Milani Michela, Fabiano Anna, Perez-Rodriguez Marta, Hernandez Raisa Jofra, Zecchillo Alessandra, Zonari Erika, Ottonello Sofia, Basso-Ricci Luca, Canepari Cesare, Volpin Monica, Iannello Valeria, Capo Valentina, Quaranta Pamela, Seffin Luca, Russo Fabio, Biffi Mauro, Ormoli Leonardo, Brombin Chiara, Carlucci Filippo, Forlino Antonella, Filibian Marta, Montini Eugenio, Scala Serena, Villa Anna, Bueren Juan Antonio, Rio Paula, Aiuti Alessandro, Cantore Alessio, Naldini Luigi
San Raffaele Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific Institute, Milan, Italy.
Biomedical Innovation Unit, Center for Research on Energy, Environment and Technology (CIEMAT), Madrid, Spain.
Nature. 2025 May 28. doi: 10.1038/s41586-025-09070-3.
Lentiviral vector (LV)-mediated ex vivo gene therapy for haematopoietic stem and progenitor cells (HSPCs) has delivered on the promise of a 'one-and-done' treatment for several genetic diseases. However, ex vivo manipulation and patient conditioning before transplantation are major hurdles that could be overcome by an in vivo approach. Here we demonstrate that in vivo gene delivery to HSPCs after systemic LV administration is enabled by the substantial trafficking of these cells from the liver to the bone marrow in newborn mice. We improved gene-transfer efficiency using a phagocytosis-shielded LV, successfully reaching bona fide HSPCs capable of long-term multilineage output and engraftment after serial transplantation, as confirmed by clonal tracking. HSPC mobilization further increased gene transfer, extending the window of intervention, although permissiveness to LV transduction declined with age. We successfully tested this in vivo strategy in mouse models of adenosine deaminase deficiency, autosomal recessive osteopetrosis and Fanconi anaemia. Interestingly, in vivo gene transfer provided a selective advantage to corrected HSPCs in Fanconi anaemia, leading to near-complete haematopoietic reconstitution and prevention of bone marrow failure. Given that circulating HSPCs in humans are also most abundant shortly after birth, in vivo HSPC gene transfer holds strong translational potential across multiple diseases.
慢病毒载体（LV）介导的造血干细胞和祖细胞（HSPCs）体外基因治疗已实现了对多种遗传性疾病进行“一次治疗，终身受益”的承诺。然而，移植前的体外操作和患者预处理是主要障碍，而体内方法可以克服这些障碍。在这里，我们证明，在新生小鼠中，全身给予LV后，HSPCs会大量从肝脏迁移到骨髓，从而实现对HSPCs的体内基因递送。我们使用一种吞噬作用屏蔽的LV提高了基因转移效率，通过克隆追踪证实，成功地转导了真正的HSPCs，这些细胞在连续移植后能够实现长期多谱系输出和植入。HSPC动员进一步提高了基因转移效率，延长了干预窗口，尽管随着年龄的增长，对LV转导的易感性会下降。我们在腺苷脱氨酶缺乏症、常染色体隐性遗传性骨质石化症和范可尼贫血的小鼠模型中成功测试了这种体内策略。有趣的是，体内基因转移为范可尼贫血中校正的HSPCs提供了选择性优势，导致几乎完全的造血重建并预防了骨髓衰竭。鉴于人类循环中的HSPCs在出生后不久也最为丰富，体内HSPC基因转移在多种疾病中具有强大的转化潜力。
