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Neonatal nonviral gene editing with the CRISPR/Cas9 system improves some cardiovascular, respiratory, and bone disease features of the mucopolysaccharidosis I phenotype in mice

Gene Therapy volume 27, pages 74–84 (2020)Cite this article

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Abstract

Mucopolysaccharidosis type I (MPS I) is caused by deficiency of alpha-L-iduronidase (IDUA), leading to multisystemic accumulation of glycosaminoglycans (GAG). Untreated MPS I patients may die in the first decades of life, mostly due to cardiovascular and respiratory complications. We previously reported that the treatment of newborn MPS I mice with intravenous administration of lipossomal CRISPR/Cas9 complexes carrying the murine Idua gene aiming at the ROSA26 locus resulted in long-lasting IDUA activity and GAG reduction in various tissues. Following this, the present study reports the effects of gene editing in cardiovascular, respiratory, bone, and neurologic functions in MPS I mice. Bone morphology, specifically the width of zygomatic and femoral bones, showed partial improvement. Although heart valves were still thickened, cardiac mass and aortic elastin breaks were reduced, with normalization of aortic diameter. Pulmonary resistance was normalized, suggesting improvement in respiratory function. In contrast, behavioral abnormalities and neuroinflammation still persisted, suggesting deterioration of the neurological functions. The set of results shows that gene editing performed in newborn animals improved some manifestations of the MPS I disorder in bone, respiratory, and cardiovascular systems. However, further studies will be imperative to find better delivery strategies to reach “hard-to-treat” tissues to ensure better systemic and neurological effects.

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Fig. 1: GAG quantification using Tandem Mass Spectrometry.
Fig. 2: Facial morphology and body weight.
Fig. 3: Effect of gene editing on bone abnormalities.
Fig. 4: Left ventricular mass assessed by echocardiography.
Fig. 5: Heart valve thickening and GAG storage.
Fig. 6: Pathology in the aorta.
Fig. 7: Respiratory disease.
Fig. 8: Behavior analysis: open field and inhibitory avoidance test.
Fig. 9: Neuroinflammation.

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Acknowledgements

The authors were supported by National Council for Scientific and Technological Development (CNPq) (grant numbers 470888/2014-8 and 141742/2014-3), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil (CAPES) (Finance Code 001), Universidade Federal do Rio Grande do Sul, FAPERGS (Grant 02/2017—Programa Pesquisador Gaúcho numbers 17/2551-0001103-1 and 17/2551-0001273-9) and FIPE—Hospital de Clínicas de Porto Alegre, Serviço de Pesquisa Experimental (grant number 2015-0416). RSS would like to thank CNPq for the postdoctoral grant (PDJ grant number 151021/2018-0).

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Authors and Affiliations

  1. Centro de Terapia Gênica, Serviço de Pesquisa Experimental - Hospital de Clinicas de Porto Alegre, R. Ramiro Barcelos 2350, 90035-903, Porto Alegre, RS, Brazil

    Roselena Silvestri Schuh, Esteban Alberto Gonzalez, Angela Maria Vicente Tavares, Lais de Souza Elias, Luisa Natalia Pimentel Vera, Edina Poletto, Roberto Giugliani, Ursula Matte & Guilherme Baldo

  2. Programa de Pós-Graduação em Ciências Farmacêuticas da Universidade Federal do Rio Grande do Sul (UFRGS), Faculdade de Farmácia, Av. Ipiranga 2752, 90610-000, Porto Alegre, RS, Brazil

    Roselena Silvestri Schuh & Helder Ferreira Teixeira

  3. Programa de Pós-Graduação em Genética e Biologia Molecular da Universidade Federal do Rio Grande do Sul (UFRGS), Campus do Vale, Av. Bento Gonçalves 9500, 91501-970, Porto Alegre, RS, Brazil

    Esteban Alberto Gonzalez, Luisa Natalia Pimentel Vera, Francyne Kubaski, Edina Poletto, Roberto Giugliani, Ursula Matte & Guilherme Baldo

  4. Programa de Pós-Graduação em Fisiologia da Universidade Federal do Rio Grande do Sul (UFRGS), Instituto de Ciências Básicas da Saúde, R. Sarmento Leite 500, 90035-190, Porto Alegre, RS, Brazil

    Angela Maria Vicente Tavares, Bruna Gazzi Seolin & Guilherme Baldo

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Schuh, R.S., Gonzalez, E.A., Tavares, A.M.V. et al. Neonatal nonviral gene editing with the CRISPR/Cas9 system improves some cardiovascular, respiratory, and bone disease features of the mucopolysaccharidosis I phenotype in mice. Gene Ther 27, 74–84 (2020). https://doi.org/10.1038/s41434-019-0113-4

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