Engineered mRNA-expressed bispecific antibody prevent intestinal cancer via lipid nanoparticle delivery

ABSTRACT

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 in vitro 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 in vitro functional activities and in vivo antitumor efficacies. The in vitro 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.

Abbreviations: IVT: in vitro 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.

KEYWORDS:

• PD-1
• PD-L1
• mRNA
• LNP
• bispecific antibody
• cancer immunotherapy

Highlight

• A mRNA-encoded bispecific antibody (XA-1) targeted PD-1 and PD-L1 has been created.

• A single dose of XA-1 mRNA-LNPs exerted notably prolonged XA-1 persistence in vivo.

• XA-1 mRNA-LNPs therapy obviously inhibited the growth of colorectal carcinoma.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Authors’ contributions

Conceptualization: Shihai Xuan, Anquan Shang; Data curation: Lipei Wu, Weiwei Wang, Jiale Tian; Methodology: Lipei Wu, Weiwei Wang, Jiale Tian, Chunrun Qi, Zhengxin Cai; Resources: Shihai Xuan, Anquan Shang; Software: Zhengxin Cai, Wenhui Yan; Supervision: Shihai Xuan, Anquan Shang; Validation: Lipei Wu, Weiwei Wang, Jiale Tian, Chunrun Qi; Writing-original draft: Lipei Wu; Writing - review & editing: Weiwei Wang, Jiale Tian, Chunrun Qi, Zhengxin Cai, Wenhui Yan, Shihai Xuan, Anquan Shang.

Additional information

Funding

The author(s) reported there is no funding associated with the work featured in this article.