Chen Chun-Yu, Vander Kooi Amber, Cavedon Alex, Cai Xiaohe, Hoggatt Jonathan, Martini Paolo G V, Miao Carol H
Seattle Children's Research Institute, Seattle, WA 98101, USA.
Moderna, Inc., Cambridge, MA 02139, USA.
Mol Ther Nucleic Acids. 2023 Oct 5;34:102043. doi: 10.1016/j.omtn.2023.102043. eCollection 2023 Dec 12.
Development of factor VIII (FVIII) inhibitors is a serious complication in the treatment of hemophilia A (HemA) patients. In clinical trials, anti-CD3 antibody therapy effectively modulates the immune response of allograft rejection or autoimmune diseases without eliciting major adverse effects. In this study, we delivered mRNA-encapsulated lipid nanoparticles (LNPs) encoding therapeutic anti-CD3 antibody (αCD3 LNPs) to overcome the anti-FVIII immune responses in HemA mice. It was found that αCD3 LNPs encoding the single-chain antibodies (Fc-scFv) can efficiently deplete CD3 and CD4 effector T cells, whereas αCD3 LNPs encoding double-chain antibodies cannot. Concomitantly, mice treated with αCD3 (Fc-scFv) LNPs showed an increase in the CD4CD25Foxp3 regulatory T cell percentages, which modulated the anti-FVIII immune responses. All T cells returned to normal levels within 2 months. HemA mice treated with αCD3 LNPs prior to hydrodynamic injection of liver-specific FVIII plasmids achieved persistent FVIII gene expression without formation of FVIII inhibitors. Furthermore, transgene expression was increased and persistent following secondary plasmid challenge, indicating induction of long-term tolerance to FVIII. Moreover, the treated mice maintained their immune competence against other antigens. In conclusion, our study established a potential new strategy to induce long-term antigen-specific tolerance using an αCD3 LNP formulation.
因子 VIII（FVIII）抑制剂的产生是甲型血友病（HemA）患者治疗中的严重并发症。在临床试验中，抗 CD3 抗体疗法可有效调节同种异体移植排斥反应或自身免疫性疾病的免疫反应，且不会引发重大不良反应。在本研究中，我们递送了编码治疗性抗 CD3 抗体的 mRNA 封装脂质纳米颗粒（LNPs）（αCD3 LNPs），以克服 HemA 小鼠体内的抗 FVIII 免疫反应。结果发现，编码单链抗体（Fc-scFv）的αCD3 LNPs 能够有效清除 CD3 和 CD4 效应 T 细胞，而编码双链抗体的αCD3 LNPs 则不能。同时，用αCD3（Fc-scFv）LNPs 处理的小鼠 CD4CD25Foxp3 调节性 T 细胞百分比增加，从而调节了抗 FVIII 免疫反应。所有 T 细胞在 2 个月内恢复到正常水平。在经流体动力学注射肝脏特异性 FVIII 质粒之前用αCD3 LNPs 处理的 HemA 小鼠实现了持续的 FVIII 基因表达，且未形成 FVIII 抑制剂。此外，在二次质粒攻击后，转基因表达增加并持续，表明对 FVIII 诱导了长期耐受性。此外，处理后的小鼠对其他抗原保持免疫能力。总之，我们的研究建立了一种使用αCD3 LNP 制剂诱导长期抗原特异性耐受性的潜在新策略。
