1. Academic Validation
  2. A novel transient receptor potential C3/C6 selective activator induces the cellular uptake of antisense oligonucleotides

A novel transient receptor potential C3/C6 selective activator induces the cellular uptake of antisense oligonucleotides

  • Nucleic Acids Res. 2024 May 22;52(9):4784-4798. doi: 10.1093/nar/gkae245.
Hiroto Kohashi 1 Ryu Nagata 1 Yusuke Tamenori 2 Tomorrow Amatani 3 Yoshifumi Ueda 3 Yasuo Mori 3 Yuuya Kasahara 4 Satoshi Obika 1 5 Masahito Shimojo 1
Affiliations

Affiliations

  • 1 Graduate School of Pharmaceutical Sciences, Osaka University, Osaka 565-0871, Japan.
  • 2 School of Pharmaceutical Sciences, Osaka University, Osaka 565-0871, Japan.
  • 3 Graduate School of Engineering, Kyoto University, Kyoto 615-8510, Japan.
  • 4 National Institutes of Biomedical Innovation, Health and Nutrition, Osaka 567-0085, Japan.
  • 5 Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University, Osaka 565-0871, Japan.
Abstract

Antisense oligonucleotide (ASO) therapy is a novel therapeutic approach in which ASO specifically binds target mRNA, resulting in mRNA degradation; however, cellular uptake of ASOs remains critically low, warranting improvement. Transient receptor potential canonical (TRPC) channels regulate Ca2+ influx and are activated upon stimulation by Phospholipase C-generated diacylglycerol. Herein, we report that a novel TRPC3/C6/C7 activator, L687, can induce cellular ASO uptake. L687-induced ASO uptake was enhanced in a dose- and incubation-time-dependent manner. L687 enhanced the knockdown activity of various ASOs both in vitro and in vivo. Notably, suppression of TRPC3/C6 by specific siRNAs reduced ASO uptake in A549 cells. Application of BAPTA-AM, a Ca2+ chelator, and SKF96365, a TRPC3/C6 inhibitor, suppressed Ca2+ influx via TRPC3/C6, resulting in reduced ASO uptake, thereby suggesting that Ca2+ influx via TRPC3/C6 is critical for L687-mediated increased ASO uptake. L687 also induced dextran uptake, indicating that L687 increased endocytosis. Adding ASO to L687 resulted in endosome accumulation; however, the endosomal membrane disruptor UNC7938 facilitated endosomal escape and enhanced knockdown activity. We discovered a new function for TRPC activators regarding ASO trafficking in target cells. Our findings provide an opportunity to formulate an innovative drug delivery system for the therapeutic development of ASO.

Figures
Products
  • Cat. No.
    Product Name
    Description
    Target
    Research Area
  • HY-161766
    TRPC3/C6/C7 Activator