In Vivo Introduction of mRNA Encapsulated in Lipid Nanoparticles to Brain Neuronal Cells and Astrocytes via Intracerebroventricular Administration

Abstract

Gene therapy is a promising strategy for curing certain types of brain diseases. Supplementation of therapeutic proteins such as aromatic amino acid decarboxylase (AADC) or nerve growth factor (NGF) have been reported to be successful examples of such treatments. However, there are safety concerns because these systems are based on virus-based gene vectors. A safe and efficient artificial carrier is thus urgently needed as an alternative. In this study, an mRNA based artificial gene carrier was introduced into the mouse brain via intracerebroventricular administration. As a carrier, a lipid nanoparticle (LNP) composed of environmentally sensitive lipid-like materials called an SS-cleavable proton-activated lipid-like material is used. The apolipoprotein E mediated cellular uptake of the lipid nanoparticles is one of the key features for its superior and homogeneous transfection activity compared to commercially available transfection reagents in both in vitro and in vivo situations. Immunostaining of brain specimens suggested that exogenous proteins can be introduced into neuronal cells as well as astrocytes using the mRNA-based gene carrier. This cannot be achieved using DNA-based artificial gene carriers. The findings suggest that a combination of an mRNA and a lipid based delivery system have great promise as a platform for the treatment of brain disorders.

Gene therapy for neurodegenerative disorders: advances, insights and prospects

“…Moreover, pseudotyping lentiviruses with glycoproteins were found to selectively transfect astrocytes after intraparenchymal administration 150 , 151 . Nanoparticles functionalized with bradykinin B2 receptor antibodies, transferrin receptor or apolipoprotein E have also been indicated to successfully deliver siRNA and mRNA to astrocytes 152 , 153 . For example, optimized branched poly ( β -amino ester)s are applied to deliver NGF expression DNA to astrocytes, and high transfection efficiency is achieved, which provides a viable gene therapy approach for neurodegenerative disorders 154 .…”

Inhibition of the Inflammatory Pathway Enhances Both the <i>in Vitro</i> and <i>in Vivo</i> Transfection Activity of Exogenous <i>in Vitro</i>-Transcribed mRNAs Delivered by Lipid Nanoparticles

“…We developed a series of ionizable lipids (SS-cleavable and pH-activated lipid-like material; ssPalm) that is equipped with hydrophobic scaffolds, proton-sponging tertiary amines and cleavable disulfide bonding for achieving the cytoplasmic delivery of nucleic acids. [1][2][3] The ssPalm can form an mRNAencapsulating lipid nanoparticle that possesses a neutral surface charge in a physiological environment (LNP ssPalm ). Once taken up by cells, the LNP ssPalm develops a positive charge via protonation of the tertiary amines in the acidic endosomal compartments.…”

Gold nanorods/siRNA complex administration for knockdown of PARP-1: a potential treatment for perinatal asphyxia

“…66 The positive charge triggers an interaction between the carrier and the endosomal membrane for endosomal escape. [67][68][69] However, the exact mechanism of endosomal escape of many siRNA delivery carriers is still poorly understood. 70 For in vivo characterization, we used a model of global PA, 2 administering the complex in a single dose (i.p.)…”