Adaptation to ex vivo culture drives human haematopoietic stem cell loss of repopulation capacity in a cell cycle independent manner
Adaptation to ex vivo culture drives human haematopoietic stem cell loss of repopulation capacity in a cell cycle independent manner
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DOI：
10.1101/2022.11.17.516906
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发表时间：
2023-01
期刊：
影响因子：
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通讯作者：
Carys Johnson;Kendig Sham;Serena Belluschi;Xiaonan Wang;Winnie W. Y. Lau;K. Kaufmann;Gabriela Krivdova-Gabriela
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文献类型：
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作者：
Carys Johnson;Kendig Sham;Serena Belluschi;Xiaonan Wang;Winnie W. Y. Lau;K. Kaufmann;Gabriela Krivdova-Gabriela
长期造血干细胞功能（LT-HSC）的丧失阻碍了体外HSC基因治疗和扩增过程的成功，但发生这种情况的动力学和机制仍未完全表征。通过时间分辨scRNA-Seq，匹配体内功能分析和使用可逆的G1早期停在体外系统，我们定义了人类lt - hsc第一次离体分裂过程中发生的转录和功能事件的序列。我们证明，与目前的假设相反，在培养过程中长期再生能力的丧失与细胞周期进程无关。相反，它是一个快速的事件，伴随着对文化的早期适应，其特征是短暂的基因表达动态和基因表达的有限全球变异性。然而，细胞周期的进展有助于在培养中建立分化程序。我们的数据对改进HSC基因治疗和扩展方案具有重要意义。
Loss of long-term haematopoietic stem cell function (LT-HSC) hampers the success of ex vivo HSC gene therapy and expansion procedures, but the kinetics and the mechanisms by which this occurs remain incompletely characterized. Here through time-resolved scRNA-Seq, matched in vivo functional analysis and the use of a reversible in vitro system of early G1 arrest, we define the sequence of transcriptional and functional events occurring during the first ex vivo division of human LT-HSCs. We demonstrate that contrary to current assumptions, loss of long-term repopulation capacity during culture is independent of cell cycle progression. Instead it is a rapid event that follows an early period of adaptation to culture, characterised by transient gene expression dynamics and constrained global variability in gene expression. Cell cycle progression however contributes to the establishment of differentiation programmes in culture. Our data have important implications for improving HSC gene therapy and expansion protocols.