Selection and Identification of Skeletal-Muscle-Targeted RNA Aptamers

Abstract

Oligonucleotide gene therapy has shown great promise for the treatment of muscular dystrophies. Nevertheless, the selective delivery to affected muscles has shown to be challenging because of their high representation in the body and the high complexity of their cell membranes. Current trials show loss of therapeutic molecules to non-target tissues leading to lower target efficacy. Therefore, strategies that increase uptake efficiency would be particularly compelling. To address this need, we applied a cell-internalization SELEX (Systematic Evolution of Ligands by Exponential Enrichment) approach and identified a skeletal muscle-specific RNA aptamer. A01B RNA aptamer preferentially internalizes in skeletal muscle cells and exhibits decreased affinity for off-target cells. Moreover, this in vitro selected aptamer retained its functionality in vivo, suggesting a potential new approach for targeting skeletal muscles. Ultimately, this will aid in the development of targeted oligonucleotide therapies against muscular dystrophies.

Long Noncoding RNA lncMUMA Reverses Established Skeletal Muscle Atrophy following Mechanical Unloading

“…However, there is still no lncRNA-based drug brought into clinical trials to date, especially for skeletal muscle atrophy, due to the difficulty in lncRNA modulation in humans and the potential systematic side effect. Tissue- or organ-specific delivery systems, such as muscle-targeted peptide35, 36 and muscle-targeted aptamer, 37 could be helpful for the lncRNA-based therapy to avoid the potential side effect to other tissues and organs. Another challenge of lncRNA-based therapy is the poor sequence conservation of lncRNA across species.…”

Aptamers Against Live Targets: Is In Vivo SELEX Finally Coming to the Edge?

“…HER2 aptamers and aptamers against myoblasts were selected using this approach. 81,82 Trypsinization can also be used to remove extracellular proteins and, therefore, the bound-aptamer fraction that has not been internalized. 81,83,84 Sgc8 aptamer was selected to bind to human protein tyrosine kinase-7 (PTK7), which is a specific biomarker of acute lymphoblastic leukemia (ALL) T cells.…”

“…81,82 Trypsinization can also be used to remove extracellular proteins and, therefore, the bound-aptamer fraction that has not been internalized. 81,83,84 Sgc8 aptamer was selected to bind to human protein tyrosine kinase-7 (PTK7), which is a specific biomarker of acute lymphoblastic leukemia (ALL) T cells. 85 Other internalizing aptamers against breast cancer cell lines were identified using this cell-SELEX variant.…”

A Bottom-Up Approach for Developing Aptasensors for Abused Drugs: Biosensors in Forensics

“…Finally, the resulting pool is sequenced, and the selected aptamers are characterized for their binding affinity. Since the first introduction of the SELEX method in 1990 by two independent groups [24,25], a large number of aptamers have been developed for various targets ranging from small molecules [26,27] to proteins [28,29], whole cells [30], and tissues [31,32].…”