背景:多重耐药(MDR)细菌引起的感染爆发,尤其是革兰氏阴性菌,已经成为医院和社区的全球性健康问题。基于反义寡核苷酸(ASO)的治疗剂对于治疗由MDR细菌引起的感染具有巨大的希望。然而,由于高分子量和亲水性导致其细胞渗透效率低,因此ASO治疗剂受到限制。
结果:这里,我们设计了一系列树枝状多肽(DPP1至DPP12)来封装ASO以形成DSPE-mPEG2000修饰的ASO/DPP纳米颗粒(DP-AD1至DP-AD12),并观察到带正电荷≥8的两亲性DP-AD2、3、7或8显示出将ASO递送到细菌中的巨大效率,但仅含两个组氨酸残基的DP-AD7和DP-AD8在体外能显著抑制细菌生长和目标基因表达。DP-AD7anti-acpP可显着提高ESBLs-E感染的脓毒症小鼠的存活率。大肠杆菌在体内表现出很强的抗菌作用。
结论:第一次,我们设计了DPP作为一种有效的载体来提供抗MDR细菌的ASO,并展示了其基本特征,即,两亲性,8-10个正电荷,和2个组氨酸残基,这是有效的基于DPP的交付所需的,为反义抗菌策略的开发和研究提供了新的途径。
BACKGROUND: Outbreaks of infection due to multidrug-resistant (MDR) bacteria, especially Gram-negative bacteria, have become a global health issue in both hospitals and communities. Antisense oligonucleotides (ASOs) based therapeutics hold a great promise for treating infections caused by MDR bacteria. However, ASOs therapeutics are strangled because of its low cell penetration efficiency caused by the high molecular weight and hydrophilicity.
RESULTS: Here, we designed a series of dendritic poly-peptides (DPP1 to DPP12) to encapsulate ASOs to form DSPE-mPEG2000 decorated ASOs/DPP nanoparticles (DP-AD1 to DP-AD12) and observed that amphipathic DP-AD2, 3, 7 or 8 with a positive charge ≥ 8 showed great efficiency to deliver ASOs into bacteria, but only the two histidine residues contained DP-AD7 and DP-AD8 significantly inhibited the bacterial growth and the targeted gene expression of tested bacteria in vitro. DP-AD7anti-acpP remarkably increased the survival rate of septic mice infected by ESBLs-E. coli, exhibiting strong antibacterial effects in vivo.
CONCLUSIONS: For the first time, we designed DPP as a potent carrier to deliver ASOs for combating MDR bacteria and demonstrated the essential features, namely, amphipathicity, 8-10 positive charges, and 2 histidine residues, that are required for efficient DPP based delivery, and provide a novel approach for the development and research of the antisense antibacterial strategy.