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Delivery of GalNAc-Conjugated Splice-Switching ASOs to Non-hepatic Cells through Ectopic Expression of

Juergen Scharner1, Sabrina Qi1, Frank Rigo2

  • 1Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, USA.

Molecular Therapy. Nucleic Acids
|April 10, 2019
PubMed

Insights

Targeting cancer cells with splice-switching antisense oligonucleotides (ASOs) is challenging. Galactose (GalNAc)-conjugated ASOs can be delivered to cancer cells that ectopically express the asialoglycoprotein receptor (ASGP-R), enhancing therapeutic efficacy.

Area of Science:

  • Pharmacology and Therapeutics
  • Molecular Biology
  • Oncology

Background:

  • Splice-switching antisense oligonucleotides (ASOs) show therapeutic potential for genetic diseases, including cancer.
  • In vivo delivery of ASOs to tumors and specific tissues remains a significant challenge.
  • Receptor-mediated uptake, like using the asialoglycoprotein receptor (ASGP-R) with GalNAc (GN3)-conjugated ASOs in hepatocytes, offers a delivery solution.

Purpose of the Study:

  • To investigate the use of GalNAc-mediated targeting for delivering splice-switching ASOs to cancer cells.
  • To evaluate the efficacy of ASGP-R targeting in vitro and in vivo cancer models.

Main Methods:

  • Explored GalNAc-mediated targeting of splice-switching ASOs to cancer cells expressing ASGP-R ectopically.
  • Tested ASO uptake and potency in vitro across five cancer cell lines with ASGP-R1 H1a expression.
  • Evaluated in vivo targeting specificity and splicing modulation of GN3-conjugated ASOs in subcutaneous xenograft tumor models.

Main Results:

  • Ectopic expression of ASGP-R1 H1a enabled GN3-ASO uptake and increased potency in four of five tested cancer cell types.
  • Cancer cell-specific glycosylation of the receptor did not impact ASGP-R activity.
  • In vivo, GN3-conjugated ASOs specifically targeted xenograft tumors with ectopic ASGP-R1 expression, showing significantly enhanced splicing modulation compared to unconjugated ASOs.

Conclusions:

  • GalNAc-mediated targeting of ASGP-R is a viable strategy for delivering therapeutic ASOs to cancer cells with ectopic receptor expression.
  • This approach demonstrates proof-of-principle for ASO delivery in cancer models, paving the way for future strategies exploiting endogenous receptors.
Keywords:
ASGRASOASO deliveryGalNAcantisense oligonucleotideasialoglycoprotein receptorreceptor-mediated endocytosissplice switchingxenograft

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