Nasal Administration of Cationic Nanoemulsions as Nucleic Acids Delivery Systems Aiming at Mucopolysaccharidosis Type I Gene Therapy
This study demonstrates the nasal administration (NA) of nanoemulsions complexed with the plasmid encoding for IDUA protein (pIDUA) as an attempt to reach the brain aiming at MPS I gene therapy.
Formulations composed of DOPE, DOTAP, MCT (NE), and DSPE-PEG (NE-PEG) were prepared by high-pressure homogenization, and assessed in vitro on human fibroblasts from MPS I patients and in vivo on MPS I mice for IDUA production and gene expression.
The physicochemical results showed that the presence of DSPE-PEG in the formulations led to smaller and more stable droplets even when submitted to dilution in simulated nasal medium (SNM). In vitro assays showed that pIDUA/NE-PEG complexes were internalized by cells, and led to a 5% significant increase in IDUA activity, besides promoting a two-fold increase in IDUA expression. The NA of pIDUA/NE-PEG complexes to MPS I mice demonstrated the ability to reach the brain, promoting increased IDUA activity and expression in this tissue, as well as in kidney and spleen tissues after treatment. An increase in serum IL-6 was observed after treatment, although with no signs of tissue inflammatory infiltrate according to histopathology and CD68 assessments.
These findings demonstrated that pIDUA/NE-PEG complexes could efficiently increase IDUA activity in vitro and in vivo after NA, and represent a potential treatment for the neurological impairment present in MPS I patients.
KEY WORDSGene therapy mucopolysaccharidosis type I nanoemulsion nasal administration nonviral vectors
Blank cationic nanoemulsions
Blank cationic nanoemulsions containing DSPE-PEG
1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino (polyethylene glycol)-2000]
Medium Chain Triglycerides
Plasmid encoding for IDUA protein
Nanoemulsion complexed with pIDUA
Nanoemulsion containing DSPE-PEG complexed with pIDUA
Simulated nasal medium
ACKNOWLEDGMENTS AND DISCLOSURES
Authors would like to thank the Brazilian National Council for Scientific and Technological Development (CNPq), Foundation for Research Support of the State of Rio Grande do Sul (FAPERGS, #17/2551–0001273-9), and HCPA Foundation for Incentive of Research (FIPE/HCPA, #150215) for the financial support. J. Bidone wishes to thank the Foundation for Research Support of the State of Rio Grande do Sul (DOCFIX/FAPERGS) and R. S. Schuh would like to thank CNPq (#151021/2018–0) for their postdoctoral grant. H. F. Teixeira, G. Baldo, and U. Matte are recipients of CNPq research fellowship. The authors have made substantial contributions to all of the following: R. S. Schuh and J. Bidone contributed to the conception and design of the study. G. Pasqualim, T. G. de Carvalho, É. Poletto, C. V. Pinheiro, D. S. Diel, M. Farinon, and R. M. Xavier contributed to the acquisition of data, analysis, and interpretation of data. G. Baldo, U. Matte, and H. F. Teixeira contributed to drafting the article and revising it critically for important intellectual content. All authors approved the final version to be submitted. All institutional and national guidelines for the care and use of patients’ material were followed and all patients and/or their caregivers gave written informed consent for this study. All institutional and national guidelines for the care and use of laboratory animals were followed (Hospital de Clínicas de Porto Alegre Ethics Committee #15–0215). All authors declare that they have no conflict of interest.
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