Protein and mRNA Delivery Enabled by Cholesteryl‐Based Biodegradable Lipidoid Nanoparticles

Abstract

Developing safe and efficient delivery systems for therapeutic biomacromolecules is a long‐standing challenge. Herein, we report a newly developed combinatorial library of cholesteryl‐based disulfide bond‐containing biodegradable cationic lipidoid nanoparticles. We have identified a subset of this library which is effective for protein and mRNA delivery in vitro and in vivo. These lipidoids showed comparable transfection efficacies but much lower cytotoxicities compared to the Lpf2k in vitro. In vivo studies in adult mice demonstrated the successful delivery of genome engineering protein and mRNA molecules in the skeletal muscle (via intramuscular injection), lung and spleen (via intravenous injection), and brain (via lateral ventricle infusion).

Non-viral delivery of the CRISPR/Cas system: DNAversusRNAversusRNP

“…Recently, Xu's group designed a series of bioreducible lipidoids for Cas9 mRNA/sgRNA delivery. [191][192][193][194] For example, BAMEA-O16B, one of the leading lipids, formulated with the helper lipids, DSPE-PEG2000 and Cas9 mRNA/ sgPCSK9 could reduce PCSK9 protein levels in mouse serum to…”

“…The same group also reported a new class of cholesteryl-based bioreducible lipidoids as well as another type that showed effective Cas9 mRNA delivery to human mesenchymal stem cells (hMSCs). 192,194 As an alternative to disulfide bond-containing bioreducible lipids, Zhang et al designed a series of lipid-like compounds containing ester groups for Cas9 mRNA delivery. 195 The study showed that linear ester chains were degraded much faster than branched ester chains and both types demonstrated efficient Cas9 mRNA delivery in vitro and in vivo.…”

Tailoring combinatorial lipid nanoparticles for intracellular delivery of nucleic acids, proteins, and drugs

“…To further expand the library of biodegradable lipids, we further incorporated novel head groups (imidazole and its analogs) 41 and tails (cholesteryl-based and cyclic benzylidene acetal-containing tails) 42 , 43 into the lipid molecule design. Our studies found that these newly developed lipid materials are safe and efficient carriers for the intracellular delivery of mRNA, both in vitro and in vivo in mouse models.…”

“…Even though we and others have shown that by altering the lipid chemistry and LNP formulation composition, certain properties ( e.g. , increase cell and tissue specificity, augmented intracellular transfection efficacy, triggered degradability, improved biocompatibility and reduced toxicity, increased membrane fluidity, and enhanced storage stability) could be introduced to the LNP system, a thorough lipid structure‒activity study is needed to fully understand how the LNP systems interact with biological systems both in cell culture and in vivo 31 , 36 , 37 , 42 , 112 . It is critical to understand how the combinatorial lipids with different head, linker, and tail groups and LNPs with different size, morphology, surface chemistry, and charge properties result in differences in cell transfection efficacy, cytotoxicity, and in vivo biodistribution profiles 124 , 125 , 126 , 127 , 128 .…”

Lipidoid nanoparticles increase ATP uptake into hypoxic brain endothelial cells

“…Onpattro was the first FDA-approved siRNA-LNP drug for the treatment of polyneuropathies caused by hereditary transthyretin amyloidosis (5). LNPs have been extensively studied for the delivery of large molecules like polypeptides and nucleic acids (6)(7)(8)(9)(10)(11), however, their delivery potential for small molecule drugs remains unexplored. We propose that delivery of adenosine triphosphate (ATP) to the brain endothelial cells (BECs) lining the blood-brain barrier (BBB) is an effective strategy to increase the cell viability of BECs post-cerebral ischemia/reperfusion injury (stroke).…”