Schejtman Borisonik, Andrea;
(2023)
Lentiviral gene therapy for p47-deficient Chronic Granulomatous disease.
Doctoral thesis (Ph.D), UCL (University College London).
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Abstract
Chronic Granulomatous Disease (CGD) is an inherited primary immunodeficiency disorder with an incidence of ∼1:200,000 live births. This disease is caused by mutations in any of the genes encoding subunits of the NAPDH oxidase, the enzyme responsible for pathogen killing. Mutations in the p47 subunit of the NADPH oxidase cause the most common form of autosomal recessive CGD, a disorder amenable to haematopoietic stem cell (HSC) gene therapy. To this aim, I have developed and tested in animal models a self-inactivating lentiviral vector containing a codon optimized p47phox transgene under the transcriptional control of a myeloid promoter (pCCLChim-p47). The pCCLChim-p47 vector was able to induce high expression of the p47phox protein and restoration of NADPH-oxidase in all models tested. In gene therapy-treated mice levels of NADPH-oxidase activity were comparable to those found in WT animals. The percentage of functional neutrophils remained stable over time in primary and secondary transplants, suggesting that the vector is not prone to epigenetic inactivation. In addition, gene corrected animals were protected against the Salmonella pathogen with a reduction of CFU in the organs analyzed compared to KO and Mock mice. Furthermore, gene transfer with the test vector pCCLChim-p47 had no negative impact on the viability and proliferation of lineage negative cells, suggesting the absence of a general vector- or transgene-related toxicity. Transduction with the vector of human and mouse HSCs did not show adverse effects in the animals. Probing that the lentiviral vector pCCLChim-p47 has a very low in vitro and in vivo genotoxic risk, even at high vector copies per cell. Overall, our experiments paved the way to a clinical development of the pCCLChimp47 vector.
| Type: | Thesis (Doctoral) |
|---|---|
| Qualification: | Ph.D |
| Title: | Lentiviral gene therapy for p47-deficient Chronic Granulomatous disease |
| Open access status: | An open access version is available from UCL Discovery |
| Language: | English |
| Additional information: | Copyright © The Author 2022. Original content in this thesis is licensed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0) Licence (https://creativecommons.org/licenses/by-nc/4.0/). Any third-party copyright material present remains the property of its respective owner(s) and is licensed under its existing terms. Access may initially be restricted at the author’s request. |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10165959 |
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