craig L. Duvall

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  1. Kandarp M. Dave
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  2. Linjiang Han
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Contributions

K.M.D. and D.S.M. designed the research. M.A.J. and C.L.D. synthesized the PMPC-DB polymer. K.M.D., L.H, and L.K. performed polyplex characterization, polymer cytotoxicity, and luciferase expression studies. K.M.D. and L.K. performed IFN-β1 cell viability studies. K.M.D. performed DOE, flow cytometry, GFP expression, and scratch assays. K.M.D. and D.S.M. analyzed and compiled data. K.M.D. and D.S.M. wrote the manuscript. All authors reviewed the manuscript.

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Correspondence to Devika S Manickam.

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Dave, K.M., Han, L., Jackson, M.A. et al. DNA Polyplexes of a Phosphorylcholine-Based Zwitterionic Polymer for Gene Delivery. Pharm Res 37, 176 (2020). https://doi.org/10.1007/s11095-020-02899-5

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  • DOI: https://doi.org/10.1007/s11095-020-02899-5

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