美国国立卫生研究院Andre Larochelle研究组发现，移植后G-CSF通过加剧p53介导的DNA损伤反应妨碍基因编辑人类造血干细胞的植入。相关论文于2024年11月12日在线发表在《细胞—干细胞》杂志上。

研究人员发现从移植后第1天到第14天给予粒细胞集落刺激因子（G-CSF）会妨碍CRISPR-Cas9基因编辑的人类造血干细胞和祖细胞（HSPC）在小鼠异种移植模型中的植入。

G-CSF通过细胞内机制影响基因编辑的HSPC，导致增殖压力，并放大由Cas9介导的DNA双链断裂引发的早期p53介导的DNA损伤反应。

这表明存在一个阈值机制，p53的激活必须达到一个临界水平，才能损害细胞功能。暂时抑制p53或将G-CSF治疗推迟到移植后第5天，可以减轻其对基因编辑HSPC的负面影响。在CRISPR-Cas9自体HSPC基因治疗的临床试验中，考虑到移植后G-CSF可能增加HSPC的毒性，值得进一步关注。

据介绍，G-CSF常用于加速化疗后以及自体HSPC移植治疗恶性疾病后的中性粒细胞恢复。然而，其在外源性基因治疗后对人类HSPC的效果尚未被探索。

附：英文原文

Title: Post-transplant G-CSF impedes engraftment of gene-edited human hematopoietic stem cells by exacerbating p53-mediated DNA damage response

Author: Daisuke Araki, Vicky Chen, Neelam Redekar, Christi Salisbury-Ruf, Yan Luo, Poching Liu, Yuesheng Li, Richard H. Smith, Pradeep Dagur, Christian Combs, Andre Larochelle

Issue&Volume: 2024-11-12

Abstract: Granulocyte-colony-stimulating factor (G-CSF) is commonly used to accelerate recovery from neutropenia following chemotherapy and autologous transplantation of hematopoietic stem and progenitor cells (HSPCs) for malignant disorders. However, its utility after ex vivo gene therapy in human HSPCs remains unexplored. We show that administering G-CSF from day 1 to 14 post-transplant impedes engraftment of CRISPR-Cas9 gene-edited human HSPCs in murine xenograft models. G-CSF affects gene-edited HSPCs through a cell-intrinsic mechanism, causing proliferative stress and amplifying the early p53-mediated DNA damage response triggered by Cas9-mediated DNA double-strand breaks. This underscores a threshold mechanism where p53 activation must reach a critical level to impair cellular function. Transiently inhibiting p53 or delaying the initiation of G-CSF treatment to day 5 post-transplant attenuates its negative impact on gene-edited HSPCs. The potential for increased HSPC toxicity associated with post-transplant G-CSF administration in CRISPR-Cas9 autologous HSPC gene therapy warrants consideration in clinical trials.

DOI: 10.1016/j.stem.2024.10.013

Source: https://www.cell.com/cell-stem-cell/abstract/S1934-5909(24)00375-8