Kebenko Maxim, Zhuang Ruimeng, Hoffer Konstantin, Worthmann Anna, Horn Stefan, Kriegs Malte, Vorwerk Jan, von Bubnoff Nikolas, Khandanpour Cyrus, Gebauer Niklas, Gorantla Sivahari Prasad, Fiedler Walter, Bokemeyer Carsten, Jücker Manfred
Hubertus Wald Tumorzentrum, Department of Oncology-Hematology, Bone Marrow Transplantation and Pneumology, University Cancer Center, 20246 Hamburg, Germany.
Clinic for Hematology and Oncology, University Hospital Schleswig-Holstein Campus, 23562 Lübeck, Germany.
Int J Mol Sci. 2025 Jun 14;26(12):5721. doi: 10.3390/ijms26125721.
Acute myeloid leukemia (AML) is a heterogeneous hematological malignancy associated with a poor prognosis. Activating mutations in the FLT3 gene occur in approximately 30% of AML cases, with internal tandem duplications in the juxtamembrane domain (FLT3-ITD; 75%) and mutations in the tyrosine kinase domain (FLT3-TKD; 25%). FLT3-ITD mutations are linked to poor prognosis and offer significant clinical predictive value, whereas the implications of FLT3-TKD mutations are less understood. The Hedgehog-Gli pathway is an established therapeutic target in AML, and emerging evidence suggests crosstalk between FLT3-ITD signaling and Gli expression regulation via non-canonical mechanisms. Post-translational modifications involving myristic and palmitic acids regulate various cellular processes, but their role in AML remains poorly defined. In this study, we investigated the role of fatty acid synthase (FASN), which synthesizes myristic and palmitic acids and catalyzes palmitoyl-acyltransferation, in regulating FLT3-ITD-Gli signaling. FASN knockdown using shRNA and the FASN inhibitor TVB-3166 was performed in FLT3-ITD-mutated AML cell lines (MOLM13, MV411) and Baf3-FLT3-ITD cells. The impact of FASN inhibition was assessed through Western blot and kinome profiling, while biological implications were evaluated by measuring cell viability and proliferation. FASN inhibition resulted in reduced levels of phospho-Akt (pAkt) and phospho-S6 kinase (pS6) and decreased expression of Hedgehog-Gli1, confirming non-canonical regulation of Gli by FLT3-ITD signaling. Combining TVB-3166 with the Gli inhibitor GANT61 significantly reduced the survival of MOLM13 and MV411 cells.
急性髓系白血病（AML）是一种异质性血液系统恶性肿瘤，预后较差。FLT3基因的激活突变约发生在30%的AML病例中，其中近膜结构域的内部串联重复（FLT3-ITD；75%）和酪氨酸激酶结构域的突变（FLT3-TKD；25%）。FLT3-ITD突变与预后不良相关，并具有重要的临床预测价值，而FLT3-TKD突变的影响则了解较少。Hedgehog-Gli信号通路是AML中一个既定的治疗靶点，新出现的证据表明FLT3-ITD信号与Gli表达调控之间通过非经典机制存在相互作用。涉及肉豆蔻酸和棕榈酸的翻译后修饰调节各种细胞过程，但其在AML中的作用仍不明确。在本研究中，我们研究了合成肉豆蔻酸和棕榈酸并催化棕榈酰酰基转移的脂肪酸合酶（FASN）在调节FLT3-ITD-Gli信号中的作用。在FLT3-ITD突变的AML细胞系（MOLM13、MV411）和Baf3-FLT3-ITD细胞中使用shRNA和FASN抑制剂TVB-3166进行FASN敲低。通过蛋白质印迹和激酶组分析评估FASN抑制的影响，同时通过测量细胞活力和增殖来评估生物学意义。FASN抑制导致磷酸化Akt（pAkt）和磷酸化S6激酶（pS6）水平降低，以及Hedgehog-Gli1表达减少，证实了FLT3-ITD信号对Gli的非经典调节。将TVB-3166与Gli抑制剂GANT61联合使用可显著降低MOLM13和MV411细胞的存活率。
