Probing the requirement for CD38 in retinoic acid-induced HL-60 cell differentiation with a small molecule dimerizer and genetic knockout.
Probing the requirement for CD38 in retinoic acid-induced HL-60 cell differentiation with a small molecule dimerizer and genetic knockout.
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DOI：
10.1038/s41598-017-17720-4
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发表时间：
2017-12-12
影响因子：
4.6
通讯作者：
Yen A
中科院分区：
文献类型：
--
作者：
MacDonald RJ;Shrimp JH;Jiang H;Zhang L;Lin H;Yen A
CD38是一种具有重要生理作用的外酶和受体。它将NAD+代谢为腺苷二磷酸核糖（ADPR）和环ADPR，调节包括钙信号在内的几个过程。CD38是白血病的阳性和阴性预后指标。在全反式维甲酸（RA）诱导的急性早幼粒细胞白血病和HL-60细胞分化中，CD38是已知最早和最显著的上调蛋白之一。CD38过表达增强分化，而morpholino和sirna诱导的敲低则减弱分化。CD38通过Src家族激酶和适配器与MAPK信号轴相互作用，促进分化。由于有证据表明CD38的重要性，我们试图确定它是否通过二聚化发挥作用。我们基于自杀底物阿拉伯糖基-2 ' -氟-2 ' -脱氧NAD+ (F-araNAD+)，二聚体F-araNAD+创建了一种连接体，以诱导同二聚化。CD38二聚化不影响ra诱导的分化。进一步探索CD38的重要性，我们创建了具有CRISPR/ cas9介导的CD38截断的HL-60细胞系。其酶结构域的缺失不影响分化。除了ra诱导的CD11b表达增加外，除CD38的前6个氨基酸外，消融所有CD38既不影响ra诱导的分化，也不影响相关的信号传导。虽然我们不能忽视这种肽的重要性，但我们的研究表明CD38对于ra诱导的分化不是必需的。
CD38 is an ectoenzyme and receptor with key physiological roles. It metabolizes NAD+ to adenosine diphosphate ribose (ADPR) and cyclic ADPR, regulating several processes including calcium signalling. CD38 is both a positive and negative prognostic indicator in leukaemia. In all-trans retinoic acid (RA)-induced differentiation of acute promyelocytic leukaemia and HL-60 cells, CD38 is one of the earliest and most prominently upregulated proteins known. CD38 overexpression enhances differentiation, while morpholino- and siRNA-induced knockdown diminishes it. CD38, via Src family kinases and adapters, interacts with a MAPK signalling axis that propels differentiation. Motivated by evidence suggesting the importance of CD38, we sought to determine whether it functions via dimerization. We created a linker based on the suicide substrate arabinosyl-2′-fluoro-2′-deoxy NAD+ (F-araNAD+), dimeric F-araNAD+, to induce homodimerization. CD38 homodimerization did not affect RA-induced differentiation. Probing the importance of CD38 further, we created HL-60 cell lines with CRISPR/Cas9-mediated CD38 truncations. Deletion of its enzymatic domain did not affect differentiation. Apart from increased RA-induced CD11b expression, ablation of all but the first six amino acids of CD38 affected neither RA-induced differentiation nor associated signalling. Although we cannot discount the importance of this peptide, our study indicates that CD38 is not necessary for RA-induced differentiation.
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