Endosomal escape cell-penetrating peptides significantly enhance pharmacological effectiveness and CNS activity of systemically administered antisense oligonucleotides
Endosomal escape cell-penetrating peptides significantly enhance pharmacological effectiveness and CNS activity of systemically administered antisense oligonucleotides
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DOI：
10.1016/j.ijpharm.2021.120398
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发表时间：
2021-03-11
影响因子：
5.8
通讯作者：
Shabanpoor, Fazel
中科院分区：
文献类型：
--
作者：
Dastpeyman, Mohadeseh;Sharifi, Ramin;Shabanpoor, Fazel
反义寡核苷酸（Antisense oligonucleotides，ASO）是近年来发展起来的一类基因特异性治疗中枢神经系统相关疾病的药物。然而，穿过血脑屏障（BBB）将阿索递送至其CNS靶细胞仍然是一个主要挑战。由于ASO主要通过内体途径被摄取到脑毛细血管内皮细胞界面中，因此内体隔室中的截留是ASO有效CNS递送的主要障碍。因此，我们评价了一组带有几个内体逃逸结构域的细胞穿透肽（CPP）对FDA批准的Spinraza（Nusinersen）（一种用于治疗脊髓性肌萎缩症（SMA）的阿索）的细胞内递送、内体释放和反义活性的有效性。我们发现了一种CPP，HA 2-Apoe（131-150），当与Nusinersen结合时，它表现出有效的内体逃逸能力，并显着增加了SMA患者来源的成纤维细胞中生存运动神经元2（SMN 2）基因的全长功能mRNA的水平。用该CPP-PMO偶联物处理SMN 2转基因成年小鼠导致脑和脊髓中全长SMN 2水平显著增加。这项工作提供了原理证明，即内体逃逸结构域与CPP的整合能够实现ASO的更高胞质递送，并且更重要的是增强全身施用的ASO的BBB渗透性和CNS活性的效率。
Antisense oligonucleotides (ASOs) are an emerging class of gene-specific therapeutics for diseases associated with the central nervous system (CNS). However, ASO delivery across the blood-brain barrier (BBB) to their CNS target cells remains a major challenge. Since ASOs are mainly taken up into the brain capillary endothelial cells interface through endosomal routes, entrapment in the endosomal compartment is a major obstacle for efficient CNS delivery of ASOs. Therefore, we evaluated the effectiveness of a panel of cell-penetrating peptides (CPPs) bearing several endosomal escape domains for the intracellular delivery, endosomal release and antisense activity of FDA-approved Spinraza (Nusinersen), an ASO used to treat spinal muscular atrophy (SMA). We identified a CPP, HA2-ApoE(131-150), which, when conjugated to Nusinersen, showed efficient endosomal escape capability and significantly increased the level of full-length functional mRNA of the survival motor neuron 2 (SMN2) gene in SMA patient-derived fibroblasts. Treatment of SMN2 transgenic adult mice with this CPP-PMO conjugate resulted in a significant increase in the level of full-length SMN2 in the brain and spinal cord. This work provides proof-of-principle that integration of endosomal escape domains with CPPs enables higher cytosolic delivery of ASOs, and more importantly enhances the efficiency of BBB-permeability and CNS activity of systemically administered ASOs.