靶向 NLRP3 通过诱导 PERK/eIF2 介导的细胞凋亡来抑制 AML 进展。
Targeting NLRP3 inhibits AML progression by inducing PERK/eIF2-mediated apoptosis.
发表日期:2024 Sep 02
作者:
Michela Luciano, Helene Sieberer, Peter W Krenn, Hieu-Hoa Dang, Julia Vetter, Theresa Neuper, Diana Amend, Constantin Blöchl, Christian X Weichenberger, Anna Eglseer, Michael S Unger, Ancuela Andosch, Philip Steiner, Daniel Neureiter, Renate Bauer, Laura Hummer, Suzana Tesanovic, Stephanie Binder, Dominik P Elmer, Helen Strandt, Susanne Schaller, Dirk Strunk, Lisa Pleyer, Richard Greil, Stephan Winkler, Tanja N Hartmann, Dirk Schmidt-Arras, Christian G Huber, Fritz Aberger, Jutta Horejs-Hoeck
来源:
BIOMEDICINE & PHARMACOTHERAPY
摘要:
急性髓系白血病(AML)的特点是髓系前体细胞的异常增殖,由于其异质性,给治疗带来了巨大的挑战。最近,NLRP3 炎症小体已成为 AML 发病机制的潜在贡献者,尽管其确切机制仍知之甚少。利用公共基因组数据集来评估 NLRP3 炎症小体相关基因(IL-1β、IL-18、ASC 和NLRP3)在 AML 患者中与健康个体进行比较。采用 CRISPR/Cas9 技术生成 NLRP3 缺陷的 MOLM-13 AML 细胞,然后使用实时 PCR、蛋白质印迹、FACS 分析以及透射电子和免疫荧光显微镜进行综合表征。进行蛋白质组分析以确定 NLRP3 依赖性蛋白质水平变化,重点关注 eIF2 激酶 PERK 介导的信号通路。此外,还使用白血病小鼠模型进行了体内研究,以阐明 NLRP3 在 AML 中的致病作用。NLRP3 表达升高与 AML 患者总生存期缩短显着相关。 NLRP3 的基因缺失、药理学抑制和 RNA 干扰沉默通过诱导细胞凋亡导致 AML 细胞存活率降低。蛋白质组学分析发现了 NLRP3 依赖性的蛋白质翻译改变,其特征是 NLRP3 缺陷的 AML 细胞中 eIF2α 磷酸化增强。此外,抑制 PERK 介导的 eIF2α 磷酸化可通过下调促凋亡 Bcl-2 家族成员来减少细胞凋亡。体内研究表明,移植 NLRP3 敲除 AML 细胞的小鼠的白血病负担减轻,白血病症状减轻就证明了这一点。我们的研究结果阐明了 NLRP3/PERK/eIF2 轴作为 AML 细胞存活的新驱动因素。靶向 NLRP3 诱导的信号通路,特别是通过 PERK/eIF2 轴,为 AML 干预提供了一种有前景的治疗策略。这些对 NLRP3 炎性体作用的见解为改善 AML 患者的预后和治疗结果提供了潜在途径。© 2024。作者。
Acute myeloid leukemia (AML) is characterized by the abnormal proliferation of myeloid precursor cells and presents significant challenges in treatment due to its heterogeneity. Recently, the NLRP3 inflammasome has emerged as a potential contributor to AML pathogenesis, although its precise mechanisms remain poorly understood.Public genome datasets were utilized to evaluate the expression of NLRP3 inflammasome-related genes (IL-1β, IL-18, ASC, and NLRP3) in AML patients compared to healthy individuals. CRISPR/Cas9 technology was employed to generate NLRP3-deficient MOLM-13 AML cells, followed by comprehensive characterization using real-time PCR, western blotting, FACS analysis, and transmission electron and immunofluorescence microscopy. Proteomic analyses were conducted to identify NLRP3-dependent alterations in protein levels, with a focus on the eIF2 kinase PERK-mediated signaling pathways. Additionally, in vivo studies were performed using a leukemic mouse model to elucidate the pathogenic role of NLRP3 in AML.Elevated expression of NLRP3 was significantly associated with diminished overall survival in AML patients. Genetic deletion, pharmacological inhibition and silencing by RNA interference of NLRP3 led to decreased AML cell survival through the induction of apoptosis. Proteomic analyses uncovered NLRP3-dependent alterations in protein translation, characterized by enhanced eIF2α phosphorylation in NLRP3-deficient AML cells. Moreover, inhibition of PERK-mediated eIF2α phosphorylation reduced apoptosis by downregulating pro-apoptotic Bcl-2 family members. In vivo studies demonstrated reduced leukemic burden in mice engrafted with NLRP3 knockout AML cells, as evidenced by alleviated leukemic symptoms.Our findings elucidate the involvement of the NLRP3/PERK/eIF2 axis as a novel driver of AML cell survival. Targeting NLRP3-induced signaling pathways, particularly through the PERK/eIF2 axis, presents a promising therapeutic strategy for AML intervention. These insights into the role of the NLRP3 inflammasome offer potential avenues for improving the prognosis and treatment outcomes of AML patients.© 2024. The Author(s).