Viral delivery for gene therapy against cell movement in cancer

Viral delivery for cancer gene therapy is a promising approach, where traditional radiotherapy or chemotherapy to limit proliferation and movement of cancer cells has met resistance. Based on the new understanding of the biology of the viral vectors, therapeutic viral vectors for cancer gene therapy have been improved for greater safety and efficacy as well as transitioned from being non-replicating to replication-competent. Traditional oncolytic vectors have focused on eliminating tumor growth, while novel vectors simultaneously target epithelial-to-mesenchymal transition (EMT) in cancer cells, which could further prevent and reverse the aggressive tumor progression. In this review, we highlight the illustrative examples of cancer gene therapy in clinical trials as well as preclinical data and include proposals on methods to further enhance the safety and efficacy of oncolytic viral vectors in cancer gene therapy.     

卵巢癌基因治疗的新进展

卵巢癌是恶性度最高的妇科肿瘤之一，由于手术和化疗的不尽人意，基因治疗是当今肿瘤研究的热点。卵巢癌的基因治疗载体主要分为病毒载体系统和非病毒载体系统。其中，腺病毒载体转染效率较高，是目前基因治疗的主要手段和最有前途的载体。卵巢癌的基因治疗策略有分子化学治疗，基因替代和免疫系统的基因调控。由于这一领域的迅速发展，卵巢癌的基因治疗有着光明的前景。     

New strategies for cancer gene therapy: Progress and opportunities

1. To date, cancer persists as one of the most devastating diseases worldwide. Problems such as metastasis and tumour resistance to chemotherapy and radiotherapy have seriously limited the therapeutic effects of existing clinical treatments.
2. To address these problems, cancer gene therapy has been developing over the past two decades, specifically designed to deliver therapeutic genes to treat cancers using vector systems. So far, a number of genes and delivery vehicles have been evaluated and significant progress has been made with several gene therapy modalities in clinical trials. However, the lack of an ideal gene delivery system remains a major obstacle for the successful translation of regimen to the clinic.
3. Recent understanding of hypoxic and necrotic regions within solid tumours and rapid development of recombinant DNA technology have reignited the idea of using anaerobic bacteria as novel gene delivery systems. These bacterial vectors have unique advantages over other delivery systems and are likely to become the vector of choice for cancer gene therapy in the near future.
4. Meanwhile, complicated tumour pathophysiology and associated metastasis make it hard to rely on a single therapeutic modality for complete tumour eradication. Therefore, the combination of cancer gene therapy with other conventional treatments has become paramount.
5. The present review introduces important cancer gene therapy strategies and major vector systems that have been studied so far with an emphasis on bacteria-mediated cancer gene therapy. In addition, exemplary combined therapies are briefly reviewed.     

肿瘤基因治疗的研究进展

癌症是基因治疗的主要对象,本文介绍了肿瘤基因治疗的方法,在基因治疗中存在的关键问题,近十几年来基因治疗的临床试验进展,并对基因治疗的安全性进行了阐述,同时分析了基因治疗的前景及未来的发展趋势。     

重组腺相关病毒载体在肺癌治疗中的应用

重组腺相关病毒（recombination Adeno-associated virus,rAAV）突出的特点使其成为基因治疗载体中的佼佼者,本文综述了rAAV基因药物在肺癌治疗中取得的进展并对其未来发展趋势进行了探讨。     

靶向基因转移系统的研究进展

基因治疗已在治疗多种人类重大疾病如遗传病、肿瘤等方面显示出良好的应用前景,但也面临着巨大的挑战,主要是如何选择安全、高效、靶向的载体系统。基因治疗的靶向性研究近年来取得了许多新的进展,如应用重组病毒载体,借助抗体或配体将治疗基因定向导入靶细胞。纳米生物材料由于其良好的生物安全性,可方便有效地实现基因的靶向性及高效表达,成为制备高效、靶向的基因治疗载体系统的良好介质,在基因治疗载体系统中日益受到广泛重视。本文综述了目前国内外在靶向基因转移系统中的最新研究进展。     

重组腺病毒介导的前列腺癌基因治疗研究进展

前列腺癌的基因治疗已逐渐成为研究热点。载体系统是基因治疗的基础。目前,腺病毒载体在前列腺癌基因治疗中应用最为广泛。溶瘤腺病毒的研究进展为前列腺癌基因治疗开辟了新的途径。近年研究表明,溶瘤腺病毒单独使用或与治疗基因联合应用,能够有效发挥抗肿瘤效应。治疗基因是发挥功能的关键因素。自杀基因能将药物转化为毒性代谢产物,细胞因子能增强抗肿瘤免疫效应,血管生成抑制因子能抑制血管生成,凋亡相关分子能诱导细胞凋亡,这些基因在前列腺癌的基础和临床研究中均表现出一定的肿瘤生长抑制作用。本文对重组腺病毒介导的前列腺癌基因治疗基础及临床研究进展进行综述。     

Adenosine deaminase deficiency as the first target disorder in gene therapy

In the past decade, the advent of gene therapy has been acclaimed as a revolutionary medical intervention, embraced with great enthusiasm. However, recent disappointing results of the considerable clinical trials have also clearly demonstrated that such an initial expectation was an overestimation of gene therapy. There are only a few successful cases despite the 3000 patients who have been treated with various forms of gene therapy. Gene therapy for severe combined immunodeficiency (SCID) caused by adenosine deaminase (ADA) deficiency is one of the few such cases where results have been promising. In particular, peripheral T-lymphocytes-directed gene therapy provides further immunological improvements for patients with ADA-SCID receiving the PEG-ADA treatment whereas gene therapy targeting haematopoietic stem cell has so far proved insufficient for clinical benefits. This report will review crucial problems elucidated in the past five clinical trials for ADA-SCID and gives an outline of the next generation of stem cell gene therapy in Japan.     

用作基因载体的阳离子脂质体及其相关材料与制备技术

综述近年来阳离子脂质体在基因转染中的应用及其相关材料和制备技术。基因疗法面临的技术问题之一是寻找合适的基因载体,阳离子脂质体作为一种非病毒载体,具有无免疫原性、可自然降解等优点,在肺部、眼部疾病以及癌症的治疗中极具应用价值。随着研究的不断深入,新型脂质体材料相继出现、制备工艺也不断更新,阳离子脂质体的理化性质和转染效率均得到了极大的改善,推动了基因疗法和基因转染研究的发展。     

Aerosol delivery of programmed cell death protein 4 using polysorbitol-based gene delivery system for lung cancer therapy

Abstract

The development of a safe and effective gene delivery system is the most challenging obstacle to the broad application of gene therapy in the clinic. In this study, we report the development of a polysorbitol-based gene delivery system as an alternative gene carrier for lung cancer therapy. The copolymer was prepared by a Michael addition reaction between sorbitol diacrylate (SD) and spermine (SPE); the SD–SPE copolymer effectively condenses with DNA on the nanoscale and protects it from nucleases. SD–SPE/DNA complexes showed excellent transfection with low toxicity both in vitro and in vivo, and aerosol delivery of SD-SPE complexes with programmed cell death protein 4 DNA significantly suppressed lung tumorigenesis in K-ras^LA1 lung cancer model mice. These results demonstrate that SD–SPE has great potential as a gene delivery system based on its excellent biocompatibility and high gene delivery efficiency for lung cancer gene therapy.     

重组复制型溶瘤腺病毒p53的质量控制方法

建立重组复制型溶瘤腺病毒p53(SG600-P53)的质控检测方法与质量标准。采用限制性内切酶酶切、PCR法对端粒酶启动子、低压缺氧调控元件融合的巨细胞病毒(cytomegalovirus,CMV)启动子、p53基因等重组病毒载体结构进行分析,鉴定结果均与理论值相符。经紫外吸收法(A260)检测,病毒颗粒数为2.0×1011 VP.mL-1;TCID50法测定感染活性为5.0×1010 IU.mL-1。p53蛋白ELISA检测结果表明,重组病毒体外感染人肺癌细胞H1299后,感染组核蛋白和空白对照组A450吸收度之比为5.2。该基因治疗制剂对人肺癌细胞A549体外杀伤的MOIIC50为1.0。以相同MOI感染并经TCID50法检测,重组病毒在人肺癌细胞A549与人表皮成纤维二倍体细胞BJ的增殖比值为398。经阴离子高效液相色谱分析,病毒载体颗粒纯度为99.5%。定量PCR检测表明在1×107 VP的病毒颗粒中,野生型腺病毒基因片段少于1个拷贝。本研究建立的重组复制型溶瘤腺病毒的质量标准与检测方法,可用于该制品的质量控制,同时也为其他溶瘤基因治疗病毒载体质控研究提供参考。     

Recent advances in aerosol gene delivery systems using non-viral vectors for lung cancer therapy

ABSTRACT

Introduction: Lung cancer commonly occurs at a high incidence worldwide. Application of aerosol gene delivery systems using various kinds of vectors can improve the patient’s quality of life by prolonging the survival rate.

Areas covered: This review provides a recent update on aerosol gene delivery strategies using various kinds of vectors and gene-modification technologies. Peptide-mediated gene therapy achieves specific targeting of cells and highly improves efficacy. Promoter-operating expression and the CRISPR/Cas9 system are novel gene therapy strategies for effective lung cancer treatment. Furthermore, hybrid systems with a combination of vectors or drugs have been recently applied as new trends in gene therapy.

Expert opinion: Although aerosol gene delivery has many advantages, physiological barriers in the lungs pose formidable challenges. Targeted gene delivery and gene-editing technology are promising strategies for lung cancer therapy. These strategies may allow the development of safety and high efficiency for clinical application. Recently, hybrid gene therapy combining novel and specific vectors has been developed as an advanced strategy. Although gene therapy for lung cancer is being actively researched, aerosol gene therapy strategies are currently lacking, and further studies on aerosol gene therapy are needed to treat lung cancer.     

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.     

年龄相关性黄斑变性基因治疗的药理与临床研究进展

年龄相关性黄斑变性（AMD）是由多因素共同诱发的视网膜黄斑区改变,基因治疗可以显著改善AMD患者临床症状,减轻长期玻璃体内治疗相关的治疗负担,提高临床适应性。尽管已取得多项成果,但基因疗法仍处于起步阶段,多数药物的临床试验仍处于研究阶段。通过整理近10年以来AMD的相关靶点治疗产品的药理研究与临床研究进展,并根据不同靶点或新载体进行分类,结合已取得的临床试验结果,评估基因治疗AMD的可行性与安全性,以期对于AMD相关靶点、治疗技术创新与载体开发提供参考。     

Gelatin-based delivery systems for cancer gene therapy

Gelatin is a natural, biocompatible, nontoxic, edible, and inexpensive macromolecule. These properties result in its wide application in pharmaceutical, medical, and cosmetic products. Recently, it has been used for the delivery of such gene therapeutic entities as plasmid DNA. This review discusses the in vivo and in vitro studies using gelatin for delivery of therapeutic genes to cancerous cells. Recent studies show that present cancer gene therapy using gelatin is lacking in both efficiency and specificity in comparison with viral vectors, whereas complexes of therapeutic DNA with modified gelatin possibly offer a safe and efficient strategy for systemic administration of therapeutic genes to solid tumors compared to injection of naked plasmid DNA. The future of these promising approaches lies in the development of better techniques for preparing gelatin–gene complexes with the aim of a gelatin-based cancer gene therapy with comparable efficiency to viral vectors but with the added advantage of biosafety for patients.     

整合素靶向性病毒载体对肿瘤细胞基因转导的促进作用

在进行基因导入研究及临床试验中，目前采用的载体通常包括了病毒及非病毒两大类。对于非病毒载体用于基因转染有许多报道。而腺病毒作为基因载体，因其具有的高效转导能力及对分裂或非分裂细胞均能转导等优点而被广泛应用于针对疾病的基因治疗中。对腺病毒进入细胞机制进行的深入研究结果发现，腺病毒与细胞结合的第一步是与细胞表面的柯萨奇腺病毒受体（coxsackie adenovirus receptor，CAR）结合，第二步是利用其在五角体基（pentonbase）中的RGD序列和细胞表面的整合素结合，     

基因药物在临床上的最新应用与研发

应用现代医学和分子药物学手段，即基因和细胞疗法，临床治疗被传统医疗视为难以攻克的病症又进入一个相对的活跃期。本文概述了近年来临床上肿瘤和神经疾病的基因治疗的实践与进展，对基因药物临床上的安全性和基因产物表达的时效性的探索以及临床基因药物研究在研发全新的抗癌病毒载体，给药途径的优化和与干细胞技术的结合提供新思路。     

基因治疗用腺相关病毒载体的剂量效应与剂量控制(英文)

人类基因治疗需要在靶细胞适时、适量、安全的产生具有疗效的药物,这些正好是定量药理学的研究范畴。重组腺相关病毒载体(rAAV)具有的各种特性以及近来在规模化制备方面取得的进展使其逐渐成为基因治疗的理想载体。超过60项以AAV为载体的基因治疗已经完成或正在展开。其中,rAAV载体的剂量与基因表达效率以及临床疗效之间的关系是研究者关注的焦点之一。多数研究认为,在一定的剂量范围内,基因表达水平具有剂量依赖性。然而,基因表达水平同时还受病毒载体构型、组织趋向性、细胞靶向性、注射途径,甚至患者的年龄、性别等条件影响,基因表达水平还可受到其他药物的调控。因此,基因治疗实验过程中应结合定量药理学深入展开,解决基因治疗中的定量与药效之间的关系。     

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。