mRNA‐Loaded Lipid‐Like Nanoparticles for Liver Base Editing Via the Optimization of Central Composite Design

Abstract

Messenger RNA (mRNA) has come into the spotlight due to its potential for addressing a staggering number of diseases, while the lack of effective and safe carriers for in vivo delivery significantly limits its clinical application. Herein, the potential of lipid‐like nanoparticles (LLNs) containing three new cholesterol derivatives to achieve the liver targeting delivery of mRNA is investigated. The central composite design (CCD) is used to tailor the formulation of LLNs through in vivo optimization of the molar ratios of these cholesterol derivatives required for liver targeting. The optimized LLNs (O‐LLNs) are able to systemically deliver mRNA to the liver of mice with a synergistic action of prolonged systemic circulation, increased liver targeting, and enhanced hepatocyte uptake. The O‐LLNs outperformed DLin‐MC3‐DMA (MC3) in functional delivery of Cre‐recombinase (Cre) and human erythropoietin (hEPO) mRNA. Successful delivery of cytidine base editor mRNA (CBE mRNA) and sgRNA by O‐LLNs achieved more than 8% correction rates in a liver‐related metabolism disorder, phenylketonuria (PKU). In conclusion, the general method above can accelerate in vivo screening and optimization of multicomponent nanoparticle formulations, and the optimized ones demonstrate great potential as delivery vehicles for targeted gene therapy in specific tissues.

PEGylated pH-responsive peptide-mRNA nano self-assemblies enhance the pulmonary delivery efficiency and safety of aerosolized mRNA

“…A549 is a human alveolar epithelial adenocarcinoma cell line commonly used to evaluate mRNA delivery agents’ performance through intranasal or pulmonary administration (Qiu et al., 2021 ). HepG2 is a representative human hepatocellular carcinomas cell line that may influence mRNA metabolism (Zheng et al., 2021 ). As shown in Figure 3(A) and (B) , the cytotoxicity of PEGylated peptides (PEG 12 LA and PEG 24 LA) were significantly lower than the non-PEGylated LA peptide at high concentrations (50 and 25 μg/mL), indicating PEGylation could decrease the vector’s cytotoxicity in vitro .…”

Advances in nanoparticle-based mRNA delivery for liver cancer and liver-associated infectious diseases

“…By utilizing the mannose receptors on LSECs, mannose-modified lipid nanoparticles have shown to accumulate in the liver in an in vivo study. 88 The method of administration of mRNA can also affect the uptake of mRNA in the liver. Hydrodynamic delivery is a rapid injection of genetic material to alter the hydrodynamic pressure in capillaries to increase cell permeability temporarily.…”

mRNA-based therapeutics: powerful and versatile tools to combat diseases

“…Our lab has been committed to promoting mRNA-based therapeutics to become powerful and versatile tools to combat diseases, especially in gene therapy and immunotherapy. 14 We have developed diverse novel targeted delivery nanoparticles 15 and constructed receptor-binding domain (RBD)-encoding mRNA formulated in liposomes to prevent and treat the SARS-CoV-2 pandemic. 16 In this review, we comprehensively summarize the recent progress towards mRNA design and synthesis, as well as the enabling of mRNA delivery technologies.…”