Jin Peipei, Chen Zhaozheng, Zhang Ju, Li Haowen, Wei Pengfei, Wang Ziyu, Feng Qiyu, Wang Hongyang, Han Da, Miao Yanyan
Department of Clinical Laboratory, Cancer Research Center, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, 230036, China; Core Unit of National Clinical Research Center for Laboratory Medicine, Hefei, Anhui, 230036, China.
Department of Clinical Laboratory, Cancer Research Center, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, 230036, China.
Acta Biomater. 2025 Mar 15;195:509-521. doi: 10.1016/j.actbio.2025.02.023. Epub 2025 Feb 14.
Various targeted protein degradation (TPD) approaches have been developed to overcome the limitations of traditional drug in eliminating pathogenic proteins by exploiting either the proteasomal or lysosomal pathway. However, there is still a lack of design strategies for TPD that utilize two distinct pathways to achieve the degradation of membrane and cytoplasmic proteins. Here, we develop a Nano-Targeting Chimera (Nano-APTAC), which is engineered by covalently attaching the protein-targeting aptamer to graphene oxide (GO) via the amide linkage, to hijack the autophagy-lysosome and ubiquitin-proteasome systems for targeted degradation of membrane and cytoplasmic proteins respectively. In contrast, a mixture of GO and aptamers without covalent interaction has no effect on protein degradation. Furthermore, the in vivo experiments demonstrate the efficacy of Nano-APTACs in depleting targeted proteins and inhibiting tumor growth. The work provides a versatile programmability platform, employing two distinct degradation systems to facilitate personalized design for the degradation of proteins regardless of their localization on the membrane or cytoplasm, and offering potential therapeutic benefits. STATEMENT OF SIGNIFICANCE: GO and aptamers have been combined for various applications. However, the utilization of this combination in TPD remains unknown. In this study, we found that the Nano-APTAC platform, constructed by covalently linking GO-aptamer chimera (not a simple mixture), can utilize autophagy-lysosome system and ubiquitin-proteasome system to degrade membrane and cytoplasmic proteins, respectively. The types of aptamers significantly influence the intracellular behavior of the chimeras, resulting in distinct subcellular localization and guiding the chimera to select specific degradation systems for protein removal. The Nano-APTAC's mode of action extremely expands the range of targeted proteins, prevents overload in specific degradation systems caused by excessive usage, and provides an exceptional level of adaptability in meeting diverse treatment requirements.
为了克服传统药物在通过蛋白酶体或溶酶体途径消除致病蛋白方面的局限性，人们开发了各种靶向蛋白质降解（TPD）方法。然而，目前仍缺乏利用两种不同途径实现膜蛋白和细胞质蛋白降解的TPD设计策略。在此，我们开发了一种纳米靶向嵌合体（Nano-APTAC），它通过酰胺键将蛋白质靶向适配体共价连接到氧化石墨烯（GO）上进行工程改造，以分别劫持自噬-溶酶体和泛素-蛋白酶体系统来靶向降解膜蛋白和细胞质蛋白。相比之下，没有共价相互作用的GO和适配体混合物对蛋白质降解没有影响。此外，体内实验证明了Nano-APTAC在消耗靶向蛋白和抑制肿瘤生长方面的有效性。这项工作提供了一个通用的可编程平台，采用两种不同的降解系统，便于针对膜或细胞质上的蛋白质降解进行个性化设计，并具有潜在的治疗益处。重要性声明：GO和适配体已被结合用于各种应用。然而，这种组合在TPD中的应用尚不清楚。在本研究中，我们发现通过共价连接GO-适配体嵌合体（而非简单混合物）构建的Nano-APTAC平台可以分别利用自噬-溶酶体系统和泛素-蛋白酶体系统来降解膜蛋白和细胞质蛋白。适配体的类型显著影响嵌合体的细胞内行为，导致不同的亚细胞定位，并引导嵌合体选择特定的降解系统来去除蛋白质。Nano-APTAC的作用模式极大地扩展了靶向蛋白的范围，防止因过度使用导致特定降解系统过载，并在满足不同治疗需求方面提供了极高的适应性。
