Setegne Mekedlawit T, Cabral Aidan T, Tiwari Anushri, Shen Fangfang, Thiam Hawa Racine, Dassama Laura M K
Department of Chemistry, Stanford University, Stanford, California 94305, United States.
Sarafan ChEM-H Institute, Stanford University, Stanford, California 94305, United States.
ACS Bio Med Chem Au. 2025 Feb 27;5(2):268-282. doi: 10.1021/acsbiomedchemau.4c00098. eCollection 2025 Apr 16.
Biologics such as proteins, peptides, and oligonucleotides are powerful ligands to modulate challenging drug targets that lack readily accessible and "ligandable" pockets. However, the limited membrane permeance of biologics severely restricts their intracellular applications. Moreover, different cell types may exhibit varying levels of impermeability, and some delivery vehicles might be more sensitive to this variance. Erythroid lineage cells are especially challenging to deliver cargo to because of their unique cytoskeleton and the absence of endocytosis in mature erythrocytes. We recently employed a cell permeant miniature protein to deliver bioPROTACs to human umbilical cord blood derived erythroid progenitor cells (HUDEP-2) and primary hematopoietic stem (CD34) cells (Shen et al., ACS Cent. Sci.2022, 8, 1695-1703). While successful, the low efficiency of delivery and lack of cell-type specificity limit use of bioPROTACs . In this work, we thoroughly evaluated the performance of various recently reported cell penetrating peptides (CPPs), CPP additives, bacterial toxins, and contractile injection systems for their ability to deliver cargo to erythroid precursor cells. We also explored how targeting receptors enriched on the erythroid cell surface might improve the efficiencies and specificities of these delivery vehicles. Our results reveal that certain vehicles exhibit improved efficiencies when directed to cell surface receptors while others do not benefit from this targeting strategy. Together, these findings advance our understanding of protein delivery to challenging cell types and illustrate some of the intricacies of cell-surface receptor targeting.
蛋白质、肽和寡核苷酸等生物制品是强大的配体，可用于调节缺乏易于接近且“可配体化”口袋的具有挑战性的药物靶点。然而，生物制品有限的膜通透性严重限制了它们在细胞内的应用。此外，不同的细胞类型可能表现出不同程度的不可渗透性，并且一些递送载体可能对这种差异更敏感。由于其独特的细胞骨架以及成熟红细胞中不存在内吞作用，向红细胞谱系细胞递送货物尤其具有挑战性。我们最近使用一种细胞渗透性微型蛋白质将生物PROTAC递送至人脐带血来源的红细胞祖细胞（HUDEP-2）和原代造血干细胞（CD34）（Shen等人，《美国化学会中心科学》，2022年，8卷，1695 - 1703页）。虽然取得了成功，但递送效率低和缺乏细胞类型特异性限制了生物PROTAC的使用。在这项工作中，我们全面评估了各种最近报道的细胞穿透肽（CPP）、CPP添加剂、细菌毒素和收缩注射系统向红细胞前体细胞递送货物的能力。我们还探索了靶向富集在红细胞表面的受体如何提高这些递送载体的效率和特异性。我们的结果表明，某些载体在靶向细胞表面受体时表现出提高的效率，而其他载体则没有从这种靶向策略中受益。总之，这些发现推进了我们对向具有挑战性的细胞类型递送蛋白质的理解，并说明了细胞表面受体靶向的一些复杂性。
