A Nanoparticle Platform for Accelerated In Vivo Oral Delivery Screening of Nucleic Acids

Abstract

Oral administration of nucleic acids, such as small interfering RNAs and antisense oligonucleotides, represents a potent modality for treating many gastrointestinal (GI) diseases. However, their use is limited due to a lack of carriers that can effectively mediate their delivery through GI barriers, including gastric enzymes and mucus membranes. Lipid nanoparticles (LNPs) are an emerging delivery system that can protect nucleic acids from degradation and mediate their intracellular delivery, but few studies have evaluated their ability to overcome oral delivery barriers. Here, a high‐throughput in vivo screening platform is developed to accelerate the design of LNPs for oral delivery of nucleic acids. A library of LNPs that encapsulate DNA barcodes (b‐DNAs) was pooled and orally administered to mice, and their delivery was quantified by deep sequencing. Sequencing results indicated that increases in the molar ratio of cholesterol to lipid‐anchored polyethylene glycol conjugate in LNPs enhanced their GI content retention and that the presence of an LNP carrier improved b‐DNA delivery to GI tissues, relative to the delivery of naked b‐DNA. Collectively, these results suggest that high‐throughput in vivo screening can accelerate the discovery of LNPs for GI tract nucleic acid delivery upon oral administration.

Nanoscale delivery platforms for RNA therapeutics: Challenges and the current state of the art

“…127 This approach has been used to directly probe the in vivo behavior of NPs at a throughput that was previously impossible and has resulted in a wealth of data on how NPs are trafficked in vivo. 126,128,129 Of note, distribution of a NP to a certain tissue does not necessarily lead to its expression there. 130,131 To address this issue, Sago et al developed a hybrid approach that combines the Cre mRNA-Ai14 reporter system 132 with DNA barcoding to track functional delivery instead of biodistribution.…”

Oral Delivery of Nucleic Acid Therapies for Local and Systemic Action

“…Moreover, NAs are better drug candidates compared to small molecules for treatment of cancer and rare diseases due to their high degree of selectivity and minimal toxic effects to other tissues. Despite the advantages, delivering NAs orally has numerous challenges discussed at length in this review [ 89 ]. The inherent nature of NAs such as large molecular weight and negative charge combined with physiological aspects like extreme pH in GIT, enzymatic degradation, mucosal layer barrier and peristalsis pose additional challenges for their successful oral delivery [ 90 ].…”

Nucleic acid strategies for infectious disease treatments: The nanoparticle-based oral delivery route

“…In 1999, a DNA-based NP system was developed, whereas DNA encoding for the peanut allergen gene has been shown to transduce gene translation in the epithelium of mucosal and induce systemic immunity upon oral administration of this DNA vaccine ( Li et al, 1999 ). However, the challenge of oral delivery is that various degrading enzymes in the intestine may lead to insufficient antigen uptake and low transfection efficiency ( El‐Mayta et al, 2021 ).…”