2. Academic Validation
  3. Electrostatic adsorption of polyanions onto lipid nanoparticles controls uptake, trafficking, and transfection of RNA and DNA therapies

Electrostatic adsorption of polyanions onto lipid nanoparticles controls uptake, trafficking, and transfection of RNA and DNA therapies

  • Proc Natl Acad Sci U S A. 2024 Mar 12;121(11):e2307809121. doi: 10.1073/pnas.2307809121.
Namita Nabar 1 2 3,  Tamara G Dacoba 1 3,  Gil Covarrubias 1,  Denisse Romero-Cruz 4 5,  Paula T Hammond 1 2 3
Affiliations

Affiliations

  • 1 Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139.
  • 2 Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139.
  • 3 Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, MA 02139.
  • 4 Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139.
  • 5 Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139.
PMID: 38437543 DOI: 10.1073/pnas.2307809121
Abstract

Rapid advances in nucleic acid therapies highlight the immense therapeutic potential of genetic therapeutics. Lipid nanoparticles (LNPs) are highly potent nonviral transfection agents that can encapsulate and deliver various nucleic acid therapeutics, including but not limited to messenger RNA (mRNA), silencing RNA (siRNA), and plasmid DNA (pDNA). However, a major challenge of targeted LNP-mediated systemic delivery is the nanoparticles' nonspecific uptake by the liver and the mononuclear phagocytic system, due partly to the adsorption of endogenous serum proteins onto LNP surfaces. Tunable LNP surface chemistries may enable efficacious delivery across a range of organs and cell types. Here, we describe a method to electrostatically adsorb bioactive polyelectrolytes onto LNPs to create layered LNPs (LLNPs). LNP cores varying in nucleic acid cargo and component lipids were stably layered with four biologically relevant polyanions: hyaluronate (HA), poly-L-aspartate (PLD), poly-L-glutamate (PLE), and polyacrylate (PAA). We further investigated the impact of the four surface polyanions on the transfection and uptake of mRNA- and pDNA-loaded LNPs in cell cultures. PLD- and PLE-LLNPs increased mRNA transfection twofold over unlayered LNPs in immune cells. HA-LLNPs increased pDNA transfection rates by more than twofold in epithelial and immune cells. In a healthy C57BL/6 murine model, PLE- and HA-LLNPs increased transfection by 1.8-fold to 2.5-fold over unlayered LNPs in the liver and spleen. These results suggest that LbL assembly is a generalizable, highly tunable platform to modify the targeting specificity, stability, and transfection efficacy of LNPs, as well as incorporate other charged targeting and therapeutic molecules into these systems.

Keywords

gene delivery; layer-by-layer; lipid nanoparticles; messenger RNA.

Figures
Products