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UCL Great Ormond Street Institute of Child Health

Great Ormond Street Institute of Child Health

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Developing novel RNA editing therapy for neuromuscular disorders

Supervisors names
Haiyan Zhou
Jinhong Meng

Background - Duchenne Muscular Dystrophy (DMD) is a rare genetic disorder affecting approximately 1 in 5000 boys at birth. It is characterized by progressive muscle weakness caused by a mutation in the Dystrophin gene, which impairs normal muscle function. In 15-30% of DMD cases, a point mutation leads to a nonsense mutation, resulting in reduced protein production.

Aims/Objectives - To develop an optimal RNA editing approach for correcting the nonsense mutation in DMD muscle cells.

Methods- We will treat DMD muscle cells with gRNA-ADAR delivered by AONs or AAVs, closely monitor the correction of the nonsense mutation and any associated side-effects. By comparing the outcomes between the two delivery methods, we aim to identify the optimal approach that achieves efficient and safe RNA editing.

Once the optimal RNA editing approach is determined, it will be applied to DMD muscle cells derived from induced pluripotent stem cells (iPSCs) to restore the reading-frame. The efficacy of the RNA editing approach will be evaluated in vitro by assessing the expression of Dystrophin after treatment. Furthermore, corrected cells will be transplanted into a DMD mouse model for evaluating the ex vivo cell therapy strategy. Alternatively, we will also inject the optimal AONs or AAVs into a DMD mouse model engrafted with DMD human cells to assess in vivo editing efficiency.

Timeline

  • Months 1-9: Establish cellular platform for evaluation using DMD iPSC Pax7-GFP reporter cell lines.
  • Months 1-12: Design and validate AONs or AAVs in HEK293 cells.
  • Months 13-24: Evaluate RNA editing efficiency in DMD iPSC-derived myoprogenitor cells.
  • Months 25-30: Assess side-effects in treated cells through deep RNA sequencing.
  • Months 31-42: Evaluate in vivo efficiency by injecting optimal AONs or AAVs into partially humanized mdx nude mouse muscle.
  • Months 43-48: Prepare manuscripts and write the thesis.

References

  • Aartsma-Rus, A., I.B.Ginjaar, and K.Bushby. 2016. J. Med. Genet. 53:145-151.
  • Chal, J., T.Z.Al, et al. 2016. Nat. Protoc. 11:1833-1850.
  • Guiraud, S., A.Aartsma-Rus, et al. 2015. Annu. Rev. Genomics Hum. Genet. 16:281-308.
  • Viggiano, E., E.Picillo, et al. 2023. Genes (Basel) 14.
  • Li, G., M.Jin, et al. 2023. J. Clin. Invest 133.

Contact
Haiyan Zhou. haiyan.zhou@ucl.ac.uk