Insulin-like Growth Factor 2 (IGF2)-Fused Lysosomal Targeting Chimeras for Degradation of Extracellular and Membrane Proteins,Journal of the American Chemical Society

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Insulin-like Growth Factor 2 (IGF2)-Fused Lysosomal Targeting Chimeras for Degradation of Extracellular and Membrane Proteins
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2023-10-30 , DOI: 10.1021/jacs.3c08886
Bei Zhang ₁ , Rajeev Kungur Brahma ₂ , Liquan Zhu ₁ , Jiayi Feng ₁ , Shiqi Hu ₁ , Linghui Qian ₃ , Shubo Du ₄ , Shao Q Yao ₂ , Jingyan Ge ₁

Affiliation

Targeted degradation of the cell-surface and extracellular proteins via the endogenous lysosomal degradation pathways, such as lysosome-targeting chimeras (LYTACs), has recently emerged as an attractive tool to expand the scope of extracellular chemical biology. Herein, we report a series of recombinant proteins genetically fused to insulin-like growth factor 2 (IGF2), which we termed iLYTACs, that can be conveniently obtained in high yield by standard cloning and bacterial expression in a matter of days. We showed that both type-I iLYTACs, in which IGF2 was fused to a suitable affibody or nanobody capable of binding to a specific protein target, and type-II iLYTAC (or IGF2-Z), in which IGF2 was fused to the IgG-binding Z domain that served as a universal antibody-binding adaptor, could be used for effective lysosomal targeting and degradation of various extracellular and membrane-bound proteins-of-interest. These heterobifunctional iLYTACs are fully genetically encoded and can be produced on a large scale from conventional E. coli expression systems without any form of chemical modification. In the current study, we showed that iLYTACs successfully facilitated the cell uptake, lysosomal localization, and efficient lysosomal degradation of various disease-relevant protein targets from different mammalian cell lines, including EGFR, PD-L1, CD20, and α-synuclein. The antitumor properties of iLYTACs were further validated in a mouse xenograft model. Overall, iLYTACs represent a general and modular strategy for convenient and selective targeted protein degradation, thus expanding the potential applications of current LYTACs and related techniques.

中文翻译：

胰岛素样生长因子 2 (IGF2) 融合溶酶体靶向嵌合体用于降解细胞外和膜蛋白

通过内源性溶酶体降解途径（例如溶酶体靶向嵌合体（LYTAC））对细胞表面和细胞外蛋白质进行靶向降解，最近已成为扩大细胞外化学生物学范围的有吸引力的工具。在此，我们报道了一系列与胰岛素样生长因子 2 (IGF2) 基因融合的重组蛋白，我们将其称为 iLYTAC，可以通过标准克隆和细菌表达在几天内方便地以高产量获得。我们展示了 I 型 iLYTAC（其中 IGF2 与能够结合特定蛋白质靶点的合适亲和体或纳米抗体融合）和 II 型 iLYTAC（或 IGF2-Z）（其中 IGF2 与 IgG 融合）结合 Z 结构域作为通用抗体结合接头，可用于有效的溶酶体靶向和降解各种细胞外和膜结合的目的蛋白。这些异双功能 iLYTAC 是完全基因编码的，可以从常规大肠杆菌表达系统大规模生产，无需任何形式的化学修饰。在当前的研究中，我们表明 iLYTAC 成功促进了来自不同哺乳动物细胞系的各种疾病相关蛋白靶标的细胞摄取、溶酶体定位和有效溶酶体降解，包括 EGFR、PD-L1、CD20 和 α-突触核蛋白。iLYTAC 的抗肿瘤特性在小鼠异种移植模型中得到了进一步验证。总体而言，iLYTAC 代表了一种通用的模块化策略，可方便且选择性地靶向蛋白质降解，从而扩展了当前 LYTAC 和相关技术的潜在应用。

更新日期：2023-10-30

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