Eweje Feyisayo, Ibrahim Vanessa, Shajii Aram, Walsh Michelle L, Ahmad Kiran, Alrefai Assma, Miyasato Dominie, Davis Jessie R, Ham Hyunok, Li Kaicheng, Roehrl Michael, Haller Carolyn A, Liu David R, Chen Jiaxuan, Chaikof Elliot L
Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA.
Harvard and MIT Division of Health Science and Technology, Massachusetts Institute of Technology, Cambridge, MA, USA.
Nat Biotechnol. 2025 May 15. doi: 10.1038/s41587-025-02664-2.
Intracellular delivery of biomacromolecules is hampered by low efficiency and cytotoxicity. Here we report the development of elastin-based nanoparticles for therapeutic delivery (ENTER), a recombinant elastin-like polypeptide (ELP)-based delivery system for effective cytosolic delivery of biomacromolecules in vitro and in vivo. Through iterative design, we developed fourth-generation ELPs fused to cationic endosomal escape peptides (EEPs) that self-assemble into pH-responsive micellar nanoparticles and enable cytosolic entry of cargo following endocytic uptake. In silico screening of α-helical peptide libraries led to the discovery of an EEP (EEP13) with 48% improved protein delivery efficiency versus a benchmark peptide. Our lead ELP-EEP13 showed similar or superior performance compared to lipid-based transfection reagents in the delivery of mRNA-encoded, DNA-encoded and protein-form Cre recombinase and CRISPR gene editors as well as short interfering RNAs to multiple cell lines and primary cell types. Intranasal administration of ELP-EEP13 combined with Cre protein achieved efficient editing of lung epithelial cells in reporter mice.
生物大分子的细胞内递送受到低效率和细胞毒性的阻碍。在此，我们报告了用于治疗递送的基于弹性蛋白的纳米颗粒（ENTER）的开发，这是一种基于重组弹性蛋白样多肽（ELP）的递送系统，可在体外和体内将生物大分子有效递送至细胞质。通过迭代设计，我们开发了与阳离子内体逃逸肽（EEP）融合的第四代ELP，它们自组装成pH响应性胶束纳米颗粒，并在胞吞摄取后使货物进入细胞质。对α-螺旋肽文库进行计算机筛选，发现了一种EEP（EEP13），与基准肽相比，其蛋白质递送效率提高了48%。在将mRNA编码、DNA编码和蛋白质形式的Cre重组酶以及CRISPR基因编辑器以及短干扰RNA递送至多种细胞系和原代细胞类型方面，我们的主要ELP-EEP13与基于脂质的转染试剂相比表现出相似或更优的性能。将ELP-EEP13与Cre蛋白联合进行鼻内给药，在报告基因小鼠中实现了肺上皮细胞的高效编辑。
Zotero 插件