Polyglutamic Acid Grafted Polyethylene Glycol@Calcium Carbonate Based Shielding System for Improving Polyethyleneimine Gene Transfection Efficiency †

doi:10.7503/cjcu20190637

Abstract

Abstract:

Cationic polymers, especially polyethyleneimine(PEI), have received much attention in the field of non-viral vectors, for their better biological safety and high efficiency gene transfection. However, their higher positive charge density will produce greater cytotoxicity, and also, easily forming larger particles with protein, which limits their further usage in vivo. In this work, we designed a shielding system based on polyglutamic acid-polyethylene glycol@calcium carbonate(PPG@CaCO₃) for shielding polyethyleneimine(PEI). On one hand, PGA-PEG(PPG) can reduce the cytotoxicity caused by PEI, and is conducive to applied in vivo. On the other hand, CaCO₃ effectively offset the decreased transfection efficiency that caused by PPG, and even enhanced cell transfection efficiency of PEI. Compared with the polyglutamic acid-polyethylene glycol @calcium phosphate[PPG@Ca₃(PO₄)₂], CaCO₃ releases carbon dioxide gas in acidic environment, which is a key factor to improve cell transfection efficiency. In vivo circulation experiments in mice show that the shielding system based on PPG@CaCO₃ can effectively enhance long circulation. Therefore, the shielding system plays an important role in promoting the in vivo application of cationic gene vectors.

Key words: Polyethyleneimine, Gene transfection, Shielding system, Polyglutamic acid grafted polyethylene glycol@CaCO₃

CLC Number:

O631.1 ⁺1

TrendMD:

GUO Zhaopei,LIN Lin,CHEN Jie,TIAN Huayu,CHEN Xuesi. Polyglutamic Acid Grafted Polyethylene Glycol@Calcium Carbonate Based Shielding System for Improving Polyethyleneimine Gene Transfection Efficiency ^†[J]. Chem. J. Chinese Universities, 2020, 41(2): 235.

Figures/Tables 7

Scheme 1 Preparation of PPG@CaCO3(A) and PPG@Ca3(PO4)2(B)

Fig.1 DLS(A), zeta potential(B) and SEM images(C, D) of PPG@CaCO3(a, C) and PPG@Ca3(PO4)2(b, D)

Fig.2 DLS(A), zeta potentials(B) and SEM images(C—E) of DNA/PEI-25000(a, C), DNA/PEI-25000/PPG@CaCO3(b, D) and DNA/PEI-25000/PPG@Ca3(PO4)2(c, E)

Fig.3 Relative HeLa cell viability of various carrier concentrations (B) a. PEI-1800; b. PEI-1800/PPG@CaCO3; c. PEI-1800/PPG@Ca3(PO4)2.

Fig.4 Transfection efficiency of DNA/PEI, DNA/PEI/PPG, DNA/PEI/PPG@CaCO3, and DNA/PEI/PPG@Ca3(PO4)2 at various mass ratios in HeLa(A, B), CHO(C) and MCF-7(D) cells (A, C, D) a. DNA/PEI-25000; b. DNA/PEI-25000/PPG; c. DNA/PEI-25000/PPG@CaCO3; d. DNA/PEI-25000/PPG@Ca3(PO4)2.(B) a. DNA/PEI-1800; b. DNA/PEI-1800/PPG; c. DNA/PEI-1800/PPG@CaCO3.

Fig.5 Flow cytometric analysis of cell endocytosis efficiency(A) and laser scanning confocal microscope observing particles intracellular transport(B) in HeLa cells a. Control; b. DNA/PEI-25000; c. DNA/PEI-25000/PPG; d. DNA/PEI-25000/PPG@CaCO3; e. DNA/PEI-25000/PPG@Ca3(PO4)2.

Fig.6 Pharmacokinetics of Cy5-DNA, Cy5-DNA/PEI, Cy5-DNA/PEI/PPG, Cy5-DNA/PEI/PPG@CaCO3 and Cy5-DNA/PEI/PPG@Ca3(PO4)2(A) and their half-life in blood(B) a. DNA; b. DNA/PEI-25000; c. DNA/PEI-25000/PPG; d. DNA/PEI-25000/PPG@CaCO3; e. DNA/PEI-25000/PPG@Ca3(PO4)2.

References 21

[1]	Shi B. Y., Zheng M., Tao W., Chung R., Jin D. Y., Ghaffari D., Farokhzad O. C., Biomacromolecules, 2017,18(8), 2231— 2246 doi: 10.1021/acs.biomac.7b00803 URL
[2]	Wang H. X., Li M. Q., Lee C. M., Chakraborty S., Kim H. W., Bao G., Leong K. W., Chem. Rev., 2017,117(15), 9874— 9906 doi: 10.1021/acs.chemrev.6b00799 URL
[3]	Chen J., Li X. Z., Tian H. Y., Zhu X. J., Chen X. S., Chem. J. Chinese Universities, 2015,36(11), 2148— 2156
	( 陈杰, 李小舟, 田华雨, 朱筱娟, 陈学思 . 高等学校化学学报, 2015,36(11), 2148— 2156)
[4]	Tian H. Y., Chen J., Chen X. S., Small, 2013,9(12), 2034— 2044 doi: 10.1002/smll.201202485 URL
[5]	Chen J., Jiao Z. X., Lin L., Guo Z. P., Xu C. N., Li Y. H., Tian H. Y., Chen X. S., Chinese J. Polym. Sci., 2015,33(6), 830— 837 doi: 10.1007/s10118-015-1632-0 URL
[6]	Keles E., Song Y., Du D., Dong W. J., Lin Y. H., Biomater. Sci., 2016,4(9), 1291— 1309 doi: 10.1039/C6BM00441E URL
[7]	Boussif O Lezoualc'h F., Zanta M. A., Mergny M. D., Scherman D., Demeneix B., Behr J. P., ., Proc. Natl. Acad. Sci. USA, 1995,92(16), 7297— 7301 doi: 10.1073/pnas.92.16.7297 URL
[8]	Fichter K. M., Ingle N. P., McLendon P. M., Reineke T. M., ACS Nano, 2013,7(1), 347— 364 doi: 10.1021/nn304218q URL
[9]	Pack D. W., Hoffman A. S., Pun S., Stayton P. S., Nat. Rev. Drug Discovery, 2005,4(7), 581— 593 doi: 10.1038/nrd1775 URL
[10]	Guo Z. P., Tian H. Y., Xia J. L., Chen J., Lin L., Chen X. S., Sci. China:Chem., 2010,53(12), 2490— 2496 doi: 10.1007/s11426-010-4144-3 URL
[11]	Tian H. Y., Guo Z. P., Chen J., Lin L., Xia J. L., Dong X., Chen X. S., Adv. Healthcare Mater., 2012,1(3), 337— 341 doi: 10.1002/adhm.201200033 URL
[12]	Lin L., Guo Z. P., Chen J., Tian H. Y., Chen X. S., Acta Polym. Sin., 2017, ( 2), 321— 328
	( 林琳, 郭兆培, 陈杰, 田华雨, 陈学思 . 高分子学报, 2017, ( 2), 321— 328)
[13]	Guo Z. P., Tian H. Y., Lin L., Chen J., He C. L., Tang Z. H., Chen X. S., Macromol. Biosci., 2014,14(10), 1406— 1414 doi: 10.1002/mabi.201400044 URL
[14]	Chen J., Tian H. Y., Guo Z. P., Xia J. L., Kano A., Maruyama A., Jing X. B., Chen X. S., Macromol. Biosci., 2009,9(12), 1247— 1253 doi: 10.1002/mabi.v9:12 URL
[15]	Guo Z. P., Lin L., Chen J., Zhou X. Z., Chan H. F., Chen X. S., Tian H. Y., Chen M. W., Biomater. Sci., 2018,6(11), 3053— 3062 doi: 10.1039/C8BM00503F URL
[16]	Tian H. Y., Guo Z. P., Lin L., Jiao Z. X., Chen J., Gao S. Q., Zhu X. J., Chen X. S., J. Controlled Release, 2014,174, 117— 125 doi: 10.1016/j.jconrel.2013.11.008 URL
[17]	Guo Z. P., Chen J., Lin L., Guan X. W., Sun P. J., Chen M. W., Tian H. Y., Chen X. S., ACS Appl. Mater. Interfaces, 2017,9(18), 15297— 15306 doi: 10.1021/acsami.7b02734 URL
[18]	Fang H. P., Guo Z. P., Lin L., Chen J., Sun P. J., Wu J. Y., Xu C. N., Tian H. Y., Chen X. S., J. Am. Chem. Soc., 2018,140(38), 11992— 12000 doi: 10.1021/jacs.8b05341 URL
[19]	Cho E. C., Xie J. W., Wurm P. A., Xia Y. N., Nano Lett., 2009,9(3), 1080— 1084 doi: 10.1021/nl803487r URL
[20]	Hatakeyama H., Akita H., Harashima H ., Adv. Drug Delivery Rev., 2011,63(3), 152— 160 doi: 10.1016/j.addr.2010.09.001 URL
[21]	Yu H. Y., Tang Z. H., Li M. Q., Song W. T., Zhang D. W., Zhang Y., Yang Y., Sun H., Deng M. X., Chen X. S., J. Biomed. Nanotechnol., 2016,12(1), 69— 78 doi: 10.1166/jbn.2016.2152 URL