Hematopoietic stem/progenitor cell gene therapy (HSPC-GT) has shown clear neurological benefit in rare diseases, which is achieved through the engraftment of genetically modified microglia-like cells (MLCs) in the brain. Still, the engraftment dynamics and the nature of engineered MLCs, as well as their potential use in common neurogenerative diseases, have remained largely unexplored. Here, we comprehensively characterized how different routes of administration affect the biodistribution of genetically engineered MLCs and other HSPC derivatives in mice. We generated a high-resolution single-cell transcriptional map of MLCs and discovered that they could clearly be distinguished from macrophages as well as from resident microglia by the expression of a specific gene signature that is reflective of their HSPC ontogeny and irrespective of their long-term engraftment history. Lastly, using murine models of Parkinson’s disease and frontotemporal dementia, we demonstrated that MLCs can deliver therapeutically relevant levels of transgenic protein to the brain, thereby opening avenues for the clinical translation of HSPC-GT to the treatment of major neurological diseases.

造血干/祖细胞基因疗法 (HSPC-GT) 在罕见疾病中显示出明显的神经学益处，这是通过在大脑中植入转基因小胶质细胞 (MLC) 来实现的。尽管如此，工程化 MLC 的植入动力学和性质，以及它们在常见神经生成疾病中的潜在用途，在很大程度上仍未得到探索。在这里，我们全面描述了不同的给药途径如何影响基因工程 MLC 和其他 HSPC 衍生物在小鼠体内的生物分布。我们生成了 MLC 的高分辨率单细胞转录图谱，并发现它们可以通过反映其 HSPC 个体发育的特定基因特征的表达与巨噬细胞以及常驻小胶质细胞清楚地区分开来，而与它们的长链无关。术语植入历史。最后，利用帕金森病和额颞叶痴呆的小鼠模型，我们证明 MLC 可以向大脑递送治疗相关水平的转基因蛋白，从而为 HSPC-GT 临床转化治疗主要神经系统疾病开辟途径。