Lipkowitz M S, Hanss B, Tulchin N, Wilson P D, Langer J C, Ross M D, Kurtzman G J, Klotman P E, Klotman M E
Department of Medicine, Mount Sinai School of Medicine, New York, New York 10029, USA. mlipkow@smtplink:mssm.edu
J Am Soc Nephrol. 1999 Sep;10(9):1908-15. doi: 10.1681/ASN.V1091908.
There has been an increasing interest recently in the possibility of treating renal diseases using gene therapy. The ability to pursue gene therapy for renal diseases has been limited by the availability of an adequate system for gene delivery to the kidney. Adeno-associated virus (AAV) is a defective virus of the parvovirus family that has a number of properties attractive for renal gene delivery: recombinant AAV contains no viral genes; expression of genes delivered by these vectors does not activate cell-mediated immunity; the virus is able to transduce nondividing as well as dividing cells; and both wild-type and recombinant AAV integrate into the host chromosome resulting in long-term gene expression. Studies were performed to determine whether AAV can deliver reporter genes to kidney cells in vitro and in vivo. These studies show that AAV can deliver reporter genes with approximately equal efficiency to human mesangial, proximal tubule, thick ascending limb, collecting tubule, and renal cell carcinoma cells in primary culture. Immortalized mouse mesangial cells are transduced at a much greater efficiency. Transduction can be enhanced by pharmaceutical agents up to sevenfold in primary cells (transducing up to 20% of primary cells per well) and as much as 400-fold in immortalized mesangial cells. AAV delivered in vivo by intraparenchymal injection results in at least 3 mo of reporter gene expression in tubular epithelial, but not glomerular or vascular, cells at the injection site. These data indicate that AAV can deliver genes to renal cells both in vitro and in vivo resulting in prolonged gene expression, and thus AAV can be a useful tool for renal gene delivery.
最近，人们对使用基因疗法治疗肾脏疾病的可能性越来越感兴趣。用于肾脏疾病的基因疗法的发展受到缺乏合适的肾脏基因递送系统的限制。腺相关病毒（AAV）是细小病毒科的一种缺陷病毒，具有许多对肾脏基因递送有吸引力的特性：重组AAV不包含病毒基因；这些载体递送的基因表达不会激活细胞介导的免疫；该病毒能够转导非分裂细胞和分裂细胞；野生型和重组AAV都能整合到宿主染色体中，从而实现长期基因表达。进行了多项研究以确定AAV是否能在体外和体内将报告基因递送至肾细胞。这些研究表明，AAV能以大致相同的效率将报告基因递送至原代培养的人系膜细胞、近端小管细胞、髓袢升支粗段细胞、集合管细胞和肾癌细胞。永生化小鼠系膜细胞的转导效率要高得多。在原代细胞中，药物可将转导效率提高多达7倍（每孔转导多达20%的原代细胞），在永生化系膜细胞中可提高多达400倍。通过实质内注射在体内递送的AAV在注射部位的肾小管上皮细胞（而非肾小球或血管细胞）中导致报告基因表达至少持续3个月。这些数据表明，AAV能在体外和体内将基因递送至肾细胞，从而实现基因的长期表达，因此AAV可成为肾脏基因递送的有用工具。
