AccScience Publishing / CP / Online First / DOI: 10.36922/CP025480083
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ORIGINAL RESEARCH ARTICLE

mRNAs encoding NKG2D ligand-targeted bispecific T-cell engagers confer robust antitumor activity

Zhigang Li1 ,  Yi Wang1 ,  Qi Li1 ,  Yongzhuang Liu1 ,  Yucai Peng1*
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1 Department of Research and Development, Liverna Therapeutics Inc., Zhuhai, Guangdong, China
CP, 025480083 https://doi.org/10.36922/CP025480083
Received: 27 November 2025 | Revised: 8 January 2026 | Accepted: 22 January 2026 | Published online: 13 February 2026
© 2026 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
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Abstract

Natural killer group 2, member D (NKG2D) ligands are highly expressed in various tumor cells and therefore represent an attractive target for cancer immunotherapy. A bispecific T-cell engager (BiTE) with a fused extracellular domain of NKG2D and an anti-CD3 single-chain fragment variable (scFv) has shown significant anti-tumor potency. To circumvent the manufacturing challenges and short serum half-life of BiTEs, we developed lipid nanoparticle-encapsulated messenger RNA (mRNA) encoding α-CD3-mNKG2D fusion proteins. In vitro functional analysis revealed that the BiTE-encoded mRNA induced T cell activation and cell cytotoxicity against tumor cells. An in vivo study using an A20 lymphoma mouse model showed that BiTE-encoding mRNA significantly inhibited tumor growth. Notably, BiTE constructs without an antibody Fc domain exhibited markedly higher anti-tumor efficacy than BiTE constructs with an Fc fragment. Additionally, we designed a human version of the BiTE mRNA encoding an NKG2D and an anti-CD3 (OKT3) scFv fusion protein. Human BiTE mRNA treatment significantly upregulated CD69 expression on T cells, promoted cytotoxicity against tumor cells in vitro, and suppressed tumor growth in a peripheral blood mononuclear cell-engrafted, tumor-bearing mouse model. Based on these preclinical findings, NKG2D BiTE mRNA holds promise as a potential anti- tumor treatment meriting further clinical development.

Keywords
Messenger RNA
NKG2D
CD3
Bispecific T-cell engager
Cancer immunotherapy
Funding
None.
Conflict of interest
All authors are employed by the company Liverna Therapeutics Inc. The authors declare no conflict of interest.
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Cancer Plus, Electronic ISSN: 2661-3840 Print ISSN: 2661-3832, Published by AccScience Publishing
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