基于高内吞效率的核酸适体构建LYTACs嵌合体以增强白血病细胞膜蛋白的降解

High-internalization-efficiency Aptamer-based LYTACs Chimera for Enhanced Degradation of Leukemia Cell Membrane Proteins

摘要

基于溶酶体靶向嵌合体(LYTACs)技术，以具有高内吞效率的转铁蛋白受体(Transferrin receptor 1，TfR1/CD71)特异性核酸适体HG9作为识别模块，通过共价连接溶酶体导向配体三乙酰半乳糖胺(tri-GalNAc)，设计合成了一种新型双功能嵌合分子HG9-10-tri，旨在实现CD71的高效、特异性降解.研究结果表明，通过筛选获得HG9与tri-GalNA之间最佳连接子长度，且该嵌合分子在不改变其稳定性及对人髓性单核细胞白血病细胞(MV4-11)亲和力的前提下，能够特异性识别急性髓细胞性白血病(AML)细胞系，并通过去唾液酸糖蛋白受体(ASGPR)介导的内吞-溶酶体途径显著下调细胞表面CD71蛋白表达水平，降解作用呈现明显的时间和浓度依赖性.并且CD71降解能够有效抑制MV4-11细胞的活力，并进一步诱导细胞凋亡与G1期周期阻滞，为治疗CD71高表达的急性髓系白血病提供了新策略.

Abstract

In this study,we developed a novel bifunctional lysosome-targeting chimera (LYTAC) ,designated HG9-10-tri,by covalently conjugating tri-GalNAc (a lysosome-trafficking ligand) to HG9,an aptamer that binds transferrin receptor 1 (TfR1/CD71) with high affinity and internalization efficiency.This chimeric molecule was designed to achieve potent and selective degradation of CD71.Following the optimization of the linker length between HG9 and tri-GalNAc,we demonstrated that HG9-10-tri retains unaltered stability and binding affinity for human acute monocytic leukemia (MV4-11) cells while specifically recognizing acute myeloid leukemia (AML) cell lines.Mechanistically,HG9-10-tri engages the asialoglycoprotein receptor (ASGPR) to trigger endocytosis and lysosomal trafficking,resulting in significant downregulation of cell surface CD71 protein levels in a time-and concentration-dependent manner.Functionally,CD71 degradation suppressed MV4-11 cell viability,induced apoptosis,and triggered G1 phase cell cycle arrest.Collectively,these findings establish HG9-10-tri as a promising therapeutic strategy for CD71-overexpressing acute myeloid leukemia.

关键词

转铁蛋白受体（CD71） / 细胞膜降解 / 溶酶体靶向嵌合体(LYTACs) / 核酸适体 / 三乙酰半乳糖胺

Key words

CD71 / Cell membrane degradation / Lysosome-targeting chimera (LYTACs) / Aptamer / Triacetylgalactosamine (tri-GalNAc)

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基金资助

国家自然科学基金(81960036)

国家自然科学基金(22564008)

贵州省教育厅高校科技创新团队([2023]068)

贵州省卫生健康委科学技术基金项目(gzwkj2021-157)

贵州省卫生健康委科学技术基金项目(2024GZWJKJXM0466)