Abnormal expression of CUX1 influences autophagy activation in paroxysmal nocturnal hemoglobinuria
Abnormal expression of CUX1 influences autophagy activation in paroxysmal nocturnal hemoglobinuria
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DOI：
10.1093/jleuko/qiae008
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发表时间：
2024-02-05
影响因子：
5.5
通讯作者：
Fu，Rong
中科院分区：
文献类型：
--
作者：
Wu，Junshu;Li，Liyan;Fu，Rong
The mechanism underlying autophagy in paroxysmal nocturnal hemoglobinuria (PNH) remains largely unknown. We previously sequenced the entire genome exon of the CD59– cells from 13 patients with PNH and found genes such asCUX1encoding Cut-like homeobox 1. Peripheral blood samples from 9 patients with PNH and 7 healthy control subjects were obtained to measureCUX1expression. The correlation betweenCUX1messenger RNA expression and PNH clinical indicators was analyzed. To simulateCUX1expression in patients with PNH, we generated a panel of PNH cell lines by knocking outPIGAin K562 cell lines and transfected lentivirus withCUX1. CCK-8 and EDU assay assessed cell proliferation. Western blotting was used to detect Beclin-1, LC3A, LC3B, ULK1, PI3K, AKT, p-AKT, mTOR, and p-mTOR protein levels. Autophagosomes were observed with transmission electron microscopy. Chloroquine was used to observeCUX1expression in PNH after autophagy inhibition. Leukocytes from patients with PNH had lower levels ofCUX1messenger RNA expression and protein content than healthy control subjects. The lactose dehydrogenase level and the percentage of PNH clones were negatively correlated withCUX1relative expression. We reducedCUX1expression in aPIGAknockout K562 cell line, leading to increased cell proliferation. Levels of autophagy markers Beclin-1, LC3B, LC3A, and ULK1 increased, and autophagosomes increased. Furthermore, PI3K/AKT/mTOR protein phosphorylation levels were lower.CUX1expression did not change and cell proliferation decreased in CUX1 knocked down PNH cells after inhibition of autophagy by chloroquine. In brief,CUX1loss-of-function mutation resulted in stronger autophagy in PNH.
