通过抑制翻译来扩大人类造血干细胞的扩张
Chenchen Li1, Hanna Shin1, Dheeraj Bhavanasi1
1Division of Hematology-Oncology, Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
bioRxiv : the preprint server for biology
|December 11, 2023
概括
使用转化抑制剂改善了用于移植的血造干细胞 (HSC) 扩张. 这种方法支持ex vivo HSC扩张和血液疾病的基因编辑.
科学领域:
- 血液学 血液学 血液学
- 干细胞生物学 干细胞生物学
- 分子生物学分子生物学
背景情况:
- 从带血 (UCB) 移植血造干细胞 (HSC) 对于治疗血液疾病至关重要,但由于HSC数量较低而受到限制.
- 对基因编辑的ex vivo HSC操纵受到HSC损失的阻碍,因为扩张必不可少的细胞因子通过增加蛋白质合成来损害自我更新.
- 之前确定的无细胞因子方法保持HSC,但不会扩展它们.
研究的目的:
- 确定从UCB获得人体HSCs的ex vivo扩张方法,同时保持它们的自我更新能力.
- 为了克服低HSC数量和ex vivo操纵诱导的损失的局限性.
- 为了促进HSC的基因编辑,用于治疗血红蛋白病等遗传性血液疾病.
主要方法:
- 高通量查以确定与高细胞扩张相容的翻译抑制剂.
- 在低细胞因子条件下与已识别的翻译抑制剂相结合培养人类HSCs.
- 使用移植试验和单细胞转录组分析评估HSC扩张.
- 评估该方法在维持和扩展基因编辑HSC方面的有效性.
主要成果:
- 特定的转化抑制剂的鉴定,使人类HSCs能够在体外扩张.
- 在培养一周内,从UCB获得长期HSC的约5倍扩张.
- 单细胞转录组分析证实了HSC的维持,由未折叠的蛋白质应激反应介质的表达证明.
- 证明了CRISPR/Cas9编辑HSC的成功维护和扩展,包括那些针对BCL11A+58增强器的HSC.
结论:
- 开发了一种新的方法,用于使用翻译抑制剂和低细胞因子条件进行人体HSCs的ex vivo扩张.
- 这种方法成功地维持和扩展了CRISPR/Cas9基因编辑后的HSC,解决了对血红蛋白病变的基因校正的一个关键挑战.
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