Challenges in Optimizing RNA Nanostructures for Large-Scale Production and Controlled Therapeutic Properties

Abstract

Nucleic acids have been utilized to construct an expansive collection of nanoarchitectures varying in design, physicochemical properties, cellular processing and biomedical applications. However, the broader therapeutic adaptation of nucleic acid nanoassemblies in general, and RNA-based nanoparticles in particular, have faced several challenges in moving towards (pre)clinical settings. For one, the large-batch synthesis of nucleic acids is still under development, with multi-stranded and chemically modified assemblies requiring greater production capacity while maintaining consistent medical-grade outputs. Furthermore, the unknown immunostimulation by these nanomaterials poses additional challenges, necessary to be overcome for optimizing future development of clinically approved RNA nanoparticles.

The immunorecognition, subcellular compartmentalization, and physicochemical properties of nucleic acid nanoparticles can be controlled by composition modification

“…NANPs can be engineered to perform a variety of therapeutic and diagnostic functions due to the natural biological roles of RNA and DNA ( 1 , 2 , 11 , 15 , 31 , 32 ). However, the transition of NANPs to clinical use has been limited due to issues regarding production, chemical instability, intracellular delivery and off-target stimulation of the immune system ( 12 , 33 , 34 ).…”

“…The NANP panel used in these studies was designed to self-assemble in one-pot with high batch-to-batch consistency due to formation of either A-form or B-form helices via canonical Watson-Crick interactions ( 3 ), which has the potential to become cost effective and advantageous should future scaled-up production be warranted ( 15 ). We constructed seven triangular NANPs with differing compositions shown in Figure 1 : all RNA (RcR), RNA center and DNA sides (RcD), RNA center and 2′F U/C modified RNA sides (Rc2′F), all DNA (DcD), DNA center and RNA sides (DcR), DNA center and 2′F U/C modified RNA sides (Dc2′F), and a fully 2′F U/C modified (2′Fc2′F) RNA triangles.…”

In Vivo Production of RNA Aptamers and Nanoparticles: Problems and Prospects

“…A series of publications devoted to the use of RNA nanoparticles demonstrates the positive experience of using nanoparticles with appropriate ligands for the treatment of cancer. The studies indicate that RNA nanoparticles are characterized by chemical and mechanical stability and can be assembled in vitro from modular blocks, which simplifies their quality control (see reviews by Haque et al; Rossetti et al) [ 68 , 146 ].…”

Drug delivery systems for RNA therapeutics

“…To overcome the barriers to safe and effective RNA delivery, scientists have developed both viral-vector-based and non-viral delivery systems that protect the RNA from degradation, maximize delivery to on-target cells and minimize exposure to off-target cells. Viral gene therapies 3 have generated successful clinical readouts 4 – 9 , but the effectiveness of these approaches can be limited by pre-existing immunity 10 , viral-induced immunogenicity 11 , unwanted genomic integration 12 , payload size constraints 13 , the inability to re-dose, complications involved in upscaling 14 , and expensive vector production. Although scientists are overcoming some of these limitations 15 , they have fuelled the search for alternative drug delivery vehicles.…”