2. Academic Validation
  3. Engineered tRNAs suppress nonsense mutations in cells and in vivo

Engineered tRNAs suppress nonsense mutations in cells and in vivo

  • Nature. 2023 May 31. doi: 10.1038/s41586-023-06133-1.
Suki Albers 1,  Elizabeth C Allen 2,  Nikhil Bharti 1,  Marcos Davyt 1,  Disha Joshi 3 4,  Carlos G Perez-Garcia 2,  Leonardo Santos 1,  Rajesh Mukthavaram 2,  Miguel Angel Delgado-Toscano 1,  Brandon Molina 2,  Kristen Kuakini 2,  Maher Alayyoubi 2,  Kyoung-Joo Jenny Park 2,  Grishma Acharya 2,  Jose A Gonzalez 2,  Amit Sagi 2,  Susan E Birket 5,  Guillermo J Tearney 6 7,  Steven M Rowe 5,  Candela Manfredi 3 4,  Jeong S Hong 3 4,  Kiyoshi Tachikawa 2,  Priya Karmali 2,  Daiki Matsuda 2,  Eric J Sorscher 8 9,  Pad Chivukula 10,  Zoya Ignatova 11
Affiliations

Affiliations

  • 1 Institute of Biochemistry and Molecular Biology, University of Hamburg, Hamburg, Germany.
  • 2 Arcturus Therapeutics, San Diego, CA, USA.
  • 3 Department of Pediatrics, School of Medicine, Emory University, Atlanta, GA, USA.
  • 4 Children's Healthcare of Atlanta, Atlanta, GA, USA.
  • 5 Pulmonary, Allergy, and Critical Care Medicine, University of Alabama at Birmingham, Birmingham, AL, USA.
  • 6 Wellman Center for Photomedicine, Massachusetts General Hospital, Boston, MA, USA.
  • 7 Harvard-MIT Health Sciences and Technology, MA, Cambridge, USA.
  • 8 Department of Pediatrics, School of Medicine, Emory University, Atlanta, GA, USA. esorscher@emory.edu.
  • 9 Children's Healthcare of Atlanta, Atlanta, GA, USA. esorscher@emory.edu.
  • 10 Arcturus Therapeutics, San Diego, CA, USA. pad@arcturusrx.com.
  • 11 Institute of Biochemistry and Molecular Biology, University of Hamburg, Hamburg, Germany. zoya.ignatova@uni-hamburg.de.
PMID: 37258671 DOI: 10.1038/s41586-023-06133-1
Abstract

Nonsense mutations are the underlying cause of approximately 11% of all inherited genetic diseases1. Nonsense mutations convert a sense codon that is decoded by tRNA into a premature termination codon (PTC), resulting in an abrupt termination of translation. One strategy to suppress nonsense mutations is to use natural tRNAs with altered anticodons to base-pair to the newly emerged PTC and promote translation2-7. However, tRNA-based gene therapy has not yielded an optimal combination of clinical efficacy and safety and there is presently no treatment for individuals with nonsense mutations. Here we introduce a strategy based on altering native tRNAs into efficient suppressor tRNAs (sup-tRNAs) by individually fine-tuning their sequence to the physico-chemical properties of the amino acid that they carry. Intravenous and intratracheal lipid nanoparticle (LNP) administration of sup-tRNA in mice restored the production of functional proteins with nonsense mutations. LNP-sup-tRNA formulations caused no discernible readthrough at endogenous native stop codons, as determined by ribosome profiling. At clinically important PTCs in the cystic fibrosis transmembrane conductance regulator gene (CFTR), the sup-tRNAs re-established expression and function in cell systems and patient-derived nasal epithelia and restored airway volume homeostasis. These results provide a framework for the development of tRNA-based therapies with a high molecular safety profile and high efficacy in targeted PTC suppression.

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