Gene Therapy https://doi.org/10.1038/s41434-019-0113-4 ARTICLE Neonatal nonviral gene editing with the CRISPR/Cas9 system improves some cardiovascular, respiratory, and bone disease features of the mucopolysaccharidosis I phenotype in mice Roselena Silvestri Schuh 1,2 ● Esteban Alberto Gonzalez 1,3 ● Angela Maria Vicente Tavares 1,4 ● Bruna Gazzi Seolin 4 ● Lais de Souza Elias 1 ● Luisa Natalia Pimentel Vera 1,3 ● Francyne Kubaski 3 ● Edina Poletto 1,3 ● Roberto Giugliani 1,3 ● Helder Ferreira Teixeira 2 ● Ursula Matte 1,3 ● Guilherme Baldo 1,3,4 Received: 30 October 2018 / Revised: 9 October 2019 / Accepted: 19 November 2019 © The Author(s), under exclusive licence to Springer Nature Limited 2019 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 neuroinflamma- tion 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. Introduction Mucopolysaccharidosis type I (MPS I) is an autosomal recessive disease caused by deficiency of the lysosomal enzyme alpha-L-iduronidase (IDUA, EC 3.2.1.76), which is involved in the catabolism of the glycosaminoglycans (GAGs) heparan and dermatan sulfate (DS). Enzyme replacement therapy (ERT) and hematopoietic stem cell transplantation (HSCT) are the two treatments cur- rently available for MPS I. However, these therapies are not completely effective, as ERT is not capable of crossing the blood–brain barrier (BBB) and reach the brain, joints, heart valves, or bones, while HSCT shows to be mostly effective if performed before cognitive decline [1–3]. MPS I clinical spectrum varies from the severe Hurler syndrome (OMIM #67014) to the attenuated Scheie syn- drome (OMIM #67016), with intermediate disease pheno- types classified as Hurler–Scheie syndrome (OMIM#67015) [1, 4]. Multisystemic manifestations as organomegaly, corneal clouding, heart and valve diseases, pulmonary * Ursula Matte umatte@hcpa.edu.br 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 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 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 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 Supplementary information The online version of this article (https:// doi.org/10.1038/s41434-019-0113-4) contains supplementary material, which is available to authorized users. 1234567890();,: 1234567890();,: