Consiglio Antonella, Gritti Angela, Dolcetta Diego, Follenzi Antonia, Bordignon Claudio, Gage Fred H, Vescovi Angelo Luigi, Naldini Luigi
San Raffaele Telethon Institute for Gene Therapy, Milan, Italy.
Proc Natl Acad Sci U S A. 2004 Oct 12;101(41):14835-40. doi: 10.1073/pnas.0404180101. Epub 2004 Oct 4.
Stable genetic modification of adult stem cells is fundamental for both developmental studies and therapeutic purposes. Using in vivo marking studies, we showed that injection of lentiviral vectors (LVs) into the subventricular zone of the adult mouse brain enables efficient gene transfer into long-term self-renewing neural precursors and steady, robust vector expression in their neuronal progeny throughout the subventricular zone and its rostral extension, up to the olfactory bulb. By clonal and population analysis in culture, we proved that in vivo-marked neural precursors display self-renewal and multipotency, two essential characteristics of neural stem cells (NSCs). Thus, LVs efficiently target long-term repopulating adult NSCs, and the effect of the initial transduction is amplified by the continuous generation of NSC-derived, transduced progeny. LVs may thus allow novel studies on NSCs' physiology in vivo, and introduction of therapeutic genes into NSCs may allow the development of novel approaches for untreatable CNS diseases.
成体干细胞的稳定基因改造对于发育研究和治疗目的而言都至关重要。通过体内标记研究，我们发现将慢病毒载体（LVs）注射到成年小鼠脑的脑室下区能够实现高效的基因转移，进入长期自我更新的神经前体细胞，并在其神经元后代中实现稳定、强劲的载体表达，遍及整个脑室下区及其向前延伸直至嗅球的区域。通过培养中的克隆和群体分析，我们证明体内标记的神经前体细胞具有自我更新和多能性，这是神经干细胞（NSCs）的两个基本特征。因此，慢病毒载体能够有效地靶向长期再增殖的成年神经干细胞，并且初始转导的效应会因神经干细胞衍生的转导后代的持续产生而放大。慢病毒载体因而可能允许对神经干细胞在体内的生理学进行新的研究，并且将治疗性基因引入神经干细胞可能会为无法治疗的中枢神经系统疾病开发新的方法。
