Abstract: TH-PO0043
Mesoscale Lipid Nanoparticles for Targeted Delivery of Gene Therapy in CKD
Session Information
- Bioengineering: MPS, Flow, and Delivery
November 06, 2025 | Location: Exhibit Hall, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Bioengineering
- 400 Bioengineering
Authors
- Vasylaki, Anastasiia, The City College of New York, New York, New York, United States
- Ghosh, Pratyusha, The City College of New York, New York, New York, United States
- Sookraj, Shakuntala Devi, The City College of New York, New York, New York, United States
- Jaimes, Edgar A., Memorial Sloan Kettering Cancer Center, New York, New York, United States
- Williams, Ryan, The City College of New York, New York, New York, United States
Background
Gene therapies are showing significant clinical value and hold great promise for nephrology. However, delivering nucleic acids to the kidneys is challenging due to rapid degradation and physiological clearance. A potential solution involves using kidney-targeting nanoparticles. Previously, we showed that mesoscale nanoparticles (300–500 nm) selectively accumulate in the tubular epithelium. Additionally, ionizable lipid nanoparticles have been shown to overcome some barriers for nucleic acid delivery. Therefore, our goal was to develop a new lipid-based mesoscale nanoparticle for gene therapy in chronic kidney disease (CKD).
Methods
We prepared a novel mesoscale lipid nanoparticle (MLNP) formulation containing siRNA, an ionizable lipid, a phospholipid, cholesterol, and a PEG lipid via nanoprecipitation. Particle size, PDI, and ζ-potential were characterized using dynamic light scattering. siRNA quantification was performed with the RiboGreen assay. Biocompatibility was assessed using the MTT assay and siRNA release was evaluated for sustained delivery. For biodistribution studies, C57BL/6J mice received IV injections of fluorescently labeled MLNPs. Organs were collected at 1, 3, 5, and 7 days post-injection for fluorescent imaging. Gene knockdown efficacy was analyzed in the organs using RT-qPCR.
Results
Five formulation modifications were tested to increase the size of LNPs to the mesoscale range, with the most significant effect observed by increasing total lipid concentration. To ensure stable PEGylation required for kidney targeting, the lipid composition of MLNPs was optimized. Cytotoxicity assay in 786-O and HK-2 cells demonstrated minimal effect of MLNPs on cell viability. The siRNA release profile showed sustained release over 72h. The biodistribution study demonstrated a 3-fold higher accumulation in the kidneys vs the liver in 24h and up to an 85-fold difference after 7 days. Target gene mRNA levels in the kidneys decreased by about 20-30%, indicating effective gene knockdown.
Conclusion
This study presents, to our knowledge, the first mesoscale lipid nanoparticle formulation optimized for nucleic acid delivery to the kidneys. MLNPs exhibited high kidney selectivity and achieved targeted gene knockdown with minimal toxicity. Further research will focus on applying this delivery system for CKD treatment.
Funding
- Other NIH Support