Abstract

Saponaro, C.1, Kay, E.2, Acosta-Montalvo, A.1, Andréasson, A-C.3, Szeponik, L.4, Underwood, C.5, Stulz, R.2, Knerr, L.6, Hayen, A.6, Sengupta, K.7, Kerr-Conte, J.1, Pattou, F.1, Wallenius, K.3, Bonner C.1, Andersson, S.2 1. Univ. Lille, CHU Lille, Inserm U1190, EGID, Institut Pasteur de Lille, 59000 Lille, France 2. Discovery Sciences, R&D, AstraZeneca, Gothenburg, Sweden 3. Bioscience, Research and Early Development, Cardiovascular, Renal and Metabolism, R&D BioPharmaceuticals, AstraZeneca, Gothenburg, Sweden 4. Clinical Pathology and Safety Sciences/Imaging and Data Analytics, R&D, AstraZeneca, Gothenburg, Sweden 5. Bioscience, Research and Early Development, Cardiovascular, Renal and Metabolism, R&D BioPharmaceuticals, Cambridge, United Kingdom 6. Chemistry, Research and Early Development, Cardiovascular, Renal and Metabolism, R&D BioPharmaceuticals, AstraZeneca, Gothenburg, Sweden 7. Alliance Management, R&D, AstraZeneca, Gothenburg, Sweden Targeted delivery of oligonucleotides to specific cell types is required to achieve the broader application of oligonucleotide therapeutics. It was previously demonstrated that conjugating antisense oligonucleotide (ASO) to glucagon-like peptide 1 receptor agonist (GLP1Ra) leads to the internalization of GLP1Ra-ASO into GLP1R expressing cell lines and mouse islets, resulting in specific knockdown of the ASO target genes. However, demonstrating robust translation of GLP1Ra targeting to human pancreatic beta cells in vitro has proven difficult. This was hypothesized to be due to the poor quality of available human islets and/or insufficient GLP1R expression. For the first time, we use high-quality human islets to demonstrate human translation of GLP1R-mediated targeted delivery. We found that the islets secreted insulin in a glucose-dependent manner, which was further potentiated by the GLP1Ra-Foxo1 ASO at high-glucose concentrations. GLP1R protein expression was detected in human insulin-positive cells by immunohistochemistry (IHC). Moreover, IHC in insulin-positive cells revealed ASO accumulation after incubation with GLP1Ra-Foxo1 ASO, and qPCR revealed a significant reduction of Foxo1 mRNA expression. Overall, these data demonstrate that GLP1Ra conjugated oligonucleotides have the potential to target gene expression specifically in human beta cells.