Yu Gang, Chen Jing-Gui, Tan Jian-Song
Department of Hematology, Guangxi Guidong People's Hospital, Wuzhou 543001, Guangxi Zhuang Autonomous Region, China,E-mail:
Department of Hematology, Guangxi Guidong People's Hospital, Wuzhou 543001, Guangxi Zhuang Autonomous Region, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2020 Oct;28(5):1480-1485. doi: 10.19746/j.cnki.issn.1009-2137.2020.05.009.
To investigate the correlation of NK cell receptor-interacting serine/threonine kinase 1（RIPK1） activity and expression with prognosis of acute myeloid leukemia (AML) patients with FLT3-ITD mutation.
A total of 132 AML patients with FLT3-ITD mutation and 136 AML patients with FLT3-WT were selected. Clinical data and the number, length and rearrangement ratio of FLT3-ITD mutations were collected. The ratio of CD4 T cells, regulatory T cells（Tregs）, CD8 T cells, B cells, natural killer cells（NK cells） and macrophage in peripheral blood（PB） were analyzed by flow cytometry. Correlation of NK cell ratio with FLT3-ITD mutation number, length and rearrangement ratio was analyzed by Pearson correlation analysis. Western blot and RT-qPCR were used to detect RIPK1 protein and mRNA levels in NK cells, respectively. Plasma RIPK activity was detected by ELISA, and Pearson corre-lation analysis was performed for the correlation of RIPK with FLT3-ITD mutation number, length and rearrangement ratio. Kaplan-Meier curve was used to analyze the survival rate of AML patients with FLT3-ITD mutation.
Compared with AML patients with FLT3-WT, the white blood cell count in patients with FLT3-ITD mutation significantly increased, PB and BM blasts significantly decreased. There was no significant change in PB CD3 T cells, Tregs, CD8 T cells, B cells, M1 and M2 type macrophage of AML patients with FLT3-WT and FLT3-ITD mutation；Compared with AML patients with FLT3-WT, CD4 T cells significantly decreased, NK cells significantly increased in AML patients wtih FLT3-ITD mutation. NK cells ratio in AML patients with FLT3-ITD mutation significantly positively correlated with the number of FLT3-ITD mutations and rearrangement bases. Plasma RIPK1 activity, RIPK1 protein and mRNA levels in NK cells of AML patients with FLT3-ITD were significantly lower than those of AML patients with FLT3-WT, and negatively correlated with the number of FLT3-ITD mutations, the length of rearranged bases (≥52 bp) and the ratio of rearranged bases. The survival rate of AML patients with FLT3-ITD mutation in low RIPK1 activity after Rydapt treatment was significantly higher than that in high RIPK1 activity.
The prognosis of AML patients with FLT3-ITD mutation closely relates with plasma RIPK1 activity and RIPK1 expression in NK cells.
探讨NK细胞受体相互作用丝氨酸/苏氨酸激酶1（RIPK1）活性及表达与FLT3-ITD突变急性髓系白血病（AML）患者预后的相关性。
选取132例FLT3-ITD突变的AML患者和136例FLT3-WT的AML患者。收集临床资料及FLT3-ITD突变的数量、长度和重排比例。采用流式细胞术分析外周血中CD4 T细胞、调节性T细胞（Tregs）、CD8 T细胞、B细胞、自然杀伤细胞（NK细胞）和巨噬细胞的比例。采用Pearson相关分析NK细胞比例与FLT3-ITD突变数量、长度和重排比例的相关性。分别采用蛋白质免疫印迹法和RT-qPCR检测NK细胞中RIPK1蛋白和mRNA水平。采用ELISA检测血浆RIPK活性，并对RIPK与FLT3-ITD突变数量、长度和重排比例的相关性进行Pearson相关分析。采用Kaplan-Meier曲线分析FLT3-ITD突变AML患者的生存率。
与FLT3-WT的AML患者相比，FLT3-ITD突变患者的白细胞计数显著升高，外周血和骨髓原始细胞显著减少。FLT3-WT和FLT3-ITD突变的AML患者外周血CD3 T细胞、Tregs、CD8 T细胞、B细胞、M1和M2型巨噬细胞无显著变化；与FLT3-WT的AML患者相比，FLT3-ITD突变的AML患者CD4 T细胞显著减少，NK细胞显著增加。FLT3-ITD突变的AML患者NK细胞比例与FLT3-ITD突变数量和重排碱基数显著正相关。FLT3-ITD的AML患者血浆RIPK1活性、NK细胞中RIPK1蛋白和mRNA水平显著低于FLT3-WT的AML患者，且与FLT3-ITD突变数量、重排碱基长度（≥52 bp）和重排碱基比例呈负相关。Rydapt治疗后RIPK1活性低的FLT3-ITD突变AML患者的生存率显著高于RIPK1活性高的患者。
FLT3-ITD突变AML患者的预后与血浆RIPK1活性及NK细胞中RIPK1表达密切相关。
