State Key Laboratory of Cancer Biology, Department of Biochemistry and Molecular Biology, The Fourth Military Medical University, Xi'an, Shaanxi 710032, China.
Department of Biochemistry and Molecular Biology, Zunyi Medical College, Zunyi, Guizhou 563000, China.
Sci Rep. 2016 Jun 22;6:28352. doi: 10.1038/srep28352.
Chronic myeloid leukemia (CML) is characterized by constitutively active fusion protein tyrosine kinase BCR-ABL. Although the tyrosine kinase inhibitor (TKI) against BCR-ABL, imatinib, is the first-line therapy for CML, acquired resistance almost inevitably emerges. The underlying mechanism are point mutations within the BCR-ABL gene, among which T315I is notorious because it resists to almost all currently available inhibitors. Here we took use of a previously generated chimeric ubiquitin ligase, SH2-U-box, in which SH2 from the adaptor protein Grb2 acts as a binding domain for activated BCR-ABL, while U-box from CHIP functions as an E3 ubiquitin ligase domain, so as to target the ubiquitination and degradation of both native and T315I-mutant BCR-ABL. As such, SH2-U-box significantly inhibited proliferation and induced apoptosis in CML cells harboring either the wild-type or T315I-mutant BCR-ABL (K562 or K562R), with BCR-ABL-dependent signaling pathways being repressed. Moreover, SH2-U-box worked in concert with imatinib in K562 cells. Importantly, SH2-U-box-carrying lentivirus could markedly suppress the growth of K562-xenografts in nude mice or K562R-xenografts in SCID mice, as well as that of primary CML cells. Collectively, by degrading the native and T315I-mutant BCR-ABL, the chimeric ubiquitin ligase SH2-U-box may serve as a potential therapy for both imatinib-sensitive and resistant CML.
慢性髓性白血病 (CML) 的特征是组成性激活融合蛋白酪氨酸激酶 BCR-ABL。尽管针对 BCR-ABL 的酪氨酸激酶抑制剂 (TKI) 伊马替尼是 CML 的一线治疗药物,但几乎不可避免地会出现获得性耐药。其潜在机制是 BCR-ABL 基因内的点突变,其中 T315I 最为臭名昭著,因为它几乎抵抗所有现有的抑制剂。在这里,我们利用了先前生成的嵌合泛素连接酶 SH2-U-box,其中来自衔接蛋白 Grb2 的 SH2 作为激活的 BCR-ABL 的结合域,而 CHIP 的 U-box 作为 E3 泛素连接酶结构域,从而靶向天然和 T315I 突变的 BCR-ABL 的泛素化和降解。因此,SH2-U-box 显著抑制了携带野生型或 T315I 突变 BCR-ABL(K562 或 K562R)的 CML 细胞的增殖并诱导其凋亡,同时抑制了 BCR-ABL 依赖性信号通路。此外,SH2-U-box 与伊马替尼在 K562 细胞中协同作用。重要的是,携带 SH2-U-box 的慢病毒可以显著抑制裸鼠中的 K562 异种移植物或 SCID 小鼠中的 K562R 异种移植物以及原代 CML 细胞的生长。总之,通过降解天然和 T315I 突变的 BCR-ABL,嵌合泛素连接酶 SH2-U-box 可能成为治疗伊马替尼敏感和耐药 CML 的潜在疗法。