PO.ET02.08 · 实验与分子治疗
LIN-T（脂质整合标签）：一种用于靶向治疗性mRNA递送的优化标签-脂质纳米颗粒设计
LIN-T (lipid integrated tag): An optimized tag-lipid nanoparticle design for targeted therapeutic mRNA delivery
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：脂质纳米颗粒（LNP）是mRNA递送的领先平台，但靶向递送仍具挑战性。脂质组成、颗粒大小、电荷和可电离脂质选择等因素会影响生物分布、细胞摄取和mRNA翻译。抗体修饰的LNP提供了一种拓展治疗应用的策略。
方法：传统的抗体修饰方法，如配制后偶联（点击化学或马来酰亚胺化学）、融合蛋白或位点特异性脂质偶联，都需要对抗体进行化学修饰。这些修饰可能损害稳定性并增加聚集风险。为解决这一问题，我们开发了一种模块化系统，将一个短肽标签整合到LNP表面，通过双特异性抗体设计实现快速抗体连接，无需化学偶联。
创新点：我们的9个氨基酸标签（在我们近期发表于Nature Communications的文章中优化：PMID 39500897）以皮摩尔亲和力结合scFv（单链可变片段）结构域。该标签在体外和体内显示出极低的免疫原性风险，可能是由于标签在结合时被屏蔽。我们展示了两种互补的标签整合策略：
1. 基于蛋白质：重组惰性ApoE（ApoEi）-融合标签，用于对预制LNP进行快速包被。2. 基于合成脂质：在LNP组装过程中整合DSPE-PEG-标签偶联物。
两种方法均可保持标签的可及性并实现高效的抗体修饰。ApoEi-标签设计独特之处还在于可阻断ApoE-LDL受体相互作用，我们能够展示这可降低肝脏趋向性。
结果：利用该平台，我们在Her2+、CD40+、CD3+和CD20+细胞中实现了靶向mRNA递送和翻译。合成DSPE-PEG-标签支持可扩展、GMP兼容的生产，而基于ApoEi的方法为研究目的提供了一种快速工具。我们的人源scFv可格式化为经典的Morrison双特异性设计，并在CHO细胞中以>8 g/L的滴度表达。
结论：这些策略为精准靶向mRNA递送提供了一个多功能框架，支持在肿瘤学和自身免疫性疾病等领域中可定制的LNP疗法。
查看英文原文 English abstract
Background: Lipid nanoparticles (LNPs) are the leading platform for mRNA delivery, but targeted delivery remains challenging. Factors such as lipid composition, particle size, charge, and ionizable lipid selection influence biodistribution, cellular uptake, and mRNA translation. Antibody-decorated LNPs offer a strategy to expand therapeutic applications.
Approach: Conventional antibody decoration methods, such as post-formulation conjugation (click or maleimide chemistry), fusion proteins, or site-specific lipid conjugation, require chemical modification of antibodies. Modifications that can compromise stability and increase aggregation risk. To address this, we developed a modular system using a short peptide tag integrated into the LNP surface, enabling rapid antibody attachment via a bispecific antibody design without chemical conjugation.
Innovation: Our 9-amino-acid tag (optimized in our recent Nature com. publication: PMID 39500897) binds an scFv (single-chain variable fragment) domain with picomolar affinity. The tag shows minimal immunogenicity risk in vitro and in vivo, likely as the tag is shielded upon binding. We demonstrate two complementary strategies for tag incorporation on the LNP:
1. Protein-based: Recombinant inert ApoE (ApoEi)-fusion tag for rapid coating of preformed LNPs.2. Synthetic lipid-based: DSPE-PEG-tag conjugate integrated during LNP assembly.
Both approaches preserve tag accessibility and enable efficient antibody decoration. The ApoEi-tag design uniquely also blocks ApoE-LDL receptor interactions, and we can display that this reduces liver tropism.
Results: Using this platform, we achieved targeted mRNA delivery and translation in Her2+, CD40+, CD3+, and CD20+ cells. The synthetic DSPE-PEG-tag supports scalable, GMP-compatible production, and the ApoEi-based method enables a rapid tool for research purposes. Our human scFv can be formatted into a classical Morrison bispecific design and expressed in CHO cells at titers >8 g/L.
Conclusion: These strategies provide a versatile framework for precision-targeted mRNA delivery, supporting customizable LNP therapeutics within for example oncology and autoimmune diseases.
利益披露 Disclosure
T. Lidström, None.
A. Kostakis,
Strike Pharma AB Employment, Other, Industrial PhD student.
R. Veerman,
Strike Pharma AB Employment, Stock Option.
V. Emruli,
Strike Pharma AB Employment, Stock Option.
S. Guleed,
Strike Pharma AB Employment, Stock Option.
T. Furebring,
Strike Pharma AB Independent Contractor, Stock.
J. A. Nilsson,
Circular biotech AB Stock, Patent.
S. Mangsbo,
Strike Pharma AB Stock, ), Other Intellectual Property.