Targeting BCL10 by small peptides for the treatment of B cell lymphoma.
Targeting BCL10 by small peptides for the treatment of B cell lymphoma.
复制标题
小肽靶向BCL10治疗B细胞淋巴瘤
作者：
Bao W;Sun C;Sun X;He M;Yu H;Yan W;Wen F;Zhang L;Yang C
原理：NF-κB信号通路的组成性激活在活化的B细胞样弥漫性大B细胞淋巴瘤（ABC-DLBCLs）的发病机制中起关键作用，ABC-DLBCLs是DLBCL最具侵袭性和耐药的形式。在ABC-DLBCL中，CARMA1-BCL10 （CB）复合物形成丝状结构，并作为构成性NF-κB活化所需的超分子组织中心（CB- smoc），使其成为ABC-DLBCL治疗的一个有吸引力的药物靶点。然而，针对CB-SMOC的药物方法一直缺乏。在这里，我们开发了专门针对Bcl10成丝过程的Bcl10肽抑制剂（BPIs）。方法：采用电镜和免疫荧光成像技术观察bpi对BCL10成丝过程的影响。通过细胞增殖试验评价所测bpi在DLBCL细胞株中的细胞毒性。通过不同的体外实验（药代动力学、免疫沉淀、免疫印迹、膜联蛋白V和PI染色）来确定bpi的作用机制。在不同的异种移植DLBCL小鼠模型中检测了bpi的体内治疗效果。最后通过Ki67、TUNEL染色和组织病理学分析来评价bpi的抗肿瘤机制和全身毒性。结果：我们发现这些bpi可以有效地破坏ABC-DLBCL细胞的BCL10成丝过程，破坏BCL10的稳定性，抑制NF-κB信号传导。通过检查一组DLBCL细胞系，我们发现这些bpi通过诱导凋亡和细胞周期阻滞来选择性地抑制cb - smoc依赖性DLBCL细胞的生长。此外，通过将BPI转化为D-retro inverso （DRI）结构，我们开发的DRI-BPI具有显著提高的细胞内稳定性和未受损的BPI活性。这些dri - bpi选择性地抑制小鼠异种移植模型中cb - smoc依赖性DLBCL肿瘤的生长，而不会引起明显的不良反应。结论：我们开发了靶向BCL10成丝过程的新型bpi，并证明bpi靶向BCL10是治疗ABC-DLBCL和其他cb - smoc依赖性恶性肿瘤的潜在安全有效的药物方法。
Rationale: Constitutive activation of the NF-κB signalling pathway plays a pivotal role in the pathogenesis of activated B cell-like diffuse large B-cell lymphomas (ABC-DLBCLs), the most aggressive and chemoresistant form of DLBCL. In ABC-DLBCLs, the CARMA1-BCL10 (CB) complex forms a filamentous structure and functions as a supramolecular organizing centre (CB-SMOC) that is required for constitutive NF-κB activation, making it an attractive drug target for ABC-DLBCL treatment. However, a pharmaceutical approach targeting CB-SMOC has been lacking. Here, we developed Bcl10 peptide inhibitors (BPIs) that specifically target the BCL10 filamentation process. Methods: Electron microscopy and immunofluorescence imaging were used to visualize the effect of the BPIs on the BCL10 filamentation process. The cytotoxicity of the tested BPIs was evaluated in DLBCL cell lines according to cell proliferation assays. Different in vitro experiments (pharmacokinetics, immunoprecipitation, western blotting, annexin V and PI staining) were conducted to determine the functional mechanisms of the BPIs. The in vivo therapeutic effect of the BPIs was examined in different xenograft DLBCL mouse models. Finally, Ki67 and TUNEL staining and histopathology analysis were used to evaluate the antineoplastic mechanisms and systemic toxicity of the BPIs. Results: We showed that these BPIs can effectively disrupt the BCL10 filamentation process, destabilize BCL10 and suppress NF-κB signalling in ABC-DLBCL cells. By examining a panel of DLBCL cell lines, we found that these BPIs selectively repressed the growth of CB-SMOC-dependent DLBCL cells by inducing apoptosis and cell cycle arrest. Moreover, by converting the BPIs to acquire a D-retro inverso (DRI) configuration, we developed DRI-BPIs with significantly improved intracellular stability and unimpaired BPI activity. These DRI-BPIs selectively repressed the growth of CB-SMOC-dependent DLBCL tumors in mouse xenograft models without eliciting discernible adverse effects. Conclusion: We developed novel BPIs to target the BCL10 filamentation process and demonstrated that targeting BCL10 by BPIs is a potentially safe and effective pharmaceutical approach for the treatment of ABC-DLBCL and other CB-SMOC-dependent malignancies.