Cargando…
Development of a fibrin-mediated gene delivery system for the treatment of cystinosis via design of experiment
Cystinosis is a rare disease, caused by a mutation in the gene cystinosin and characterised by the accumulation of cystine crystals. Advantages of biomaterial-mediated gene delivery include reduced safety concerns and the possibility to cure organs that are difficult to treat using systemic gene tra...
Autor principal: | |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8904479/ https://www.ncbi.nlm.nih.gov/pubmed/35260693 http://dx.doi.org/10.1038/s41598-022-07750-y |
_version_ | 1784664959667404800 |
---|---|
author | Graceffa, Valeria |
author_facet | Graceffa, Valeria |
author_sort | Graceffa, Valeria |
collection | PubMed |
description | Cystinosis is a rare disease, caused by a mutation in the gene cystinosin and characterised by the accumulation of cystine crystals. Advantages of biomaterial-mediated gene delivery include reduced safety concerns and the possibility to cure organs that are difficult to treat using systemic gene transfer methods. This study developed novel fibrin hydrogels for controlled, localised gene delivery, for the treatment of cystinosis. In the first part, fabrication parameters (i.e., DNA, thrombin, and aprotinin concentrations) were optimised, using a Design of Experiment (DOE) methodology. DOE is a statistical engineering approach to process optimisation, which increases experimental efficiency, reduces the number of experiments, takes into consideration interactions between different parameters, and allows the creation of predictive models. This study demonstrated the utility of DOE to the development of gene delivery constructs. In the second part of the study, primary fibroblasts from a patient with cystinosis were seeded on the biomaterials. Seeded cells expressed the recombinant CTNS and showed a decrease in cystine content. Furthermore, conditioned media contained functional copies of the recombinant CTNS. These were taken up by monolayer cultures of non-transfected cells. This study described a methodology to develop gene delivery constructs by using a DOE approach and ultimately provided new insights into the treatment of cystinosis. |
format | Online Article Text |
id | pubmed-8904479 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-89044792022-03-09 Development of a fibrin-mediated gene delivery system for the treatment of cystinosis via design of experiment Graceffa, Valeria Sci Rep Article Cystinosis is a rare disease, caused by a mutation in the gene cystinosin and characterised by the accumulation of cystine crystals. Advantages of biomaterial-mediated gene delivery include reduced safety concerns and the possibility to cure organs that are difficult to treat using systemic gene transfer methods. This study developed novel fibrin hydrogels for controlled, localised gene delivery, for the treatment of cystinosis. In the first part, fabrication parameters (i.e., DNA, thrombin, and aprotinin concentrations) were optimised, using a Design of Experiment (DOE) methodology. DOE is a statistical engineering approach to process optimisation, which increases experimental efficiency, reduces the number of experiments, takes into consideration interactions between different parameters, and allows the creation of predictive models. This study demonstrated the utility of DOE to the development of gene delivery constructs. In the second part of the study, primary fibroblasts from a patient with cystinosis were seeded on the biomaterials. Seeded cells expressed the recombinant CTNS and showed a decrease in cystine content. Furthermore, conditioned media contained functional copies of the recombinant CTNS. These were taken up by monolayer cultures of non-transfected cells. This study described a methodology to develop gene delivery constructs by using a DOE approach and ultimately provided new insights into the treatment of cystinosis. Nature Publishing Group UK 2022-03-08 /pmc/articles/PMC8904479/ /pubmed/35260693 http://dx.doi.org/10.1038/s41598-022-07750-y Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Graceffa, Valeria Development of a fibrin-mediated gene delivery system for the treatment of cystinosis via design of experiment |
title | Development of a fibrin-mediated gene delivery system for the treatment of cystinosis via design of experiment |
title_full | Development of a fibrin-mediated gene delivery system for the treatment of cystinosis via design of experiment |
title_fullStr | Development of a fibrin-mediated gene delivery system for the treatment of cystinosis via design of experiment |
title_full_unstemmed | Development of a fibrin-mediated gene delivery system for the treatment of cystinosis via design of experiment |
title_short | Development of a fibrin-mediated gene delivery system for the treatment of cystinosis via design of experiment |
title_sort | development of a fibrin-mediated gene delivery system for the treatment of cystinosis via design of experiment |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8904479/ https://www.ncbi.nlm.nih.gov/pubmed/35260693 http://dx.doi.org/10.1038/s41598-022-07750-y |
work_keys_str_mv | AT graceffavaleria developmentofafibrinmediatedgenedeliverysystemforthetreatmentofcystinosisviadesignofexperiment |