The endosomal escape vehicle platform enhances delivery of oligonucleotides in preclinical models of neuromuscular disorders

Xiang Li; Mahboubeh Kheirabadi; Patrick G Dougherty; Kimberli J Kamer; Xiulong Shen; Nelsa L Estrella; Suresh Peddigari; Anushree Pathak; Sara L Blake; Emmanuelle Sizensky; Carmen Del Genio; Arti B Gaur; Mohanraj Dhanabal; Mahasweta Girgenrath; Natarajan Sethuraman; Ziqing Qian

doi:10.1016/j.omtn.2023.06.022

The endosomal escape vehicle platform enhances delivery of oligonucleotides in preclinical models of neuromuscular disorders

Mol Ther Nucleic Acids. 2023 Jun 29:33:273-285. doi: 10.1016/j.omtn.2023.06.022. eCollection 2023 Sep 12.

Authors

Xiang Li¹, Mahboubeh Kheirabadi¹, Patrick G Dougherty¹, Kimberli J Kamer¹, Xiulong Shen¹, Nelsa L Estrella¹, Suresh Peddigari¹, Anushree Pathak¹, Sara L Blake¹, Emmanuelle Sizensky¹, Carmen Del Genio², Arti B Gaur², Mohanraj Dhanabal¹, Mahasweta Girgenrath¹, Natarajan Sethuraman¹, Ziqing Qian¹

Affiliations

¹ Entrada Therapeutics, One Design Center Place, Suite 17-500, Boston, MA 02210, USA.
² Dartmouth Hitchcock Medical Center, 1 Medical Center Drive, Lebanon, NH 03756, USA.

Abstract

Biological therapeutic agents are highly targeted and potent but limited in their ability to reach intracellular targets. These limitations often necessitate high therapeutic doses and can be associated with less-than-optimal therapeutic activity. One promising solution for therapeutic agent delivery is use of cell-penetrating peptides. Canonical cell-penetrating peptides, however, are limited by low efficiencies of cellular uptake and endosomal escape, minimal proteolytic stability, and toxicity. To overcome these limitations, we designed a family of proprietary cyclic cell-penetrating peptides that form the core of our endosomal escape vehicle technology capable of delivering therapeutic agent-conjugated cargo intracellularly. We demonstrated the therapeutic potential of this endosomal escape vehicle platform in preclinical models of muscular dystrophy with distinct disease etiology. An endosomal escape vehicle-conjugated, splice-modulating oligonucleotide restored dystrophin protein expression in striated muscles in the mdx mouse, a model for Duchenne muscular dystrophy. Furthermore, another endosomal escape vehicle-conjugated, sterically blocking oligonucleotide led to knockdown of aberrant transcript expression levels in facioscapulohumeral muscular dystrophy patient-derived skeletal muscle cells. These findings suggest a significant therapeutic potential of our endosomal escape vehicle conjugated oligonucleotides for targeted upregulation and downregulation of gene expression in neuromuscular diseases, with possible broader application of this platform for delivery of intracellular biological agents.

Keywords: Duchenne muscular dystrophy; MT: Oligonucleotides: Therapies and Applications; antisense oligonucleotides; cell-penetrating peptides; endosomal escape; facioscapulohumeral muscular dystrophy; neuromuscular disorders.