Isolation of DNA Aptamer Targeting PD-1 with an Antitumor Immunotherapy Effect

Immune checkpoints play a vital role in regulating T cell responses. Programmed cell death 1 (PD-1), a key inhibitory immune-checkpoint receptor, negatively regulates the human immune response. Anti-PD-1 therapy is an immune-checkpoint inhibition therapy, which is a progressing clinical strategy in treating various human cancers. Aptamers, called "chemical antibodies", have several virtues, including better tissue penetration, lower immunogenicity, and ease of production. Here, after 10 rounds of selection using engineered cells with PD-1 overexpression as target cells, we successfully isolated four anti-PD-1 aptamer candidates using cell-SELEX (systematic evolution of ligands by exponential enrichment) procedure. Among them, the candidate PD4S showed the highest affinity with an equilibrium dissociation constant (K_d) of 10.3 nM and rescued the T cell function suppressed by PD-1/PD-L1. Treatment of PD4S in the CT26 carcinoma model showed an antitumor effect. Together, the anti-PD-1 aptamer PD4S could be applied as an alternative agent in immunotherapy.