Diverging impact of cell fate determinants Scrib and Llgl1 on adhesion and migration of hematopoietic stem cells
Diverging impact of cell fate determinants Scrib and Llgl1 on adhesion and migration of hematopoietic stem cells
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DOI：
10.1007/s00432-018-2724-3
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发表时间：
2018-08
影响因子：
3.6
通讯作者：
Banaja P. Dash;T. Schnöder;Carolin Kathner;Juliane Mohr;Sönke Weinert;Carolin Herzog;P. S. Godavarthy;C. Zanetti;F. Perner;R. Braun-Dullaeus;B. Hartleben;T. Huber;G. Walz;M. Naumann;S. Ellis;Valera Vasioukhin;T. Kähne;D. Krause;F. Heidel
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文献类型：
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作者：
Banaja P. Dash;T. Schnöder;Carolin Kathner;Juliane Mohr;Sönke Weinert;Carolin Herzog;P. S. Godavarthy;C. Zanetti;F. Perner;R. Braun-Dullaeus;B. Hartleben;T. Huber;G. Walz;M. Naumann;S. Ellis;Valera Vasioukhin;T. Kähne;D. Krause;F. Heidel
目的细胞命运决定基因Scrib和Llgl1影响造血干细胞的自我更新能力。在连续移植和应激过程中，Scrib缺陷的造血干细胞功能受损，缺乏足够的再生能力。相反，Llgl1的缺失会导致HSC适合性增强，自我更新能力增强，干细胞库扩大。在这里，我们试图通过分析Llgl1和Scrib在造血细胞中的相互作用组来评估它们共同和独特的分子功能。方法通过亲和纯化和质谱法进行相互作用组分析。对Scrib或Llgl1条件缺失后FACS分选分离的原代小鼠HSCs的运动、迁移和黏附能力进行了检测。通过活体双光子显微镜对Scrib缺陷的HSCs进行成像。结果Scrib和Llgl1相互作用组分析显示参与了共同和独特的细胞功能。迁移和黏附是与Scrib有关的细胞功能之一，但与Llgl1无关。这些发现的功能验证证实了Scrib缺陷的HSCs在体外和体内的细胞黏附和迁移的变化。相反，Llgl1的基因失活并不影响造血干细胞的黏附和迁移能力。结论我们的数据首次提供了细胞命运决定基因Scrib在体内和体外HSC黏附和迁移中进化保守的作用的证据，这是其假定的复杂伴侣Llgl1所没有的独特功能。
PurposeCell fate determinants Scrib and Llgl1 influence self-renewal capacity of hematopoietic stem cells (HSCs). Scrib-deficient HSCs are functionally impaired and lack sufficient repopulation capacity during serial transplantation and stress. In contrast, loss of Llgl1 leads to increased HSC fitness, gain of self-renewal capacity and expansion of the stem cell pool. Here, we sought to assess for shared and unique molecular functions of Llgl1 and Scrib by analyzing their interactome in hematopoietic cells.MethodsInteractome analysis was performed by affinity purification followed by mass spectrometry. Motility, migration and adhesion were assessed on primary murine HSCs, which were isolated by FACS sorting following conditional deletion of Scrib or Llgl1, respectively. Imaging of Scrib-deficient HSCs was performed by intravital 2-photon microscopy.ResultsComparison of Scrib and Llgl1 interactome analyses revealed involvement in common and unique cellular functions. Migration and adhesion were among the cellular functions connected to Scrib but not to Llgl1. Functional validation of these findings confirmed alterations in cell adhesion and migration of Scrib-deficient HSCs in vitro and in vivo. In contrast, genetic inactivation of Llgl1 did not affect adhesion or migratory capacity of hematopoietic stem cells.ConclusionOur data provide first evidence for an evolutionarily conserved role of the cell fate determinant Scrib in HSC adhesion and migration in vitro and in vivo, a unique function that is not shared with its putative complex partner Llgl1.