Department of Laboratory Medicine, Dongtai People's Hospital & Dongtai Hospital of Nantong University, Yancheng, P.R. China.
Department of Laboratory Medicine, Shanghai Tongji Hospital, School of Medicine, Tongji University, Shanghai, P.R. China.
Bioengineered. 2021 Dec;12(2):12383-12393. doi: 10.1080/21655979.2021.2003666.
The potential of antibodies, especially for the bispecific antibodies, are limited by high cost and complex technical process of development and manufacturing. A cost-effective and rapid platform for the endogenous antibodies expression via using the transcription (IVT) technique to produce nucleoside-modified mRNA and then encapsulated into lipid nanoparticle (LNP) may turn the body to a manufactory. Coinhibitory pathway of programmed death ligand 1 (PD-L1) and programmed cell death protein 1 receptor (PD-1) could suppress the T-cell mediated immunity. We hypothesized that the coblocking of PD-L1 and PD-1 via bispecific antibodies may achieve more potential antitumor efficacies compare with the monospecific ones. Here, we described the application of mRNA to encode a bispecific antibody with ablated Fc immune effector functions that targets both human PD-L1 and PD-1, termed XA-1, which was further assessed the functional activities and antitumor efficacies. The mRNA-encoded XA-1 held comparable abilities to fully block the PD-1/PD-L1 pathway as well as to enhance functional T cell activation compared to XA-1 protein from CHO cell source. Pharmacokinetic tests showed enhanced area under curve (AUC) of mRNA-encoded XA-1 compared with XA-1 at same dose. Chronic treatment of LNP-encapsulated XA-1 mRNA in the mouse tumor models which were reconstituted with human immune cells effectively induced promising antitumor efficacies compared to XA-1 protein. Current results collectively demonstrated that LNP-encapsulated mRNA represents the viable delivery platform for treating cancer and hold potential to be applied in the treatment of many diseases. IVT: transcription; LNP: lipid nanoparticle; hPD-1: human PD-1; hPD-L1: human PD-L1; ITS-G: Insulin-Transferrin-Selenium; Pen/Strep: penicillin-streptomycin; FBS: fetal bovine serum; TGI: tumor growth inhibition; IE1: cytomegalovirus immediate early 1; SP: signal peptide; hIgLC: human immunoglobulin kappa light chain; hIgHC: human IgG1 heavy chain; AUC: area under the curve; Cl: serum clearance; Vss: steady-state distributed volume; MLR: mixed lymphocyte reaction.
抗体，尤其是双特异性抗体，由于其开发和制造的高成本和复杂工艺，其潜力受到限制。通过使用转录（IVT）技术产生核苷修饰的 mRNA，然后将其包裹在脂质纳米颗粒（LNP）中，从而使内源性抗体表达的具有成本效益且快速的平台，可能会使身体变成一个制造厂。程序性死亡配体 1（PD-L1）和程序性死亡蛋白 1 受体（PD-1）的共抑制途径可以抑制 T 细胞介导的免疫。我们假设，通过双特异性抗体阻断 PD-L1 和 PD-1 可能比单特异性抗体具有更大的抗肿瘤疗效。在这里，我们描述了使用 mRNA 编码一种双特异性抗体的应用，该抗体的 Fc 免疫效应功能被破坏，可靶向人 PD-L1 和 PD-1，称为 XA-1，并进一步评估了其功能活性和抗肿瘤疗效。与来自 CHO 细胞源的 XA-1 蛋白相比，mRNA 编码的 XA-1 具有相同的完全阻断 PD-1/PD-L1 途径以及增强功能性 T 细胞激活的能力。药代动力学测试显示，与相同剂量的 XA-1 蛋白相比，mRNA 编码的 XA-1 的曲线下面积（AUC）增加。在用人免疫细胞重建的小鼠肿瘤模型中，慢性给予 LNP 包裹的 XA-1 mRNA 可有效诱导有希望的抗肿瘤疗效，优于 XA-1 蛋白。目前的结果共同表明，LNP 包裹的 mRNA 代表了治疗癌症的可行递送平台，并有可能应用于许多疾病的治疗。IVT：转录；LNP：脂质纳米颗粒；hPD-1：人 PD-1；hPD-L1：人 PD-L1；ITS-G：胰岛素-转铁蛋白-硒；Pen/Strep：青霉素-链霉素；FBS：胎牛血清；TGI：肿瘤生长抑制；IE1：巨细胞病毒立即早期 1；SP：信号肽；hIgLC：人免疫球蛋白κ轻链；hIgHC：人 IgG1 重链；AUC：曲线下面积；Cl：血清清除率；Vss：稳态分布体积；MLR：混合淋巴细胞反应。
