In vivo human T cell engineering with enveloped delivery vehicles.
In vivo human T cell engineering with enveloped delivery vehicles.
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DOI：
10.1038/s41587-023-02085-z
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发表时间：
2024-01
影响因子：
46.9
通讯作者：
J. Hamilton;Evelyn Chen;Barbara S. Perez;Cindy R Sandoval Espinoza;Min Hyung Kang;M. Trinidad;Wayne Ngo;Jennifer A. Doudna
中科院分区：
文献类型：
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作者：
J. Hamilton;Evelyn Chen;Barbara S. Perez;Cindy R Sandoval Espinoza;Min Hyung Kang;M. Trinidad;Wayne Ngo;Jennifer A. Doudna
病毒和病毒衍生的颗粒具有向细胞传递分子的内在能力，但容易改变细胞类型选择性的困难阻碍了它们用于治疗传递。在这里，我们证明了通过包裹CRISPR-Cas9蛋白和引导RNA的膜源性颗粒上显示的抗体片段识别细胞表面标记物可以将基因组编辑工具传递到特定细胞。与依赖进化的衣壳趋向性递送病毒编码货物的腺相关病毒等传统载体相比，这些cas9包装包膜递送载体（cas9 - edv）利用可预测的抗体-抗原相互作用，选择性地将基因组编辑机制短暂地递送到感兴趣的细胞。抗体靶向的cas9 - edv在体外和体内混合群体中优先赋予同源靶细胞基因组编辑功能，而不是旁观者细胞。通过使用多重靶向分子直接递送到人类T细胞，cas9 - edv能够在人源化小鼠中产生基因组编辑的嵌合抗原受体T细胞，建立了一种具有广泛治疗用途潜力的可编程递送方式。
Viruses and virally derived particles have the intrinsic capacity to deliver molecules to cells, but the difficulty of readily altering cell-type selectivity has hindered their use for therapeutic delivery. Here, we show that cell surface marker recognition by antibody fragments displayed on membrane-derived particles encapsulating CRISPR–Cas9 protein and guide RNA can deliver genome editing tools to specific cells. Compared to conventional vectors like adeno-associated virus that rely on evolved capsid tropisms to deliver virally encoded cargo, these Cas9-packaging enveloped delivery vehicles (Cas9-EDVs) leverage predictable antibody–antigen interactions to transiently deliver genome editing machinery selectively to cells of interest. Antibody-targeted Cas9-EDVs preferentially confer genome editing in cognate target cells over bystander cells in mixed populations, both ex vivo and in vivo. By using multiplexed targeting molecules to direct delivery to human T cells, Cas9-EDVs enable the generation of genome-edited chimeric antigen receptor T cells in humanized mice, establishing a programmable delivery modality with the potential for widespread therapeutic utility.