Design of Kidney-Targeted Nanoparticles for Delivery of Gene Therapies
- Bioengineering: Modeling, Diagnosis, Therapy
November 04, 2023 | Location: Exhibit Hall, Pennsylvania Convention Center
Abstract Time: 10:00 AM - 12:00 PM
- 400 Bioengineering
- Roach, Arantxa, The City College of New York, New York, New York, United States
- 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
- 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
Therapeutic gene and drug delivery to organ sites outside of the liver remains elusive with few strategies to do so for renal diseases. In prior studies, we found that polymeric mesoscale nanoparticles (MNPs) localize to the kidneys up to 26-fold greater than any other organ, specifically to the renal proximal tubular epithelium.
Here, we investigated the potential to adapt this MNP platform to specifically deliver various RNA-based gene therapies to the renal proximal tubules. We formulated MNPs loaded with either reporter or gene-specific siRNA, mRNA, or CRISPR gRNA/Cas9 that conformed to the 300-400 nm diameter size range which demonstrates tubular localization. We then conducted in vitro studies using renal proximal tubular epithelial cells, as well as in vivo studies with healthy mice, to confirm their function and renal-specific delivery.
Our results demonstrated successful modification of polymeric MNPs such that they maintained kidney targeting while encapsulating RNA therapeutics. mCherry mRNA-loaded MNPs exhibit rapid and bright protein translation in vitro with no cytotoxicity. GFP-targeted CRISPR gRNA/Cas9 MNPs turned off expression of GFP in a stably-transfected cell line. In vivo, we found that kidney-targeted MNPs specifically target the kidneys and achieve gene-specific knockdown.
In ongoing studies, we are using this modified MNP design to deliver gene therapies that modulate inflammation and oxidative stress in salt-sensitive hypertension models of chronic kidney disease.
a) Images of mesoscale nanoparticles. b) Kidney-specific delivery of fluorescently-labeled nucleic acids. c) GFP expression in vitro (bottom) is deleted by delivery of GFP CRISPR gRNA-Cas9 d) Renal cell lines (top) express mCherry fluorescent protein upon addition of mRNA-loaded MNPs. e) MNPs reduce gene expression specifically in the kidneys. f) Knockdown persists for up to 10 days.
- Other NIH Support