Using Nanocomplexes to Deliver Therapies for Glomerular Disease
- Genetic Diseases: Models, Mechanisms, Treatments
November 04, 2022 | Location: Exhibit Hall, Orange County Convention Center‚ West Building
Abstract Time: 10:00 AM - 12:00 PM
Category: Genetic Diseases of the Kidneys
- 1102 Genetic Diseases of the Kidneys: Non-Cystic
- Malik, Saif N., University College London, London, London, United Kingdom
- Chandler, Jennifer C., University College London, London, London, United Kingdom
- Maeshima, Ruhina, University College London, London, London, United Kingdom
- Waters, Aoife M., Great Ormond Street Hospital for Children NHS Foundation Trust, London, London, United Kingdom
- Hart, Stephen L., University College London, London, London, United Kingdom
- Kalber, Tammy L., University College London, London, London, United Kingdom
- Long, David A., University College London, London, London, United Kingdom
The onset of chronic kidney disease is often caused by damage to the kidneys’ filtration unit, the glomerulus and in particular the podocyte epithelial cells lining the outer surface of glomerular capillaries. Therefore, treatment strategies that specifically target podocytes have the potential to improve glomerular disease. For glomerular disease caused by mutations, lipid nanocomplexes provide a promising strategy as they allow nucleic acid encapsulation for gene therapy and can be conjugated with peptides for targeted delivery to receptors on podocytes such as integrin αvβ3. Therefore, we aimed to deliver lipid nanocomplexes directly to podocytes as a potential therapeutic strategy for glomerular disease.
We utilised a murine model of childhood glomerular disease with a mutation in Wilms Tumour 1 (WT1;c.1180C>T;p.R394W) to compare the ability of cyclic RGD and RWrNM peptide nanocomplexes to target integrin αvβ3 and transfect primary healthy and diseased podocytes. We first sought transcript levels of αvβ3 in podocytes compared with the rest of the kidney by qPCR before transfecting podocytes with luciferase nanocomplexes (n=3/group). Transfection was quantified by luciferase assay and normalised to total protein. We then directly compared RWrNM transfection in healthy and diseased podocytes (n=5&4). In vivo kidney targeting was assessed following a novel ultrasound guided intra-renal artery injection in healthy mice (n=3/group).
We found enrichment of αvβ3 in healthy and diseased podocytes (P=0.003 & 0.008) compared with the rest of the kidney. RWrNM nanocomplexes were more effective for αvβ3 targeting in health and disease than cyclic RGD (P=0.004 & 0.013). Furthermore, RWrNM uptake did not diminish in disease when directly compared to healthy podocytes (P=0.81). In vivo experiments show high kidney localisation following ultrasound guided delivery of luciferase cyclic RGD and RWrNM nanocomplexes with 75% and 88% of the total signal accounted for by the injected kidney respectively.
This study has demonstrated that integrin αvβ3 can be used to target nanocomplexes directly to healthy and diseased podocytes. High kidney localisation is achieved in vivo by ultrasound guided renal artery injection. This forms the basis for delivery of genes that may protect podocytes using nanocomplexes in Wt1+/R394W mice.
- Government Support – Non-U.S.