Institute of Molecular and Cell Biology, Singapore, 138673, Singapore.
Department of Biochemistry, National University of Singapore, Singapore, 119077, Singapore.
Sci Rep. 2020 Oct 20;10(1):17762. doi: 10.1038/s41598-020-73998-x.
Biologics such as peptides and antibodies are a well-established class of therapeutics. However, their intracellular delivery remains problematic. In particular, methods to efficiently inhibit intra-nuclear targets are lacking. We previously described that Pseudomonas Exotoxin A reaches the nucleoplasm via the endosomes-to-nucleus trafficking pathway. Here, we show that a non-toxic truncated form of PE can be coupled to peptides and efficiently reach the nucleoplasm. It can be used as a Peptide Nuclear Delivery Device (PNDD) to deliver polypeptidic cargos as large as Glutathione- S-transferase (GST) to the nucleus. PNDD1 is a fusion of PNDD to the c-myc inhibitor peptide H1. PNDD1 is able to inhibit c-Myc dependent transcription at nanomolar concentration. In contrast, H1 fused to various cell-penetrating peptides are active only in the micromolar range. PNDD1 attenuates cell proliferation and induces cell death in various tumor cell lines. In particular, several patient-derived Diffuse Large B-Cell Lymphomas cell lines die after exposure to PNDD1, while normal B-cells survive. Altogether, our data indicate that PNDD is a powerful tool to bring active cargo to the nucleus and PNDD1 could be the basis of a new therapy against lymphoma.
生物制剂，如肽和抗体，是一类成熟的治疗药物。然而，它们的细胞内传递仍然存在问题。特别是，缺乏有效抑制核内靶标的方法。我们之前描述了绿脓杆菌外毒素 A 通过内体到核的运输途径到达核质。在这里，我们表明，一种非毒性的 PE 截断形式可以与肽结合，并有效地到达核质。它可以用作肽核传递装置 (PNDD) 将像谷胱甘肽-S-转移酶 (GST) 这样的大的多肽货物递送到细胞核。PNDD1 是 PNDD 与 c-myc 抑制剂肽 H1 的融合。PNDD1 能够以纳摩尔浓度抑制 c-Myc 依赖性转录。相比之下，与各种穿透肽融合的 H1 仅在微摩尔范围内具有活性。PNDD1 可抑制多种肿瘤细胞系的增殖并诱导细胞死亡。特别是，几种源自患者的弥漫性大 B 细胞淋巴瘤细胞系在暴露于 PNDD1 后死亡，而正常 B 细胞存活。总之，我们的数据表明，PNDD 是将活性货物带入细胞核的有力工具，PNDD1 可能是治疗淋巴瘤的新疗法的基础。
