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tRNA therapeutics for genetic diseases

Abstract

Transfer RNAs (tRNAs) have a crucial role in protein synthesis, and in recent years, their therapeutic potential for the treatment of genetic diseases — primarily those associated with a mutation altering mRNA translation — has gained significant attention. Engineering tRNAs to readthrough nonsense mutation-associated premature termination of mRNA translation can restore protein synthesis and function. In addition, supplementation of natural tRNAs can counteract effects of missense mutations in proteins crucial for tRNA biogenesis and function in translation. This Review will present advances in the development of tRNA therapeutics with high activity and safety in vivo and discuss different formulation approaches for single or chronic treatment modalities. The field of tRNA therapeutics is still in its early stages, and a series of challenges related to tRNA efficacy and stability in vivo, delivery systems with tissue-specific tropism, and safe and efficient manufacturing need to be addressed.

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Fig. 1: Timeline of some key milestones fostering the development of tRNA therapeutics.
Fig. 2: tRNA-based therapeutic modalities.
Fig. 3: Frequency of nonsense mutations in the human population associated with various genetic diseases.
Fig. 4: Therapeutic tRNA delivery approaches.

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Acknowledgements

We acknowledge the helpful discussions with H.-Q. Mao (Institute for NanoBioTechnology, Johns Hopkins University), and P. Kaplan and P. Eimon (Tevard Biosciences). Work in the Coller Lab is supported by NIH grant (R35GM144114), Bloomberg Philanthropies and the Syngap Research Fund, and the research of Z.I. is supported by grants of the Deutsche Forschungsgemeinschaft DFG (IG 73/21-1), NIH (1R01HL136414-05) and Cystic Fibrosis Foundation (IGNATO2010).

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The authors contributed equally to this work.

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Correspondence to Jeff Coller or Zoya Ignatova.

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Competing interests

Z.I. is co-inventor on several licensed and non-licensed patents related to the use of tRNAs as therapeutics and sup-tRNA design and is a member of the scientific advisory board of Tevard Biosciences. J.C. is scientific co-founder of Tevard Biosciences and a member of their scientific advisory board.

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Glossary

Adeno-associated virus

(AAV). Non-pathogenic, small single-stranded DNA virus, whose genome (4.7 kb) encodes four non-structural rep proteins, three capsid (cap) proteins and assembly-activating protein, flanked by two AAV-specific palindromic ITRs (145 bp).

Aminoacyl-tRNA synthetase

(AARS). A universal enzyme family that aminoacylates tRNAs with their cognate amino acid.

Basket trials

Also known as bucket trials; a type of clinical trial for patients with different diseases with the same mutation or biomarker.

Episomal expression

A non-integrated extrachromosomal circular DNA from a viral genome that replicates and is transcribed independently in the eukaryotic nucleus.

Missense mutation

A genetic alteration within the protein-coding sequence leading to a change of the encoded amino acid which may alter the function of a protein.

Natural suppressor tRNAs

Native suppressors of nonsense mutations (mostly in bacteria and yeast) arise from mutation in sense codon decoding tRNA genes enabling the mutant tRNA to translate a stop codon.

Nonsense-mediated mRNA decay

(NMD). A pathway that rapidly degrades mRNA in response to nonsense mutations, which can arise owing to errors in transcription or failure to remove intronic regions, altering the natural reading frame.

Nonsense mutation

A genetic alteration within the protein-coding sequence exchanging a sense codon (that is, encoding an amino acid) to a termination or stop codon, resulting in a loss of protein function.

Recombinant AAV

(rAAV). Engineered AAV, in which the DNA of interest replaces the viral sequences encoding for rep and cap genes, whereas both cis-packaging ITR signals are retained.

Termination codons

Also known as stop codons. Three codons, named amber (UAG), ochre (UAA) and opal (UGA), terminate mRNA translation and, through pairing with the release factor (eRF1 in eukaryotes), release the newly synthesized protein.

Therapeutic threshold

The therapeutic threshold, which is established for each disease based on calculations, descriptive methods and clinical practice, describes the probability of disease at which the condition between treatment and no treatment is the same.

Toll-like receptors

Mediators of inflammatory pathways mediating the immune response towards a variety of pathogen-derived ligands, including DNA, double-stranded RNA, single-stranded RNA and oligonucleotides.

tRNA isoacceptor family

A group of all tRNA isoacceptors carrying the same amino acid.

tRNA isoacceptors

Different tRNA species, which are aminoacylated with the same amino acid, but differ in their anticodon sequences.

tRNA isodecoders

tRNA species that are aminoacylated with the same amino acids and bear the same anticodon, but differ elsewhere in their sequences.

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Coller, J., Ignatova, Z. tRNA therapeutics for genetic diseases. Nat Rev Drug Discov 23, 108–125 (2024). https://doi.org/10.1038/s41573-023-00829-9

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