CD44v6 CAR-T细胞靶向dnmt3a突变AML:地西他滨的协同增强

IF 1.5 4区 医学 Q3 MEDICINE, RESEARCH & EXPERIMENTAL
Hui-Min Li, Yi-Mei Que, Xiao-Ya Cai, Ping-Fan Lu, Li-Man Lin, Min Xiao, Li Zhu, Deng-Ju Li
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引用次数: 0

摘要

目的:急性髓系白血病(Acute myeloid leukemia, AML)是一种高度异质性的疾病,DNMT3A基因突变等分子事件与AML患者预后不良相关。因此,迫切需要一种新的AML治疗方法。方法:通过RT-qPCR和Western blotting检测DNMT3A突变型AML细胞中DNMT3A mRNA和蛋白的表达。CCK-8染色和Annexin V/PI染色检测细胞增殖和凋亡。流式细胞术检测表面抗原及CD44v6 car - t细胞转染效率。构建靶向cd44v6的CAR质粒,包装慢病毒。甲基化特异性PCR用于评估启动子甲基化的差异,而ELISA用于测量细胞因子的分泌。结果:在本研究中,我们发现dnmt3a突变组细胞表面CD44v6的表达明显增加。CD44启动子区的甲基化在突变组中低于对照组。CD44v6 CAR-T细胞对dnmt3a突变的AML细胞表现出特异性的细胞毒性。低浓度地西他滨预处理可显著增强CD44v6 CAR-T细胞对dnmt3a突变AML细胞的杀伤作用(P < 0.05)。此外,地西他滨处理上调了dnmt3a突变AML细胞表面CD44v6的表达(P < 0.05)。结论:CD44v6是治疗DNMT3A突变AML患者的一种有前景的car - t细胞治疗靶点。值得注意的是,地西他滨治疗导致dnmt3a突变AML细胞表面CD44v6表达增加。这种CD44v6表达的增加促进了CD44v6 CAR-T细胞的识别和靶向。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
CD44v6 CAR-T Cells Target DNMT3A-Mutant AML: Synergistic Enhancement by Decitabine.

Objective: Acute myeloid leukemia (AML) is a highly heterogeneous disease, and molecular events such as DNMT3A gene mutations are associated with poor prognosis in AML patients. Consequently, there is an urgent need for a novel therapeutic approach for AML.

Methods: DNMT3A mRNA and protein expression were confirmed in DNMT3A-mutant AML cells via RT-qPCR and Western blotting. Cell proliferation and apoptosis were assessed via CCK-8 and Annexin V/PI staining, respectively. Flow cytometry was used to analyze surface antigens and CD44v6 CAR-T-cell transfection efficiency. CD44v6-directed CAR plasmids were constructed, and lentiviruses were packaged. Methylation-specific PCR was used to evaluate differences in promoter methylation, whereas ELISA was used to measure cytokine secretion.

Results: In this study, we found that the DNMT3A-mutant group presented significantly increased expression of CD44v6 on the cell surface. Methylation of the CD44 promoter region was lower in the mutant group than in the control group. CD44v6 CAR-T cells exhibited specific cytotoxicity against DNMT3A-mutant AML cells. Furthermore, pretreatment with low concentrations of decitabine significantly enhanced the killing effect of CD44v6 CAR-T cells on DNMT3A-mutant AML cells (P < 0.05). Additionally, decitabine treatment upregulated the expression of CD44v6 on the surface of DNMT3A-mutant AML cells (P < 0.05).

Conclusion: CD44v6 is a promising CAR-T-cell therapy target in AML patients with DNMT3A mutations. Notably, treatment with decitabine resulted in increased CD44v6 expression on the cell surface of DNMT3A-mutant AML cells. This increase in CD44v6 expression facilitates improved recognition and targeting by CD44v6 CAR-T cells.

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来源期刊
Current Medical Science
Current Medical Science Biochemistry, Genetics and Molecular Biology-Genetics
CiteScore
4.70
自引率
0.00%
发文量
126
期刊介绍: Current Medical Science provides a forum for peer-reviewed papers in the medical sciences, to promote academic exchange between Chinese researchers and doctors and their foreign counterparts. The journal covers the subjects of biomedicine such as physiology, biochemistry, molecular biology, pharmacology, pathology and pathophysiology, etc., and clinical research, such as surgery, internal medicine, obstetrics and gynecology, pediatrics and otorhinolaryngology etc. The articles appearing in Current Medical Science are mainly in English, with a very small number of its papers in German, to pay tribute to its German founder. This journal is the only medical periodical in Western languages sponsored by an educational institution located in the central part of China.
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