Endosomal escape cell-penetrating peptides significantly enhance pharmacological effectiveness and CNS activity of systemically administered antisense oligonucleotides

Mohadeseh Dastpeyman; Ramin Sharifi; Azin Amin; John A Karas; Brittany Cuic; Yijun Pan; Joseph A Nicolazzo; Bradley J Turner; Fazel Shabanpoor

doi:10.1016/j.ijpharm.2021.120398

Endosomal escape cell-penetrating peptides significantly enhance pharmacological effectiveness and CNS activity of systemically administered antisense oligonucleotides

Int J Pharm. 2021 Apr 15:599:120398. doi: 10.1016/j.ijpharm.2021.120398. Epub 2021 Feb 26.

Authors

Mohadeseh Dastpeyman¹, Ramin Sharifi¹, Azin Amin¹, John A Karas², Brittany Cuic¹, Yijun Pan³, Joseph A Nicolazzo³, Bradley J Turner⁴, Fazel Shabanpoor⁵

Affiliations

¹ The Florey Institute of Neuroscience and Mental Health, University of Melbourne, Parkville 3010, VIC, Australia.
² School of Chemistry, The University of Melbourne, VIC 3010, Australia; The Bio21 Institute, University of Melbourne, 30 Flemington Rd., VIC 3010, Australia.
³ Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville 3052, VIC, Australia.
⁴ The Florey Institute of Neuroscience and Mental Health, University of Melbourne, Parkville 3010, VIC, Australia; The Perron Institute for Neurological and Translational Science, Queen Elizabeth Medical Centre, Nedlands, Western Australia 6150, Australia.
⁵ The Florey Institute of Neuroscience and Mental Health, University of Melbourne, Parkville 3010, VIC, Australia. Electronic address: fazel.shabanpoor@unimelb.edu.au.

PMID: 33640427
DOI: 10.1016/j.ijpharm.2021.120398

Abstract

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.

Keywords: Antisense Therapy; Antisense oligonucleotides; Blood-brain barrier; Cell-penetrating peptides; Endosomal escape.

MeSH terms

Animals
Cell-Penetrating Peptides*
Central Nervous System
Endothelial Cells
Humans
Mice
Muscular Atrophy, Spinal*
Oligonucleotides, Antisense

Substances

Cell-Penetrating Peptides
Oligonucleotides, Antisense