Journal of Materials Chemistry B
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2017
·Vol. 5
·期 39
·7963-7973
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IF 4.543
Y. N. Zhao,Y. Z. Piao,C. M. Zhang,Y. M. Jiang,A. Liu,S. H. Cui,D. F. Zhi,Y. H. Zhen,S. B. Zhang
Headgroups in cationic lipids play very important roles in determining transfection efficiency and toxicity in gene delivery. To better understand the influence of headgroups on gene delivery, a tri-peptide-based lipid was synthesized, wherein the usual quaternary ammonium was replaced by a tri-peptide. Though both the tri-peptide-based lipid (DAO3) and the quaternary ammonium-based lipid (DDCTMA) successfully mediated gene transfection, DAO3 was superior to DDCTMA in both in vitro and in vivo studies. Following their preparation into liposomes, the particle size, zeta potential, and DNA-binding capacity of the liposomes and lipoplexes were characterized to evaluate the efficiency of DAO3 compared to DDCTMA with regard to gene interactions. The expression of luciferase from pDNA mediated by DAO3 was 2-fold greater than than that with DDCTMA in Hep-2 cells, and DAO3/siRNA lipoplexes could silence about 60% luciferase in A549 cancer cells expressing firefly luciferase. DAO3/Luc-siRNA treatment exhibited 3-fold the efficiency of DDCTMA/Luc-siRNA in terms of in vivo luciferase RNAi with the bare density ratio of 0.54 at 48 h. Furthermore, DAO3 could mediate IGF-1R siRNA to inhibit tumor growth through silencing the expression of the IGF-1R protein, whereas DDCTMA showed nearly no effects. Most importantly, DAO3 had no obvious toxicity in vitro and in vivo, due to the biocompatibility of the peptide headgroups. In conclusion, these results demonstrated that the replacement of the quaternary ammonium headgroup by tri-ornithine may increase transfection efficiency and decrease toxicity.
研究领域:
分子生物学
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基因治疗
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药物递送
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PubMed
高影响因子
Journal of Materials Chemistry B
·
2017
·Vol. 5
·期 7
·1482-1497
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IF 4.543
Xiaobing Chen,Jun Yang,Hong Liang,Qian Jiang,Bowen Ke,Yu Nie
Integrating the advantages of artificial vectors with bioinspired strategies has opened interesting perspectives in the design of gene delivery systems. Herein, disulfide modified, self-assembled lipopeptides with arginine-rich periphery are demonstrated as gene vectors (RLS). It is surprising that these carriers achieved excellent gene transfection efficacy (up to 380-fold higher than PEI) in different cell lines (HeLa and B16 cells) at relatively low N/P (∼10) ratios, compared to the analog lipopeptides without disulfide bonds (RL, N/P 40). As shown from the morphologies observed by transmission electronic microscopy and surface charge detection by dynamic light scattering, the existence of disulfide bonds may influence the configuration/conformation of the RLS assembly with decreased zeta potential (+27.2 mV), compared to that of the analogs, RL (+46.5 mV). Correspondingly, RLS/DNA complexes showed relatively lower zeta potentials than those of RL/DNA complexes at different N/P ratios. Under the transfection conditions, RLS/DNA complexes (N/P 10) showed even less cellular uptake than that of RL/DNA complexes (N/P 40), and no difference was detected in buffering effect and endosomal escape between RLS and RL complexes. Examination by gel electrophoresis and fluorescence resonance energy transfer (FRET) images in cell culture conditions confirmed that RLS lipoplexes could effectively liberate DNA. Thus, the higher gene transfection efficiency of disulfide modified vectors might be mainly attributed to the cleavage of disulfide bonds; the rapid release of DNA and the superiority of the design was closely related to the reductive conditions in the cells.
研究领域:
生物化学
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分子生物学
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药物递送
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DOI
PubMed
高影响因子
JOURNAL OF CONTROLLED RELEASE
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2011
·
IF 7.164
Congxin Li,Tianying Guo,Dezhong Zhou,Yuling Hu,Hao Zhou,Shufang Wang,Jiatong Chen,Zhengpu Zhang
A novel non-viral gene vector based on poly[poly(ethylene glycol) methacrylate] (PMPEG) and l -glutathione (GSH) grafted chitosan (CS) has been fabricated. First, well-defined brush-like PMPEG living polymers with dithioester residues were prepared by the reversible addition–fragmentation chain transfer (RAFT) polymerization and grafted onto the allylchitosan via radical coupling method. Then, the tripeptide GSH was introduced onto the end of PMPEG chain to give a CS–PMPEG–GSH conjugate. In comparison with pristine chitosan, CS–PMPEG–GSH conjugate could not only condense plasmid DNA (pDNA) and prevent the condensed CS–PMPEG–GSH/pDNA nanoparticle self-aggregation, but also increase the binding ability to cell membrane efficiently and improve decondensed ability of pDNA from the nanoparticles in cytoplasm which thus has resulted in the higher transfection efficiency in mouse embryonic fibroblast cells (NIH3T3). In addition, cytotoxicity assays showed that the conjugate is less cytotoxic than CS, and still retain the cationic polyelectrolyte characteristic as chitosan. These results indicate that the non-viral vector is a promising candidate for gene therapy in clinical application.
研究领域:
生物材料学
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分子生物学
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基因治疗
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DOI
PubMed
高影响因子
BIOMACROMOLECULES
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2012
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IF 5.479
Xiaojun Cai,Chunyan Dong,Haiqing Dong,Gangmin Wang,Giovanni M. Pauletti,Xiaojing Pan,Huiyun Wen,Isaac Mehl,Yongyong Li,Donglu Shi
A dual stimulus-responsive mPEG-SS-PLL15-glutaraldehyde star (mPEG-SS-PLL15-star) catiomer is developed and biologically evaluated. The catiomer system combines redox-sensitive removal of an external PEG shell with acid-induced escape from the endosomal compartment. The design rationale for PEG shell removal is to augment intracellular uptake of mPEG-SS-PLL15-star/DNA complexes in the presence of tumor-relevant glutathione (GSH) concentration, while the acid-induced dissociation is to accelerate the release of genetic payload following successful internalization into targeted cells. Size alterations of complexes in the presence of 10 mM GSH suggest stimulus-induced shedding of external PEG layers under redox conditions that intracellularly present in the tumor microenvironment. Dynamic laser light scattering experiments under endosomal pH conditions show rapid destabilization of mPEG-SS-PLL15-star/DNA complexes that is followed by facilitating efficient release of encapsulated DNA, as demonstrated by agarose gel electrophoresis. Biological efficacy assessment using pEGFP-C1 plasmid DNA encoding green fluorescence protein and pGL-3 plasmid DNA encoding luciferase as reporter genes indicate comparable transfection efficiency of 293T cells of the catiomer with a conventional polyethyleneimine (bPEI-25k)-based gene delivery system. These experimental results show that mPEG-SS-PLL15-star represents a promising design for future nonviral gene delivery applications with high DNA binding ability, low cytotoxicity, and high transfection efficiency.
研究领域:
生物医学工程
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分子生物学
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药剂学
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DOI
PubMed
高影响因子
EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY
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2018
·
IF 4.816
Ya-Ping Xiao,Ji Zhang,Yan-Hong Liu,Ju-Hui Zhang,Qing-Ying Yu,Zheng Huang,Xiao-Qi Yu
Fluorinated biomaterials have been reported to have promising features as non-viral gene carriers. In this study, a series of fluorinated polymeric gene carriers were synthesized via Michael addition from low molecular weight polyethyleneimine (PEI) and fluorobenzoic acids (FBAs)-based linking compounds with different numbers of fluorine atoms . The structure-activity relationship (SAR) of these materials was systematically investigated. SAR studies showed that fluorine could screen the positive charge of these polymers. However, this shielding effect of fluorine would endow fluorinated polymers with good balance between DNA condensation and release. In vitro transfection results suggested that these fluorinated polymers could mediate efficient gene delivery. Flow cytometry and confocal microscopy studies demonstrated that more efficient cell uptake could be achieved by fluorinated materials with more fluorine atoms. Cytotoxicity assays showed that these fluorinated materials exhibited very low cytotoxicity even at high mass ratios. This study demonstrates that FBA-based fluorinated biopolymers have the potential for practical application.
研究领域:
生物材料学
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基因治疗
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分子生物学
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DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
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2016
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IF 7.145
Jing Zhao,Ping Huang,Zhiyong Wang,Yan Tan,Xiaohu Hou,Liping Zhang,Cheng-Yi He,Zhi-Ying Chen
Minicircle DNA (mcDNA) is a kind of enhanced nonviral DNA vector with excellent profiles in biosafety and transgene expression. Herein, we reported a novel amphiphilic polymer comprising polyethylenimine(PEI) modified Poly(β-amino ester) PEI–PBAE(C16) for efficient mcDNA delivery in vivo. The synthesized polymer could condense mcDNA into nanoscaled structure and exhibited efficient gene transfection ability without detectable cytotoxicity. Importantly, when injected into mouse intraperitoneally, these PEI–PBAE(C16) nanocomplexes were able to result in high level of trangene expression which lasted at least 72 h. Overall, these results demonstrated the PEI–PBAE(C16) can mediate effective and safe gene delivery in vivo with clinical application potential.
研究领域:
生物化学
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基因治疗
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分子生物学
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DOI
PubMed
高影响因子
Polymer Chemistry
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2017
·Vol. 8
·期 48
·7486-7498
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IF 5.375
Bing Wang,Ping Chen,Ji Zhang,Xiao-Chuan Chen,Yan-Hong Liu,Zheng Huang,Qing-Ying Yu,Ju-Hui Zhang,Wei Zhang,Xi Wei,Xiao-Qi Yu
With the aim to develop a novel multifunctional gene delivery system that may overcome the common barriers of gene transfection, two amphiphilic molecules with tetraphenyl ethylene (TPE) as a hydrophobic moiety and linear PEI-galactose as a hydrophilic part were elaborately designed and synthesized. The molecules could self-assemble to form micelles and exhibited aggregation-induced emission (AIE) property, which may be used for cell imaging. Several assays revealed that the disulfide bond in TSPG may facilitate vector decomposition and DNA release, resulting in higher transfection efficiency and lower cytotoxicity than TCPG and PEI 25 kDa. These vectors showed excellent serum tolerance, and even higher efficiency could be obtained in transfection with the presence of serum. Moreover, the transfection in different cell lines showed that TSPG complex could induce higher gene expression in asialoglycoprotein receptor-rich HepG2 cells than in HeLa cells, which lack such receptors. Galactose competition assay also demonstrated that the galactose moiety could afford the vector with targeting ability toward hepatoma cells. Cellular uptake inhibition assay indicated that the DNA complex entered the cell mainly via caveolae-mediated endocytosis. Results showed that TSPG may serve as a promising reagent for simultaneous gene transfection and imaging with good biocompatibility and cell targeting ability.
研究领域:
生物医学工程
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分子生物学
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药剂学
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DOI
高影响因子
Advanced Healthcare Materials
·
2019
·Vol. 8
·期 8
·1800934
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IF 6.27
Yuan Liu,Dan Chen,Jialin Li,Dengning Xia,Miaorong Yu,Jinsong Tao,Xinxin Zhang,Li Li,Yong Gan
Gene vectors for oral delivery encounter harsh conditions throughout the gastrointestinal tract, and the continuous peristaltic activity can quickly remove the vectors, leading to inefficient intestinal permeation. Therefore, vectors have demanding property requirements, such as stability under various pH and, more importantly, efficient uptake in different intestinal segments. In this study, a functional polymer, cholesterol-grafted poly(β-amino ester) (poly[hexamethylene diacrylate-β-(5-amino-1-pentanol)] (CH-PHP)), is synthesized and electrostatically interacted with plasmid DNA to form a CH-PHP/DNA complex (CPNC). This complex is designed to target the Niemann–Pick C1-like receptor, a cholesterol receptor, to improve oral gene delivery efficacy. With the presence of cholesterol, CH-PHP shows mitigated cytotoxicity, enhanced enzyme resistance, and improved gene condensing ability. CPNC further contributes to ≈43.1- and 2.3-fold increases in luciferase expression in Caco-2 cells compared with PNC and Lipo 2000/DNA complexes, respectively. In addition, the in vivo transfection efficacy of CPNC is ≈4.1-, 2.1-, and 1.6-fold higher than that of Lipo 2000/DNA complexes in rat duodenum, jejunum, and ileum, respectively. Therefore, CPNC may be a promising delivery vector for gene delivery, and using a cholesterol-specific endocytic pathway can be a novel approach to achieve efficient oral gene transfection.
研究领域:
药剂学
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分子生物学
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DOI
PubMed
高影响因子
CHINESE JOURNAL OF POLYMER SCIENCE
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2015
·Vol. 34
·期 1
·94-103
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IF 1.835
Zhao Bin-wei,Zhou Zhuxian,Shen Youqing
Chirality is a key factor in the biological activity of many biomolecules. Poly(L-lysine) (PLL), a polypeptide synthesized from L-lysine, is one of the mostly used cationic polymers for gene delivery. The effect of chirality of polylysine (PL) on its gene delivery remains unknown. Herein, we prepared three polylysines (PLs) with the similar molecular weight but different backbone chiralities including poly(L-lysine) (PLL), poly(D-lysine) (PDL) and poly(DL-lysine) (PDLL). The side chains of each PL were modified with propylene oxide (PO) of different chiralities including (R)PO, (S)PO and (R,S)PO. These PL-POs with distinct chirality in main and side chains could condense pDNA into polyplexes. The polyplexes had approximately the same size, zeta potential and binding ability, but showed distinct gene transfection efficiency. We found that the PLs of L-configuration in the main chain had higher transfection efficiency than that of D or DL configuration due to their faster cellular uptake, while the side chain chirality had no effect on transfection efficiency.
研究领域:
生物化学
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基因治疗
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分子生物学
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DOI
高影响因子
Polymer Chemistry
·
2012
·Vol. 4
·期 1
·156-165
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IF 5.321
Lejiao Jia,Zhenyu Li,Dianrui Zhang,Qiang Zhang,Jingyi Shen,Hejian Guo,Xiaona Tian,Guangpu Liu,Dandan Zheng,Lisi Qi
The chitosan oligosaccharide-based disulfide-containing polyethylenimine derivative PEG-ss-COS-ss-PEI was synthesized and evaluated as a nonviral gene delivery carrier. The structure of the obtained polymers was confirmed by 1H NMR and FTIR. PEG-ss-COS-ss-PEI copolymers could effectively condense DNA into small particles with average diameters less than 120 nm and the zeta potential of +15.7 mV at the N/P ratio of 15/1. Additionally, the resultant polyplexes showed excellent colloidal stability against 150 mM NaCl and had a better buffering capacity of ∼44%, which was more than double the buffering capacity of PEI1.8k (∼20%). In the presence of 10 mM glutathione (GSH), however, polyplexes of PEG-ss-COS-ss-PEI were rapidly unpacked, as revealed by significant increase of particle sizes to over 800 nm. In vitro experiments revealed that the PEG-ss-COS-ss-PEI copolymers not only had much lower cytotoxicity, but also displayed high transfection efficiency as compared to the control branch 25 kDa PEI. This study indicates that a reducibly degradable copolymer PEG-ss-COS-ss-PEI composed of low molecular weight PEI, chitosan oligosaccharide and PEG via disulfide-containing linkages can be a promising gene delivery carrier.
研究领域:
生物材料学
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基因治疗
·
药剂学
✓ 已验证
DOI
高影响因子
LANGMUIR
·
2018
·
IF 3.789
Shengran Li,Binggang Chen,Yangchun Qu,Xinxin Yan,Wenliang Wang,Xiaojing Ma,Bo Wang,Sanrong Liu,Xifei Yu
As one of the most promising therapeutic methods, gene therapy has been playing a more and more important role in treating disease due to its ultra-high therapy efficiency. Even if nonviral gene vectors represented by polycation, liposomal, dendrimers, and zwitterionic materials have made great progress in gene complexation, low immunogenicity, and biocompatibility, intracellular gene release with low toxicity is effectively still a bottleneck restricting the clinical application of gene therapy. We designed and synthesized a reactive oxygen species (ROS)-responsive dendrimer poly(amido amine)-N-(4-boronobenzyl)-N,N-diethyl-2-(propionyloxy)ethan-1-aminium (PAMAM-(B-DEAEP)16) as a gene vector whose potential can vary from positive to negative under the elevated ROS (H2O2) in cancerous cells. Dynamic light scattering results showed that the zeta potential of PAMAM-(B-DEAEP)16 decreased from +12.3 to −5 mV under 80 mM H2O2 in PBS buffer. The 1H NMR results demonstrated that the intermediate status of PAMAM-(B-DEAEP)16 was zwitterionic in ∼6 h because it consisted of the positive quaternary ammonium and negative carboxylic acid simultaneously before the ester bond was completely hydrolyzed. Gel retardation assay showed that PAMAM-(B-DEAEP)16 can condense DNA at above N/P = 1; then, PAMAM-(B-DEAEP)16 transfers to zwitterionic, which begins to continuously release DNA with the decrease in the positive charges and increase in the negative charges, and finally to negatively charged poly(amido amine)-propionic acid (PAMAM-PAc16) in the 80 mM H2O2. Fluorescence-labeled Cy-5 DNA indicated that PAMAM-(B-DEAEP)16 can enter into the cell completely in ∼4 h. The results showed that this compound we designed exhibited higher gene transfection efficiency and lower cytotoxicity than commercial PEI. This is the first time that the positively charged dendrimer was transferred to zwitterionic dendrimer under the stimuli of H2O2 and was successfully applied to gene delivery. Unlike all of the previous reports, we did not seek a compromise between the high gene transfection and low toxicity but find a new avenue to make the gene carrier not only have higher gene transfection efficiency but also exhibit lower toxicity by introducing stimuli-sensitive groups into the positively charged dendrimer to make it capable of adjusting the charge property according to the microenvironment. This study not only provides a good method to design materials for gene delivery but also opens a new perspective to understand the process of gene delivery.
研究领域:
生物医学工程
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基因治疗
·
材料科学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2012
·
IF 2.453
Dezhong Zhou,Congxin Li,Yuling Hu,Hao Zhou,Jiatong Chen,Zhengpu Zhang,Tianying Guo
Poly( l -lysine) (PLL) has excellent plasmid DNA (pDNA) condensation capacity. However, the relatively high cytotoxicity and low transfection efficiency limit its application as gene delivery vectors. Here, well-defined glycopolymers are synthesized by reversible addition fragmentation transfer polymerization and grafted onto PLL to improve the gene delivery performance. After glycopolymer modification, PLL shows reduced cytotoxicity. By regulating the glycopolymer length and amino group substitution degree, the glycopolymer modified PLL can condense pDNA with proper strength, protect the condensed pDNA from degradation and release them in time. Transfection with NIH3T3 and HepG2 cells shows that the glycopolymer modified PLL has improved transfection efficiencies. The low cytotoxicity, effective pDNA protection and enhanced transfection efficiencies indicate that glycopolymer modification would be an effective strategy to improve the polycation properties for gene delivery.
研究领域:
生物材料学
·
分子生物学
·
基因治疗
✓ 已验证
DOI
PubMed
高影响因子
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