개선된 플라스미드 DNA 전달 효율을 위한 히알루론 아민 코팅 폴리에틸렌이민 기반 전달 시스템

오경연; 장용호; 이은비; 김태호; 김현철; Kyoung-yeon Oh; Yongho Jang; Eunbi Lee; Tae-ho Kim; Hyuncheol Kim

Applied Chemistry for Engineering, Vol.33, No.1, 83-89, February, 2022

DOI10.14478/ace.2021.1103 Export Citation

개선된 플라스미드 DNA 전달 효율을 위한 히알루론 아민 코팅 폴리에틸렌이민 기반 전달 시스템

Polyethyleneimine based Delivery System Coated with Hyaluronate Amine for Improved pDNA Transfection Efficiency

오경연, 장용호, 이은비, 김태호, 김현철^†

서강대학교 화공생명공학과

Kyoung-yeon Oh, Yongho Jang, Eunbi Lee, Tae-ho Kim, and Hyuncheol Kim^†

Department of Chemical and Biomolecular Engineering, Sogang University, Seoul 04107, South Korea

E-mail:

초록

현재 진행 중인 코로나19의 세계적 유행을 기점으로 유전자 전달을 통한 면역 형성에 대한 연구가 활발히 진행되고 있다. 특히 바이러스를 통한 유전자 전달이 부작용이 다수 발견됨에 따라 비바이러스성 유전자 전달체에 대한 요구가 크게 증가하였다. 본 연구에서는 생체적합물질인 히알루론 아민으로 코팅한 폴리에틸렌이민-플라스미드 DNA 복합체를 통한 효율적인 유전자 전달 시스템을 제안했다. 다양한 조성에서 생성된 폴리에틸렌이민-플라스미드 DNA 복합체 (polyplex)와 히알루론 아민으로 코팅한 폴리에틸렌이민-플라스미드 DNA 복합체(polyplex-HA)의 크기 및 플라스미드 DNA 발현 정도를 비교해 각 물질의 최적 비율을 찾아냈고 복합체의 크기 및 제타 전위, 에너지 필터링 투과 전자 현미경(EF-TEM) 이미지를 통해 입자의 특성을 평가했다. 세포 내 전달 및 발현 효율을 형광현미경과 유세포분석기를 통해 상용화 되어있는 유전자 전달체인 lipofectamine과 비교 분석했다. 본 연구에서 제안된 polyplex-HA는 pDNA 뿐만 아니라 다양한 유전물질을 전달할 수 있으며, 전달체에 대한 면역반응이 적어 다회성 투여에 유리하여 미래의 백신 플랫폼의 기반이 될 수 있을 것으로 기대할 수 있다.

Since the pandemic of COVID-19, active investigation to develop immunity to infectious disease by delivering nucleic acids has been proceeded. Particularly, many studies have been conducted on non-viral vector as several vital side-effects which were found on nucleic acid delivery system using viral vectors. In this study, we have developed plasmid DNA (pDNA) loaded-hyaluronic acid derivative (HA) coated-polyethyleneimine (PEI) based polyplex for enhanced nucleic acid delivery efficiency. We have optimized the ratio of pDNA : PEI : HA by measuring size and protein transcription efficiency. The final product, polyplex-HA, was characterized through measuring size, zeta-potential and TEM image. Intracellular uptake and protein transcription efficiency were compared to commercially available transfection reagent, lipofectamine, through fluorescence image and flow cytometry. In conclusion, polyplex-HA presents a novel gene delivery system for efficient and stable protein transcription since it is available for delivering various genetic materials and has less immunoreactivity.

Keywords:Vaccine;Nanoparticles;Plasmid DNA;Polyethyleneimine (PEI)

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[2] Pardi N, Hogan MJ, Porter FW, Weissman D, Nat. Rev. Drug Discov., 17, 261 (2018)

[3] Zhang NN, LI XF, Deng YQ, Zhao H, Huang YJ, Yang G, Qin CF, Cell, 182, 1271 (2020)

[4] Liu MA, Vaccines, 7, 37 (2019)

[5] Nguyen HH, Park J, Park SJ, Lee CS, Hwang S, Shin YB, Ha TH, Kim M, Polymers, 10, 28 (2018)

[6] Roy P, Noad R, Hum. Vaccin., 4, 5 (2008)

[7] Yang ZY, Wyatt LS, Kong WP, Moodie Z, Moss B, Nabel GJ, J. Virol., 77, 799 (2003)

[8] Wylon K, Dolle S, Worm M, Allergy Asthma Clin. Immunol., 12, 1 (2016)

[9] Uzgun S, Nica G, Pfeifer C, Bosinco M, Michaelis K, Lutz JF, Schneider M, Rosenecker J, Rudolph C, Pharm. Res., 28, 2223 (2011)

[10] Webster R, Elliott V, Park BK, Walker D, Hankin M, Taupin P, PEGylated protein drugs: Basic science and clinical applications, 127 (2009).

[11] Smith SA, Selby LI, Johnston AP, Such GK, Bioconjugate Chem., 30, 263 (2018)

[12] Boussif O, Lezoualc'h F, Zanta MA, Mergny MD, Scherman D, Demeneix B, Behr JP, Proc. Nat. Acad. Sci., 92, 7297 (1995)

[13] Yang Y, Jia Y, Xiao Y, Hao Y, Zhang L, Chen X, He J, Zhao Y, Qian Z, ChemphysChem, 19, 2058 (2018)

[14] Benjaminsen RV, Mattebjerg MA, Henriksen JR, Moghimi SM, Andresen TL, Mol. Ther., 21, 149 (2013)

[15] Chen J, Tian H, Kano A, Maruyama A, Chen X, Park TG, J. Control. Release, 152, e134 (2011)

[16] Racine R, Mummert , Molecular Regulation of Endocytosis, 377 (2012).

[17] Akinc A, Thomas M, Klibanov AM, Langer R, J. Gene Med., 7, 657 (2005)