Abstract: SA-PO0254
Antisense Oligonucleotide Distribution in Mice with Healthy and Injured Kidneys: A Single-Cell Perspective
Session Information
- Pharmacology
November 08, 2025 | Location: Exhibit Hall, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Pharmacology (PharmacoKinetics, -Dynamics, -Genomics)
- 2000 Pharmacology (PharmacoKinetics, -Dynamics, -Genomics)
Authors
- De Filippo, Elisabetta, Evotec International GmbH, Göttingen, NDS, Germany
- Bohnenpoll, Tobias, Evotec International GmbH, Göttingen, NDS, Germany
- Nordlohne, Johannes, Evotec International GmbH, Göttingen, NDS, Germany
- Cochet, Estelle, Evotec France SAS, Toulouse, Occitanie, France
- Huynh Cong, Evelyne, Evotec International GmbH, Göttingen, NDS, Germany
- Poondi Krishnan, Varsha, Evotec International GmbH, Göttingen, NDS, Germany
- Severo Witte, Maiara, Evotec International GmbH, Göttingen, NDS, Germany
- Mercuri, Annalisa, Evotec SE, Verona, Veneto, Italy
- Michi, Marco, Evotec SE, Verona, Veneto, Italy
- Seip, Britta, Evotec International GmbH, Göttingen, NDS, Germany
- Heise, Katharina Bettina, Evotec International GmbH, Göttingen, NDS, Germany
- Komarov, Ilya, Evotec International GmbH, Göttingen, NDS, Germany
- Martinez Hernandez, Ana, Evotec International GmbH, Göttingen, NDS, Germany
- Schmidt, Antje, Evotec International GmbH, Göttingen, NDS, Germany
- Wunderlich, Winfried, Evotec International GmbH, Göttingen, NDS, Germany
- Brooks, Hilary Jane, Evotec France SAS, Toulouse, Occitanie, France
- Boucher, Jeremie, Evotec International GmbH, Göttingen, NDS, Germany
- Radresa, Olivier, Evotec International GmbH, Göttingen, NDS, Germany
- Andag, Uwe, Evotec International GmbH, Göttingen, NDS, Germany
Background
Antisense oligonucleotides (ASO) are emerging as a therapeutic modality for kidney diseases. Understanding ASO distribution and silencing efficacy in different tissues and cell types in vivo is crucial for drug development. Our study aimed at characterizing and comparing the silencing efficacy and distribution at tissue and cellular resolution of two unconjugated LNA-Gapmer ASOs designed against two different genes.
Methods
Pharmacokinetics/pharmacodynamics studies were conducted using a Malat1 ASO in healthy mice and in a unilateral ischemia reperfusion injury (IRI) model. We quantified ASO accumulation and target engagement in the kidney. Single-nuclei RNA sequencing was performed to assess knockdown (KD) efficiency across the kidney cell types.
Results
In healthy mice, Malat1 ASOs accumulated in the kidney at twice the liver levels, yet tissue KD remained lower (60-80% KD in kidney and >90% KD in liver). We performed single nuclei RNA sequencing in healthy and injured kidneys and observed strong Malat1 KD in multiple kidney cell types, including proximal tubular cells, fibroblasts, intercalated cells, and glomerular cells. Furthermore, we assessed the distribution and efficacy of another ASO targeting a protein-coding RNA involved in kidney diseases. Despite the protein-coding ASO achieving only a 40% KD at organ-level (qPCR), single-nuclei RNA sequencing indicated >70% KD efficacy in several kidney cell types, including tubular cells.
Conclusion
Both ASOs induced the highest level of target KD in tubular cells in healthy and injured kidneys. Since assessing KD levels within an entire tissue provides only a partial perspective, a single-cell analysis of KD efficiency is essential to fully assess target engagement within the various kidney cell types.