Miller Douglas L, Pislaru Sorin V, Greenleaf James E
Department of Radiology, University of Michigan, 3315 Kresge III, 200 Zina Pitchter Place, Ann Arbor, Michigan 48109-0553, USA.
Somat Cell Mol Genet. 2002 Nov;27(1-6):115-34. doi: 10.1023/a:1022983907223.
Development of nonviral gene transfer methods would be a valuable addition to the gene-therapy armamentarium, particularly for localized targeting of specific tissue volumes. Ultrasound can produce a variety of nonthermal bioeffects via acoustic cavitation including DNA delivery. Cavitation bubbles may induce cell death or transient membrane permeabilization (sonoporation) on a single cell level, as well as microvascular hemorrhage and disruption of tissue structure. Application of sonoporation for gene delivery to cells requires control of cavitation activity. Many studies have been performed using in vitro exposure systems, for which cavitation is virtually ubiquitous. In vivo, cavitation initiation and control is more difficult, but can be enhanced by cavitation nucleation agents, such as an ultrasound contrast agent. Sonoporation and ultrasonically enhanced gene delivery has been reported for a wide range of conditions including low frequency sonication (kilohertz frequencies), lithotripter shockwaves, HIFU, and even diagnostic ultrasound (megahertz frequencies). In vitro, a variety of cell lines has been successfully transfected, with concomitant cell killing. In vivo, initial applications have been to cancer gene therapy, for which cell killing can be a useful simultaneous treatment, and to cardiovascular disease. The use of ultrasound for nonviral gene delivery has been demonstrated for a robust array of in vitro and mammalian systems, which provides a fundamental basis and strong promise for development of new gene therapy methods for clinical medicine.
非病毒基因传递方法的开发将是基因治疗手段的一项有价值的补充，特别是对于特定组织体积的局部靶向。超声可通过声空化产生多种非热生物效应，包括DNA传递。空化气泡可能在单细胞水平上诱导细胞死亡或瞬时膜通透性改变（声穿孔），以及微血管出血和组织结构破坏。将声穿孔应用于细胞基因传递需要控制空化活性。许多研究使用体外暴露系统进行，在这种系统中，空化几乎无处不在。在体内，空化的引发和控制更加困难，但可以通过空化成核剂（如超声造影剂）来增强。声穿孔和超声增强基因传递已在多种条件下得到报道，包括低频超声（千赫兹频率）、碎石冲击波、高强度聚焦超声，甚至诊断超声（兆赫兹频率）。在体外，多种细胞系已成功转染，并伴有细胞杀伤。在体内，最初的应用是癌症基因治疗，其中细胞杀伤可以作为一种有用的同步治疗方法，以及心血管疾病。超声用于非病毒基因传递已在一系列强大的体外和哺乳动物系统中得到证实，这为开发新的临床医学基因治疗方法提供了基本依据和强大前景。
