Delivery of antisense oligodeoxyribonucleotide lipopolyplex nanoparticles assembled by microfluidic hydrodynamic focusing

Chee Guan Koh; Xulang Zhang; Shujun Liu; Sharon Golan; Bo Yu; Xiaojuan Yang; Jingjiao Guan; Yan Jin; Yeshayahu Talmon; Natarajan Muthusamy; Kenneth K. Chan; John C. Byrd; Robert J. Lee; Guido Marcucci; L. James Lee

doi:10.1016/j.jconrel.2009.08.019

Delivery of antisense oligodeoxyribonucleotide lipopolyplex nanoparticles assembled by microfluidic hydrodynamic focusing

Chee Guan Koh, Xulang Zhang, Shujun Liu, Sharon Golan, Bo Yu, Xiaojuan Yang, Jingjiao Guan, Yan Jin, Yeshayahu Talmon, Natarajan Muthusamy, Kenneth K. Chan, John C. Byrd, Robert J. Lee, Guido Marcucci, L. James Lee

Research output: Contribution to journal › Article › peer-review

77 Scopus citations

Abstract

A multi-inlet microfluidic hydrodynamic focusing (MF) system to prepare lipopolyplex (LP) containing Bcl-2 antisense deoxyoligonucleotide (ODN) was developed and evaluated. The lipopolyplex nanoparticles consist of ODN:protamine:lipids (1:0.3:12.5 wt/wt ratio) and the lipids included DC-Chol:egg PC:PEG-DSPE (40:58:2 mol/mol%). Using K562 human erythroleukemia cells, which contain an abundance of Bcl-2 and overexpression of transferrin receptors (TfR), and G3139 (oblimerson sodium or Genasense^TM) as a model cell line and drug, respectively, the Bcl-2 down-regulation at the mRNA and protein levels as well as cellular uptake and apoptosis was compared between the conventional bulk mixing (BM) method and the MF method. The lipopolyplex size and surface charge were characterized by dynamic light scattering (DLS) and zeta potential (ζ) measurement, respectively, while the ODN encapsulation efficiency was determined by gel electrophoresis. Cryogenic transmission electron microscopy (Cryo-TEM) was used to determine the morphology of LPs. Our results demonstrated that MF produced LP nanoparticles had similar structures but smaller size and size distribution compared to BM LP nanoparticles. MF LP nanoparticles had higher level of Bcl-2 antisense uptake and showed more efficient down-regulation of Bcl-2 protein level than BM LP nanoparticles.

Original language	English (US)
Pages (from-to)	62-69
Number of pages	8
Journal	Journal of Controlled Release
Volume	141
Issue number	1
DOIs	https://doi.org/10.1016/j.jconrel.2009.08.019
State	Published - Jan 4 2010
Externally published	Yes

Bibliographical note

Funding Information:
We thank Le-Nguyen Huynh and Jiuxia Pang for helpful discussion on real-time RT-PCR and Western blots. We are grateful to NSF NSEC-CANPBD (Award No. EEC-0425626 ) and NIH for financial support ( R01: CA135243 and R21: 131832 ).

Keywords

Antisense ODN
Bcl-2 down-regulation
G3139
Lipopolyplex
Liposome
Microfluidics
Nanoparticle

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10.1016/j.jconrel.2009.08.019

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Koh, C. G., Zhang, X., Liu, S., Golan, S., Yu, B., Yang, X., Guan, J., Jin, Y., Talmon, Y., Muthusamy, N., Chan, K. K., Byrd, J. C., Lee, R. J., Marcucci, G., & Lee, L. J. (2010). Delivery of antisense oligodeoxyribonucleotide lipopolyplex nanoparticles assembled by microfluidic hydrodynamic focusing. Journal of Controlled Release, 141(1), 62-69. https://doi.org/10.1016/j.jconrel.2009.08.019

Koh, CG, Zhang, X, Liu, S, Golan, S, Yu, B, Yang, X, Guan, J, Jin, Y, Talmon, Y, Muthusamy, N, Chan, KK, Byrd, JC, Lee, RJ, Marcucci, G & Lee, LJ 2010, 'Delivery of antisense oligodeoxyribonucleotide lipopolyplex nanoparticles assembled by microfluidic hydrodynamic focusing', Journal of Controlled Release, vol. 141, no. 1, pp. 62-69. https://doi.org/10.1016/j.jconrel.2009.08.019

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abstract = "A multi-inlet microfluidic hydrodynamic focusing (MF) system to prepare lipopolyplex (LP) containing Bcl-2 antisense deoxyoligonucleotide (ODN) was developed and evaluated. The lipopolyplex nanoparticles consist of ODN:protamine:lipids (1:0.3:12.5 wt/wt ratio) and the lipids included DC-Chol:egg PC:PEG-DSPE (40:58:2 mol/mol%). Using K562 human erythroleukemia cells, which contain an abundance of Bcl-2 and overexpression of transferrin receptors (TfR), and G3139 (oblimerson sodium or GenasenseTM) as a model cell line and drug, respectively, the Bcl-2 down-regulation at the mRNA and protein levels as well as cellular uptake and apoptosis was compared between the conventional bulk mixing (BM) method and the MF method. The lipopolyplex size and surface charge were characterized by dynamic light scattering (DLS) and zeta potential (ζ) measurement, respectively, while the ODN encapsulation efficiency was determined by gel electrophoresis. Cryogenic transmission electron microscopy (Cryo-TEM) was used to determine the morphology of LPs. Our results demonstrated that MF produced LP nanoparticles had similar structures but smaller size and size distribution compared to BM LP nanoparticles. MF LP nanoparticles had higher level of Bcl-2 antisense uptake and showed more efficient down-regulation of Bcl-2 protein level than BM LP nanoparticles.",

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T1 - Delivery of antisense oligodeoxyribonucleotide lipopolyplex nanoparticles assembled by microfluidic hydrodynamic focusing

AU - Koh, Chee Guan

AU - Zhang, Xulang

AU - Liu, Shujun

AU - Golan, Sharon

AU - Yu, Bo

AU - Yang, Xiaojuan

AU - Guan, Jingjiao

AU - Jin, Yan

AU - Talmon, Yeshayahu

AU - Muthusamy, Natarajan

AU - Chan, Kenneth K.

AU - Byrd, John C.

AU - Lee, Robert J.

AU - Marcucci, Guido

AU - Lee, L. James

N1 - Funding Information: We thank Le-Nguyen Huynh and Jiuxia Pang for helpful discussion on real-time RT-PCR and Western blots. We are grateful to NSF NSEC-CANPBD (Award No. EEC-0425626 ) and NIH for financial support ( R01: CA135243 and R21: 131832 ).

PY - 2010/1/4

Y1 - 2010/1/4

N2 - A multi-inlet microfluidic hydrodynamic focusing (MF) system to prepare lipopolyplex (LP) containing Bcl-2 antisense deoxyoligonucleotide (ODN) was developed and evaluated. The lipopolyplex nanoparticles consist of ODN:protamine:lipids (1:0.3:12.5 wt/wt ratio) and the lipids included DC-Chol:egg PC:PEG-DSPE (40:58:2 mol/mol%). Using K562 human erythroleukemia cells, which contain an abundance of Bcl-2 and overexpression of transferrin receptors (TfR), and G3139 (oblimerson sodium or GenasenseTM) as a model cell line and drug, respectively, the Bcl-2 down-regulation at the mRNA and protein levels as well as cellular uptake and apoptosis was compared between the conventional bulk mixing (BM) method and the MF method. The lipopolyplex size and surface charge were characterized by dynamic light scattering (DLS) and zeta potential (ζ) measurement, respectively, while the ODN encapsulation efficiency was determined by gel electrophoresis. Cryogenic transmission electron microscopy (Cryo-TEM) was used to determine the morphology of LPs. Our results demonstrated that MF produced LP nanoparticles had similar structures but smaller size and size distribution compared to BM LP nanoparticles. MF LP nanoparticles had higher level of Bcl-2 antisense uptake and showed more efficient down-regulation of Bcl-2 protein level than BM LP nanoparticles.

AB - A multi-inlet microfluidic hydrodynamic focusing (MF) system to prepare lipopolyplex (LP) containing Bcl-2 antisense deoxyoligonucleotide (ODN) was developed and evaluated. The lipopolyplex nanoparticles consist of ODN:protamine:lipids (1:0.3:12.5 wt/wt ratio) and the lipids included DC-Chol:egg PC:PEG-DSPE (40:58:2 mol/mol%). Using K562 human erythroleukemia cells, which contain an abundance of Bcl-2 and overexpression of transferrin receptors (TfR), and G3139 (oblimerson sodium or GenasenseTM) as a model cell line and drug, respectively, the Bcl-2 down-regulation at the mRNA and protein levels as well as cellular uptake and apoptosis was compared between the conventional bulk mixing (BM) method and the MF method. The lipopolyplex size and surface charge were characterized by dynamic light scattering (DLS) and zeta potential (ζ) measurement, respectively, while the ODN encapsulation efficiency was determined by gel electrophoresis. Cryogenic transmission electron microscopy (Cryo-TEM) was used to determine the morphology of LPs. Our results demonstrated that MF produced LP nanoparticles had similar structures but smaller size and size distribution compared to BM LP nanoparticles. MF LP nanoparticles had higher level of Bcl-2 antisense uptake and showed more efficient down-regulation of Bcl-2 protein level than BM LP nanoparticles.

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KW - Bcl-2 down-regulation

KW - G3139

KW - Lipopolyplex

KW - Liposome

KW - Microfluidics

KW - Nanoparticle

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Delivery of antisense oligodeoxyribonucleotide lipopolyplex nanoparticles assembled by microfluidic hydrodynamic focusing

Abstract

Bibliographical note

Keywords

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