Institute of Cytology, Russian Academy of Sciences, 4 Tikhoretsky Ave., St-Petersburg, 194064, Russia.
Institute of Cytology, Russian Academy of Sciences, 4 Tikhoretsky Ave., St-Petersburg, 194064, Russia.
Exp Cell Res. 2020 Apr 1;389(1):111882. doi: 10.1016/j.yexcr.2020.111882. Epub 2020 Feb 1.
The gene therapy approach aiming at curing various human diseases began to develop as a technology from early eighties of the last century. To date the delivery of therapeutic genes are mainly mediated by virus-based, predominantly, non-integrated virus vectors. These gene delivery approaches have several fundamental limitations on the way of efficient deployment in clinical gene therapy. A totally different approach was suggested about 20 years ago when episomal non-integrative artificial chromosome-based vectors featuring large size inserts (even native gene loci) advanced to the stage. Since then numerous human artificial chromosome (HAC) vectors were developed by both de novo synthesis and top-down engineering technology. This approach so far is limited to ex vivo gene transfer and correction within highly proliferative or reversibly immortalized precursor stem cells or pluripotent stem cells. Recent breakthrough in generation of induced pluripotent stem cells and embryonic stem cell manipulation give the additional pivotal stimuli to integrate it with the HAC technology and to develop thereby novel approaches to replacement therapies of human genetic diseases. The HAC technology is complex and time consuming while nowadays it has significantly advanced and become notably closer to medical applications. In this review we discuss current advancements in the HAC technology, in particular, in terms of improvement of chromosome transfer method and achievements in developing mouse-based gene therapy tissue replacement models for several monogenic human diseases. The main progress has been done in elaboration of top-down type HAC technology in modeling and preclinical studies of gene therapy treatment for Duchenne muscular dystrophy (DMD) disease.
旨在治疗各种人类疾病的基因治疗方法从上世纪 80 年代初开始作为一项技术发展起来。迄今为止，治疗基因的传递主要通过基于病毒的方法来介导，主要是非整合病毒载体。这些基因传递方法在临床基因治疗中有效应用的方式存在几个根本的局限性。大约 20 年前，当能够插入大片段（甚至是天然基因座）的基于附加体非整合人工染色体的载体作为一种全新的方法被提出时，这种情况发生了变化。从那时起，通过从头合成和自上而下的工程技术开发了许多人类人工染色体（HAC）载体。到目前为止，这种方法仅限于体外基因转移和在高度增殖或可逆永生化前体干细胞或多能干细胞内的纠正。诱导多能干细胞的最新突破和胚胎干细胞的操作给整合它与 HAC 技术并开发针对人类遗传疾病的替代治疗的新方法带来了额外的关键刺激。HAC 技术很复杂，耗时很长，但如今它已经有了显著的进步，并且明显更接近医学应用。在这篇综述中，我们讨论了 HAC 技术的最新进展，特别是在改进染色体转移方法和开发几种单基因人类疾病的基于小鼠的基因治疗组织替代模型方面的进展。在杜氏肌营养不良症（DMD）疾病的基因治疗治疗的建模和临床前研究中，主要进展是在自上而下类型的 HAC 技术的阐述方面取得的。
