Targeted Lipid Nanoparticles in Nucleic Acid Delivery

Posted on July 02, 2024

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The development of lipid nanoparticles (LNPs) has revolutionized the field of nucleic acid delivery, offering a promising approach to treat various diseases. Targeted lipid nanoparticles (tLNPs) enhance the precision and efficacy of delivering a variety of therapeutic agents including nucleic acids to specific tissues and cell populations, thereby improving therapeutic outcomes and minimizing off-target effects. LNPs are essential for efficiently delivering and safeguarding mRNA to cells, and they are currently in the limelight as essential parts of the COVID-19 mRNA vaccines. This blog explores the advancements in tLNPs, focusing on their design, mechanisms of action, and potential applications in treating diverse medical conditions.

What are Targeted-Lipid Nanoparticles?

Lipid nanoparticles (LNPs) are advanced drug delivery systems composed of lipids like cholesterol, helper lipids, polyethylene glycol (PEG)-lipids, and ionizable lipids. These components form a stable, biocompatible vehicle that encapsulates and protects nucleic acids such as mRNA and siRNA, ensuring efficient delivery into target cells for gene expression or silencing. The 'targeted' aspect of tLNPs involves modifying these nanoparticles with ligands or antibodies to bind specifically to receptors on target cells, ensuring precise delivery. LNPs are the most efficient and advanced non-viral delivery platforms for nucleic acids.

Figure 1: Structure of Lipid Nanoparticle

The Evolution of Lipid Nanoparticles

The journey of LNPs began with the direct intramuscular injection of naked mRNA into mice, a technique that demonstrated limited efficacy due to the large, polar, and unstable nature of nucleic acids. The introduction of LNPs addressed these challenges by enhancing the stability and cellular uptake of nucleic acids. Today, LNPs are the gold standard for nucleic acid delivery, with several FDA-approved applications, including COVID-19 mRNA vaccines and treatments for hereditary amyloidosis.

The Chemistry of LNPs

LNPs are formulated through a process called alcohol dilution, where nucleic acids are dispersed in an aqueous buffer and lipids are dissolved in alcohol. This mixture is rapidly combined, typically using microfluidic mixing, to create nanoparticles that encapsulate the nucleic acids. Targeted LNPs are further refined by incorporating targeting ligands or modifying lipid compositions to enhance specificity to desired tissues or cells.

Targeting Mechanisms

LNPs can be passively or actively targeted to specific tissues.

  • Passive targeting relies on the physicochemical properties of the nanoparticles, such as size and charge. For instance, smaller and negatively charged LNPs are more effectively taken up by lymph node dendritic cells, making them suitable for vaccine delivery.

    The size of LNPs can be tuned by adjusting the molar amounts of lipids like DMG-PEG2000. Increasing the amount of DMG-PEG2000 from 0.004 µmol to 0.12 µmol reduces the LNP size from 200 nm to 30 nm, significantly enhancing cellular uptake by CD+ dendritic cells in lymph nodes.

    Passive targeting takes advantage of the enhanced permeability and retention (EPR) effect, which is particularly useful in targeting tumors. Tumor tissues typically have leaky vasculature and poor lymphatic drainage, allowing nanoparticles to accumulate more in tumor tissue than in normal tissues. Additionally, the surface charge and hydrophilicity of LNPs can be tuned to enhance their interaction with specific cell types or tissues, improving their uptake and retention in the desired locations.

  • Active or ligand-mediated targeting, on the other hand, involves the addition of specific ligands or antibodies to the LNP surface. These ligands bind to receptors on the target cells, ensuring precise delivery. For example, mannose-conjugated LNPs have been shown to enhance dendritic cell uptake, boosting immune responses in vaccine applications.

    Active targeting of LNPs using antibodies is achieved by incorporating DSPE-PEG2000-maleimide, a PEGylated lipid that acts as a linker for attaching antibodies or targeting ligands to the LNP surface. DSPE-PEG2000-maleimide enables the conjugation of antibodies like anti-CD4 and PECAM-1, facilitating targeted delivery to T cells and lung vascular endothelial cells. This approach enhances the specificity and efficiency of LNP delivery, reducing off-target effects and improving therapeutic outcomes. Leveraging DSPE-PEG2000-maleimide, researchers can design LNPs for precise delivery to various tissues, expanding applications in fields such as oncology and neurology.
Overcoming Hepatic Tropism

One of the challenges with LNPs is their natural tendency to accumulate in the liver, primarily due to scavenger receptor-mediated uptake by hepatic cells. While this is beneficial for treating liver diseases, it limits the application of LNPs for non-hepatic targets. Researchers have developed strategies to overcome this challenge, such as altering the surface charge and composition of LNPs or using "nanoprimers" to temporarily block liver uptake, thereby enhancing delivery to other tissues.

Applications in Cancer Therapy

Lipid nanoparticles hold significant promise in oncology, particularly for delivering mRNA-based cancer vaccines and immunotherapies. Companies like BioNTech and Moderna are pioneering this field with several mRNA cancer vaccines currently in clinical trials. These vaccines are designed to activate the immune system against tumor-specific antigens, offering a novel approach to cancer treatment.

Expanding Applications Beyond the Liver

The potential applications of tLNPs extend far beyond liver diseases. For instance, tLNPs are being explored for targeted delivery to the lungs, which could revolutionize the treatment of respiratory diseases such as cystic fibrosis and influenza. Similarly, targeting LNPs to the central nervous system (CNS) could provide new avenues for treating neurological disorders by overcoming the blood-brain barrier. Moreover, tLNPs are being used to deliver CRISPR-Cas9 components, offering a potential for precise gene editing in diseased cells.

Conclusion and Future Directions

Targeted lipid nanoparticles represent a cutting-edge advancement in the field of drug delivery, with the potential to transform the treatment of various diseases. Future research will likely focus on improving the specificity and efficiency of tLNPs, as well as exploring new applications across different medical fields. As the technology continues to evolve, tLNPs may soon become a cornerstone of personalized medicine, offering targeted and effective treatments tailored to individual patient needs.

The development and refinement of tLNPs are crucial for advancing medical science and improving patient outcomes. Continued research and innovation in this field hold the promise of unlocking new therapeutic possibilities and addressing unmet medical needs.

Revolutionize ligand-mediated targeting with DSPE-PEG2000-maleimide from Avanti in your lipid nanoparticles.

Click here for the full article: Kularatne RN, Crist RM, Stern ST. The Future of Tissue-Targeted Lipid Nanoparticle-Mediated Nucleic Acid Delivery. Pharmaceuticals (Basel). 2022 Jul 20;15(7):897. doi: 10.3390/ph15070897. PMID: 35890195; PMCID: PMC9322927.