Deletion of Glycogen Synthase Kinase 3 Beta Reprograms NK Cell Metabolism.
Deletion of Glycogen Synthase Kinase 3 Beta Reprograms NK Cell Metabolism.
复制标题
糖原合成酶激酶3 β的缺失重新编程NK细胞代谢。
作者：
来自急性髓性白血病（AML）患者或健康供体并用IL-15扩增的自然杀伤（NK）细胞显示出缺陷的细胞毒性和升高的糖原合成酶激酶3 β（GSK 3 β）水平，并且GSK 3 β的药物抑制能够改善其细胞毒性和成熟。为了更好地了解其生物学作用，我们删除了NK细胞中的GSK 3B，并评估了它们的表型、基因表达和功能。我们没有发现细胞毒性或成熟的改变，但确实观察到代谢功能增加和线粒体代谢相关基因的改变。这表明GSK 3 β是NK细胞代谢的调节剂。细胞毒性丧失和代谢缺陷与急性髓性白血病患者或健康供体的自然杀伤（NK）细胞在IL-15体外扩增后糖原合成酶激酶3 β（GSK 3 β）过表达有关。药物抑制这些NK细胞中的GSK 3 β可提高其成熟和细胞毒活性，但GSK 3 β介导的功能障碍的机制尚未得到很好的研究。在这里，我们发现用表达膜结合IL-21的饲养细胞扩增NK细胞可以维持正常的GSK 3 β水平，这使我们能够使用CRISPR基因编辑研究GSK 3 β功能。我们删除了GSK 3 β，扩增了配对供体敲除和野生型（WT）NK细胞，然后评估了GSK 3 β缺失诱导的转录和功能改变。令人惊讶的是，我们的数据显示GSK 3B的缺失并不改变细胞毒性、细胞因子产生或成熟（如通过CD 57表达所确定的）。然而，GSK 3B-KO细胞表现出与rRNA加工、细胞增殖和代谢功能相关的基因表达的显著变化，表明可能的代谢重编程。接下来，我们发现GSK 3B-KO NK细胞中下调的关键基因在AML患者的GSK 3 β过表达NK细胞中上调，证实了临床环境中的这种相关性。最后，我们测量了细胞能量学，并观察到GSK 3B-KO NK细胞表现出150%的备用呼吸能力，这是代谢适应性的标志。这些发现表明GSK 3 β在调节NK细胞代谢中的作用。
Natural killer (NK) cells from patients with acute myeloid leukemia (AML), or from healthy donors and expanded with IL-15, show defective cytotoxicity and elevated levels of glycogen synthase kinase 3 beta (GSK3β), and drug inhibition of GSK3β was able to improve their cytotoxicity and maturation. To better understand its biologic role, we deleted GSK3B in NK cells and assessed their phenotype, gene expression, and function. We did not find alterations in cytotoxicity or maturation, but did observe increased metabolic function and alterations in genes related to mitochondrial metabolism. This suggests that GSK3β is a regulator of NK-cell metabolism. Loss of cytotoxicity and defective metabolism are linked to glycogen synthase kinase 3 beta (GSK3β) overexpression in natural killer (NK) cells from patients with acute myeloid leukemia or from healthy donors after expansion ex vivo with IL-15. Drug inhibition of GSK3β in these NK cells improves their maturation and cytotoxic activity, but the mechanisms of GSK3β-mediated dysfunction have not been well studied. Here, we show that expansion of NK cells with feeder cells expressing membrane-bound IL-21 maintained normal GSK3β levels, allowing us to study GSK3β function using CRISPR gene editing. We deleted GSK3B and expanded paired-donor knockout and wild-type (WT) NK cells and then assessed transcriptional and functional alterations induced by loss of GSK3β. Surprisingly, our data showed that deletion of GSK3B did not alter cytotoxicity, cytokine production, or maturation (as determined by CD57 expression). However, GSK3B-KO cells demonstrated significant changes in expression of genes related to rRNA processing, cell proliferation, and metabolic function, suggesting possible metabolic reprogramming. Next, we found that key genes downregulated in GSK3B-KO NK cells were upregulated in GSK3β-overexpressing NK cells from AML patients, confirming this correlation in a clinical setting. Lastly, we measured cellular energetics and observed that GSK3B-KO NK cells exhibited 150% higher spare respiratory capacity, a marker of metabolic fitness. These findings suggest a role for GSK3β in regulating NK cell metabolism.
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AML中NK细胞改变的高维质量细胞仪分析鉴定了具有不良临床结果的亚组。
DOI：
10.1073/pnas.2020459118
发表时间：
2021-06-01
影响因子：
11.1
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Chretien AS;Devillier R;Granjeaud S;Cordier C;Demerle C;Salem N;Wlosik J;Orlanducci F;Gorvel L;Fattori S;Hospital MA;Pakradouni J;Gregori E;Paul M;Rochigneux P;Pagliardini T;Morey M;Fauriat C;Dulphy N;Toubert A;Luche H;Malissen M;Blaise D;Nunès JA;Vey N;Olive D
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Olive D
影响因子：
11.2
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影响因子：
1.2
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通讯作者：
Lee, Dean A.
DOI：
10.1084/jem.20052507
发表时间：
2006-04-17
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Freud AG;Yokohama A;Becknell B;Lee MT;Mao HC;Ferketich AK;Caligiuri MA
通讯作者：
Caligiuri MA
影响因子：
7.3
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Bou-Tayeh B;Laletin V;Salem N;Just-Landi S;Fares J;Leblanc R;Balzano M;Kerdiles YM;Bidaut G;Hérault O;Olive D;Aurrand-Lions M;Walzer T;Nunès JA;Fauriat C
通讯作者：
Fauriat C