Development of in vivo Gene Editing Therapy for severe Mucopolysaccharidosis type I: preclinical approach in hIDUA-MPS I-H murine models using lipid nanoparticles

• Grant number: 23/15326-0
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: August 01, 2024
• End date: February 29, 2028
• Field of knowledge: Biological Sciences - Genetics - Human and Medical Genetics
• Principal Investigator: Ângela Saito
• Grantee: Rodrigo Miranda de Carvalho
• Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovação (Brasil). Campinas , SP, Brazil

Abstract

Mucopolysaccharidosis type I (MPS I) is a monogenic disease, with autosomal recessive inheritance, caused by mutations in the IDUA gene. These mutations result in the absence or deficiency of activity of the lysosomal enzyme alpha-L-Iduronidase (IDUA), necessary for the degradation of iduronic acid residues of glycosaminoglycans. The severe form of MPS I is known as Hurler Syndrome (MPS I-H), which manifests before 2 years of age, leads to multisystem damage, progressive neurodegenerative clinical evolution and death generally before the age of 10 years. The most common mutation of MPS I-H in Brazil and in several countries around the world is p.W402X (c.1206 G>A), and the only available therapy for the severe condition is allogeneic hematopoietic stem cell (HSC) transplantation, however, there are challenges as lack of compatible donors and graft-versus-host disease. Therefore, as it is a serious disease with no effective treatments available, this project aims to develop an innovative in vivo gene editing approach as a curative therapy proposal for MPS I-H caused by the W402X mutation. CRISPR-Cas9 genome editing technologies as well as the enhanced version Prime Editing (PE) will be employed to create the human disease model and correction of the referred mutation, respectively. PE, in addition to its versatility, has demonstrated safer clinical application potential than CRISPR-Cas9 as it does not require direct double-strand cleavage for gene editing. The in vivo delivery approach of PE technology will utilize lipid nanoparticles (LNPs) formulated with PE-RNAs and functionalized with anti-CD117 antibody (CD117/LNP-RNA-PE) to target and deliver the editing machinery to HSCs. Tests will be carried out, initially on bone marrow cells in vitro and in vivo from transgenic mice, aiming to optimize CD117/LNP-RNA conditions, followed by validations and quality controls of the therapeutic proposal in cellular and murine disease models. The procedures developed in this innovative project, for targeted delivery and gene editing of HSCs in vivo, could be applied as a curative therapy not only for MPS I-H, but also for hematopoietic diseases and other neurometabolic conditions with epidemiological relevance in Brazil.