Smith Laura J, Ul-Hasan Taihra, Carvaines Sarah K, Van Vliet Kim, Yang Ethel, Wong Kamehameha K, Agbandje-McKenna Mavis, Chatterjee Saswati
AAV Laboratory, Department of Virology, Beckman Research Institute of City of Hope, Duarte, California, USA.
Department of Biochemistry and Molecular Biology, Center for Structural Biology, The McKnight Brain Institute University of Florida, Gainesville, Florida, USA.
Mol Ther. 2014 Sep;22(9):1625-34. doi: 10.1038/mt.2014.107. Epub 2014 Jun 13.
Adeno-associated virus (AAV) vectors are proving to be remarkably successful for in vivo gene delivery. Based upon reports of abundant AAV in the human marrow, we tested CD34(+) hematopoietic stem cells for the presence of natural AAV. Here, we report for the first time, the presence of novel AAV variants in healthy CD34(+) human peripheral blood stem cells. The majority of healthy peripheral blood stem cell donors were found to harbor AAV in their CD34(+) cells. Every AAV isolated from CD34(+) cells mapped to AAV Clade F. Gene transfer vectors derived from these novel AAVs efficiently underwent entry and postentry processing in human cord blood stem cells and supported stable gene transfer into long-term, in vivo engrafting human HSCs significantly better than other serotypes. AAVHSC-transduced human CD34(+) cells engrafted in vivo and gave rise to differentiated transgene-expressing progeny. Importantly, gene-marked CD34(+) stem cells persisted long term in xenograft recipients, indicating transduction of primitive progenitors. Notably, correlation of structure with function permitted identification of potential capsid components important for HSC transduction. Thus, AAVHSCs represent a new class of genetic vectors for the manipulation of HSC genomes.
腺相关病毒（AAV）载体在体内基因递送方面已被证明非常成功。基于人类骨髓中存在大量AAV的报道，我们检测了CD34(+)造血干细胞中天然AAV的存在情况。在此，我们首次报告在健康的CD34(+)人外周血干细胞中存在新型AAV变体。发现大多数健康外周血干细胞供体的CD34(+)细胞中携带AAV。从CD34(+)细胞中分离出的每一种AAV都映射到AAV F进化枝。源自这些新型AAV的基因转移载体在人脐带血干细胞中能有效地进行进入和进入后处理，并且比其他血清型能更好地支持向长期体内植入的人造血干细胞进行稳定的基因转移。经AAVHSC转导的人CD34(+)细胞在体内植入并产生表达转基因的分化后代。重要的是，基因标记的CD34(+)干细胞在异种移植受体中长期存在，表明原始祖细胞被转导。值得注意的是，结构与功能的相关性使得能够鉴定出对造血干细胞转导重要的潜在衣壳成分。因此，AAVHSCs代表了一类用于操纵造血干细胞基因组的新型遗传载体。
