Synthesis of Cyclic Core Dendritic Polymer and Its Usage as a Vector for Transferring Foreign DNA into Human Cells

DendriticpoIymer(ordendrimer)isanewkindofsyntheticpolymer.Dendrimershaveuniquecharactersandsurfaceproperties,thustheyhaveattractedmoreandmoreattCntionofchemistsinrecentyears""'.Dendrimersseemtobepotentiallyusefulmaterialsinpharmacologyandbiology,andtheuseofdendrimerasvectorsforgenetransferisanewattemPt4"".IncomParisonwithotherclassicalpolymersandoligomers,dendrimershavesomeuniquecharactersincludingmonodispersedandsphericalstructurewithregulardendriticbranching.Avarietyofbiologicallyactivemat…     

聚酰胺-胺型树枝状大分子及其衍生物在基因传递中的应用

基因治疗通过基因载体将治病基因导入病患的特异细胞以治疗心血管、神经系统疾病和癌症等。寻找安全高效的非病毒基因载体一直是基因治疗以及生物材料领域中的前沿课题。聚酰胺-胺型(PAMAM)树枝状高分子作为一类三维的、结构高度有序的新型载体,由于具有安全性好、易于修饰、携带外源基因容量大等特点,已经引起了广泛的关注。但是另一方面,合成步骤相对繁琐、后期产物纯化困难以及转染效率相对较低等问题限制了这类载体的进一步发展。本文结合本课题组的研究情况,针对如何提高PAMAM的转染效率以及增强其基因传递的靶向性等相关问题,对近几年在PAMAM树枝状分子修饰改性方面所做的一些有意义的工作进行了综述,并对前景进行了展望。     

In Vitro Drug and Gene Delivery Using Random Cationic Copolymers Forming Stable and pH‐Sensitive Polymersomes

Stimuli‐sensitive polymeric vesicles or polymersomes as self‐assembled colloidal nanocarriers have received paramount importance for their integral role as delivery system for therapeutics and biotherapeutics. This work describes spontaneous polymersome formation at pH 7, as evidenced by surface tension, steady state fluorescence, dynamic light scattering, and microscopic studies, by three hydrophilic random cationic copolymers synthesized using N ,N‐(dimethylamino)ethyl methacrylate (DMAEM) and methoxy poly(ethylene glycol) monomethacrylate in different mole ratios. The results suggest that methoxy poly(ethylene glycol) chains constitute the bilayer membrane of the polymersomes and DMAEM projects toward water constituting the positively charged surface. The polymersomes have been observed to release their encapsulated guest at acidic pH as a result of transformation into polymeric micelles. All these highly biocompatible cationic polymers show successful gene transfection ability as nonviral vector on human cell line with different potential. Thus these polymers prove their utility as a potential delivery system for hydrophilic model drug as well as genetic material.

     

Dual‐Polymer‐Functionalized Nanoscale Graphene Oxide as a Highly Effective Gene Transfection Agent for Insect Cells with Cell‐Type‐Dependent Cellular Uptake Mechanisms

Efficient and safe gene transfection carriers, especially for hard‐to‐transfect cells, are urgently demanded in basic biological research and gene therapy applications. Many insect cell lines widely used in molecular cell biology exhibit relatively low transfection efficiencies when treated by conventional non‐viral agents. Herein, we develop a novel gene delivery vector by coating graphene oxide (GO) with both polyethylene glycol (PEG) and polyethylenimine (PEI), obtaining a dual‐polymer‐functionalized nanoscale GO (nGO‐PEG‐PEI) to transfect insect cells. While exhibiting remarkably reduced cytotoxicity compared with PEI, nGO‐PEG‐PEI, when used as the plasmid DNA transfection agent to treat Drosophila S2 cells, offers ≈7‐fold and ≈2.5‐fold higher efficiency compared with those achieved by using bare PEI and Lipofectamine 2000, a widely used commercial transfection agent, respectively. Interestingly, the advantages of nGO‐PEG‐PEI are even more dramatic when transfecting cells with lower‐quality linearized DNA. It is revealed that nGO‐PEG‐PEI/pDNA complexes enter insect cells via a unique pathway working even at a low temperature, rather different from their entry into mammalian adherent cells. Our results encourage the development of nano‐GO‐based gene carriers to treat special types of hard‐to‐transfect cells (e.g., insect cells), and indicate that nanomaterials would enter cells by cell‐type‐dependent mechanisms, which merit significantly more future attentions.     

Bis-Thioether-Containing Lipid Chains in Cationic Amphiphiles: Physicochemical Properties and Applications in Gene Delivery

Cationic amphiphiles featuring two thioether functions in each lipid chain of bicatenar cationic amphiphiles are reported here for the first time. The physicochemical properties and transfection abilities of these new amphiphiles were compared with those of already reported analogues featuring either (i) saturated, (ii) unsaturated or (iii) mono-thioether containing lipid chains. The homogeneity of the series of new compounds allowed to clearly underscore the effect of bis-thioether containing lipid chains. This study shows that besides previous strategies based on unsaturation or ramification, the incorporation of two thioether functions per lipid chain constitutes an original complementary alternative to tune the supramolecular properties of amphiphilic compounds. The potential of this strategy was evaluated in the context of gene delivery and report that two cationic amphiphiles (i. e. 4 a and 4 b) can be proposed as new efficient transfection reagents.     

Poly(2-oxazoline)-Based Polyplexes as a PEG-Free Plasmid DNA Delivery Platform

The present study expands the versatility of cationic poly(2-oxazoline) (POx) copolymers as a polyethylene glycol (PEG)-free platform for gene delivery to immune cells, such as monocytes and macrophages. Several block copolymers are developed by varying nonionic hydrophilic blocks (poly(2-methyl-2-oxazoline) (pMeOx) or poly(2-ethyl-2-oxazoline) (pEtOx), cationic blocks, and an optional hydrophobic block (poly(2-isopropyl-2-oxazoline) (iPrOx). The cationic blocks are produced by side chain modification of 2-methoxy-carboxyethyl-2-oxazoline (MestOx) block precursor with diethylenetriamine (DET) or tris(2-aminoethyl)amine (TREN). For the attachment of a targeting ligand, mannose, azide-alkyne cycloaddition click chemistry methods are employed. Of the two cationic side chains, polyplexes made with DET-containing copolymers transfect macrophages significantly better than those made with TREN-based copolymer. Likewise, nontargeted pEtOx-based diblock copolymer is more active in cell transfection than pMeOx-based copolymer. The triblock copolymer with hydrophobic block iPrOx performs poorly compared to the diblock copolymer which lacks this additional block. Surprisingly, attachment of a mannose ligand to either copolymer is inhibitory for transfection. Despite similarities in size and design, mannosylated polyplexes result in lower cell internalization compared to nonmannosylated polyplexes. Thus, PEG-free, nontargeted DET-, and pEtOx-based diblock copolymer outperforms other studied structures in the transfection of macrophages and displays transfection levels comparable to GeneJuice, a commercial nonlipid transfection reagent.