基因治疗产品及其载体的研究进展和挑战

基因治疗是一种通过改变个体的基因表达来治疗疾病的方法，为肿瘤、罕见病及其他难治性疾病提供了全新的治疗策略。基因治疗通过基因编辑技术实现对个体致病基因的改造，同时也依赖基因递送载体改善其在体内的稳定性和靶向性。本综述简述了基因治疗的方式，结合目前的基因治疗产品，介绍了病毒、非病毒基因递送载体和基因编辑技术在基因治疗中的发展概况，并总结归纳了临床实践中基因治疗的安全性问题。     

壳聚糖纳米粒载体的应用研究进展

目的介绍壳聚糖纳米粒载体在药物、基因递送等方面的研究应用进展,为其在新领域的应用提供依据。方法广泛查阅中外文有关文献,整理分析归纳了其中27篇文献内容。结果壳聚糖纳米粒载体在药物和基因递送方面已经有诸多研究应用。结论壳聚糖纳米粒载体是一种有前途的非病毒递送载体,其特性和应用有待进一步探索。     

胍基化非病毒载体研究进展

非病毒载体具有质量可控、携带能力高和免疫原性低等优点,近些年来备受关注。理想的非病毒载体在递送基因的过程中需要克服多重生物学屏障,包括血液屏障、细胞屏障和胞内转运屏障。胍基来自于精氨酸,可以与磷酸基团形成双齿氢键,在DNA压缩、细胞膜跨越和入核等过程中发挥着重要作用。对非病毒载体进行胍基化基团的修饰,有利于克服非病毒载体在递送基因的过程中面临的生理屏障,提高基因的转染效率。本文综述了非病毒载体的胍基化修饰对于提高基因递送效率方面的研究进展,为构建新的高效非病毒载体提供了思路。     

阳离子聚合物基因载体的研究进展

与病毒基因载体相比,非病毒基因载体具有毒性低、免疫原性小、结构改造可实现基因靶向递送、可重复应用等优点。而其中阳离子聚合物能够与基因形成稳定复合物,促进细胞对复合物的内吞,受到越来越广泛的关注。本文主要从修饰方法以及靶向传递等方面介绍了阳离子聚合物在基因传递方面的应用。     

精蛋白在改进非病毒载体基因转染中的应用

非病毒基因载体的出现,为基因治疗提供了低毒、易于大规模制备的载体。但与病毒载体相比,非病毒基因载体的转染效率仍然偏低,阻碍了非病毒基因载体的临床应用。本文旨在探讨精蛋白在改进非病毒基因载体方面的应用,希望通过合理的载体设计与优化,制备出一种高效、低毒的基因载体。     

siRNA新型非病毒载体递送策略新进展

小干扰RNA(siRNA)是一个靶向治疗和精确医学的代表性治疗工具，可通过序列特异性的RNA干扰(RNAi)沉默任何疾病相关基因的表达。然而，它的治疗前景历来受到体内半衰期短、递送困难和安全问题的限制。非病毒载体介导的药物递送已经成为克服这些局限性的一个成功策略，可实现siRNA在体内的有效递送，高效沉默靶基因。目前，已有多种药物处于临床试验中，4种基于siRNA的新型疗法已获得美国FDA的批准，标志着靶向疗法新时代的开始。该文概述了近年来基于siRNA的非病毒载体递送策略的新进展及其应用，并展望了siRNA药物研究的未来发展趋势。     

小干扰RNA非病毒递送载体的研究进展

虽然RNA干扰应用在哺乳动物细胞中的时间较短,但siRNA强效抑制靶基因表达的功能使它成为目前广泛研究的一种药物。siRNA作为新型的基因药物可有效治疗多种恶性疾病,比如癌症、病毒性感染疾病、遗传及代谢紊乱性疾病等。siRNA在体内不稳定,易被核酸酶降解,有触发免疫反应的倾向,因此选择安全稳定的载体将其递送到靶组织靶细胞是siRNA在体内发挥作用的前提。非病毒载体低毒、低免疫反应且结构可控,目前已被广泛使用来解决基因递送过程中遇到的困难。文章主要综述了近年来研究广泛的siRNA非病毒递送载体:脂质体、阳离子聚合物、树枝状大分子、无机纳米材料等。     

小干扰RNA治疗药物载体递送技术研究进展

RNA干扰（RNAi）是一种控制靶基因表达的序列特异性调控机制。体内外研究结果表明,RNAi技术的应用对多种疾病治疗有巨大的前景,其瓶颈在于如何将基于RNAi的药物有效递送至靶细胞或靶组织。常用的载体递送系统可分为病毒载体系统和非病毒载体系统。研究载体递送系统需重点考虑的因素有脱靶效应、递送方式、免疫反应诱导及剂量等。如果这些关键问题能够解决,将大大增加RNAi成为治疗药物的可能性。     

siRNA体内递送研究进展

RNA干扰(RNA interference,RNAi),是一种在动植物中存在的通过双链RNA诱导同源特异性序列转录后基因沉默的过程。虽然小干扰RNA (siRNA) 较单链反义寡核苷酸显示出更好的稳定性与基因沉默效果,但是作为新型的基因治疗药物,靶向递送siRNA是药物进入临床应用最主要的环节,siRNA体内有效作用发挥的关键在于它在体内能否高效递送至靶细胞并与靶基因结合。目前研究主要集中在siRNA的修饰方式与递送载体研究,以提高其体内的稳定性与靶向性。文中主要综述了siRNA的体内靶向递送障碍以及近几年siRNA非病毒递送载体脂质体、阳离子多聚物、纳米粒、胶束等方面的研究进展。     

小分子干扰RNA非病毒纳米载体的设计进展

小分子干扰RNA(small interfering RNA,siRNA)进入临床应用最关键的环节是如何有效通过药物递送系统将siRNA递送到特异性靶细胞或靶组织。非病毒纳米载体递送系统具有低的免疫原性、良好的靶向性和生物相容性,是近年来国内外药剂工作者研究的热点。其相应的研究也必将大大增加siRNA有效递送,使其有进一步成为治疗药物的可能性。本文根据国内外的文献报道,将siRNA非病毒纳米载体的设计进展进行了综述。     

用于肿瘤治疗的小分子干扰RNA非病毒载体研究进展

近年来,小分子干扰RNA(siRNA)作为RNA干扰 (RNAi) 技术的效应分子,已被广泛用于恶性肿瘤的基因治疗领域。欲获得理想的治疗效果,其关键因素是寻找一种安全、高效、稳定、可控的基因载体。非病毒载体具有低毒、低免疫原性、制备简单、目的基因容量大、外源基因随机整合率低且携带基因大小类型不受限制等突出优势,已经成为目前siRNA载体的研究热点。在以往学者的研究基础上,从药剂学的角度,笔者对这些载体在siRNA传递系统中的研究现况做回顾性总结。     

疫苗和基因给药系统的研究进展

简述了当前疫苗和基因给药系统的研究进展，包括注射、口服、鼻腔和透皮等给药系统，并介绍了以转基因植物作为疫苗以及基因给药系统的病毒载体和非病毒载体系统。     

Cationic lipids containing cyclen and ammonium moieties as gene delivery vectors

In this study, two novel cationic lipids containing protonated cyclen and quaternary ammonium moieties were designed and synthesized as non-viral gene delivery vectors. The structures of the two lipids differ in their hydrophobic region (cholesterol or diosgenin). Cationic liposomes were easily prepared from the lipids individually or from the mixtures of each cationic lipid and dioleoylphosphatidylethanolamine. Several studies including DLS, gel retardation assay, and ethidium bromide intercalation assay suggest that these amphiphilic molecules are able to bind and compact DNA into nanometer particles which can be used as non-viral gene delivery agents. Our results from in vitro transfection show that in association with dioleoylphosphatidylethanolamine, two cationic lipids can induce effective gene transfection in human embryonic kidney 293 cells, although the gene transfection efficiencies of two cationic lipids were found to be lower than that of lipofectamine 2000(TM) . Besides, different cytotoxicity was found for two lipoplexes. This study demonstrates that the title cationic lipids have large potential to be efficient non-viral gene vectors.     

Comparative analysis of the methods of drug and protein delivery for the treatment of cancer,genetic diseases and diagnostics

The methods of protein and drug delivery for the treatment of cancer, genetic diseases and diagnostics were summarized. The potential of protein transduction is discussed and the recent developments in the field are reviewed. An overview is provided of the non-viral delivery methods such as liposomes, polymer-based delivery, cell-penetrating peptides, bacterial secretion, cells, virosomes, physical methods including electroporation, microinjection, osmotic lysis, nanoparticles, sonoporation to locally inject therapeutic molecules. The characteristic properties of non-viral vectors and their use for the delivery of therapeutic molecules for the diagnosis and treatment of disorders and to target tumors are also discussed. The potential of the transduced peptides and proteins was used as new therapeutic compounds against infectious diseases, to complement deficiencies in specific genes, to specifically kill tumour cells, for gene therapy. The protein delivery vectors can enhance the transfection at low concentrations and help to develop future gene delivery systems with reduced toxicity. Vitamin B12, folic acid, biotin, and riboflavin are essential in the treatment of cancer. Ultrasound has a potential in the delivery of therapeutic agents. The new developing technologies of drug delivery and targeting offer the possibility to improve the therapeutic possibilities of the existing drugs and to develop novel therapeutics.     

基于聚乙烯亚胺的非病毒基因载体研究进展

如今,越来越多的非病毒基因载体被应用于基因传递中,聚乙烯亚胺（PEI）作为一种重要的阳离子聚合非病毒基因传递载体受到广泛的关注。针对如何克服其生物不可降解性和细胞毒性并进一步提高转染效率的问题,笔者通过文献综合介绍了一些新的策略,并具体对以PEI为基础的新型载体的合成、新型体外转染方法的建立以及体内应用等方面的研究进展进行综述。     

Advances with the use of bio-inspired vectors towards creation of artificial viruses

Importance of the field: In recent years, there has been a great deal of interest in the development of recombinant vectors based on biological motifs with potential applications in gene therapy. Several such vectors have been genetically engineered, resulting in biomacromolecules with new properties that are not present in nature.

Areas covered in this review: This review briefly discusses the advantages and disadvantages of the current state-of-the-art gene delivery systems (viral and non-viral) and then provides an overview on the application of various biological motifs in vector development for gene delivery. Finally, it highlights some of the most advanced bio-inspired vectors that are designed to perform several self-guided functions.

What the reader will gain: This review helps the readers get a better understanding about the history and evolution of bio-inspired fusion vectors with the potential to merge the strengths of both viral and non-viral vectors in order to create efficient, safe and cost-effective gene delivery systems.

Take home message: With the emergence of new technologies such as recombinant bio-inspired vectors, it may not take long before non-viral vectors are observed that are not just safe and tissue-specific, but even more efficient than viral vectors.     

Antisense strategies and non-viral gene therapy for cancer

Effective gene therapy for cancer remains an elusive goal, even after more than a decade of intensive research. There has been, however, tremendous progress in the development of increasingly sophisticated non-viral (or synthetic) delivery vectors for local and systemic administration of nucleic acids. Recent clinical data has also indicated the feasibility of using antisense oligonucleotides to inhibit inappropriately expressed or mutated genes in human cancers. The purpose of this review is to provide an update of the patent literature on the development of non-viral approaches for cancer gene therapy. In particular, patents on lipoplexes and polyplexes for delivery of therapeutic genes and antisense oligonucleotides are reviewed. The diverse range of antisense strategies being developed and recent clinical data are also highlighted.     

Poly(amidoamine) dendrimer complexes as a platform for gene delivery

Introduction: Gene therapy is one of the most effective ways to treat major infectious diseases, cancer and genetic disorders. It is based on several viral and non-viral systems for nucleic acid delivery. The number of clinical trials based on application of non-viral drug and gene delivery systems is rapidly increasing.

Areas covered: This review discusses and summarizes recent advances in poly(amidoamine) dendrimers as effective gene carriers in vitro and in vivo, and their advantages and disadvantages relative to viral vectors and other non-viral systems (liposomes, linear polymers) are considered.

Expert opinion: In this regard, dendrimers are non-immunogenic and have the highest efficiency of transfection among other non-viral systems, and none of the drawbacks characteristic for viral systems. The toxicity of dendrimers both in vitro and in vivo is an important question that has been addressed on many occasions. Several non-toxic and efficient multifunctional dendrimer-based conjugates for gene delivery, along with modifications to improve transfection efficiency while decreasing cytotoxicity, are discussed. Twelve paradigms that affected the development of dendrimer-based gene delivery are described. The conclusion is that dendrimers are promising candidates for gene delivery, but this is just the beginning and further studies are required before using them in human gene therapy.