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Title: Gene therapy for alpha-1-antitrypsin deficiency
Author: Paul-Smith, Michael Christian
ISNI:       0000 0004 7657 3358
Awarding Body: Imperial College London
Current Institution: Imperial College London
Date of Award: 2017
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Alpha-1-antitrypsin deficiency (AATD) is an autosomal recessive disorder characterised by low levels of the circulating anti-protease alpha-1-antitrypsin (hAAT). Low levels of pulmonary hAAT pre-disposes individuals to early-onset emphysema, and the efficacy of intravenous augmentation therapy with purified protein is the subject of debate. Gene therapy could be an alternative treatment strategy. In this thesis, "gold standard" viral and non-viral agents for airway gene transfer were assessed for their ability to produce alpha-1-antitrypsin. Key findings in this thesis are: 1. A single dose of gene therapy with the non-viral agent GL67A was not sufficient to produce hAAT in murine lung tissue homogenate or epithelial lining fluid (ELF). A repeat dosing regimen resulted in significant (p < 0.01) but sub-therapeutic production of hAAT in ELF (treated: median 331 (range 0.0-1107) ng/ml hAAT, compared to the therapeutic target of 70 µg/ml in ELF, n=6 per group) (Chapter 3). 2. A lentiviral vector, pseudotyped with Sendai virus surface proteins (rSIV.F/HN-hCEF-sohAAT) resulted in therapeutic levels of hAAT in murine ELF (treated: median 70.0 (range 53.8-175.7 µg/ml, untreated: 0.19 (0.12-0.34) µg/ml, p < 0.05, n=6 per group) (Chapter 5). 3. A single dose of rSIV.F/HN resulted in sustained transgene expression for at least 20 months in murine lung and ELF (p < 0.05) (Chapter 5). 4. In an ex vivo assay, broncho-alveolar lavage fluid (BALF) from rSIV.F/HN-treated mice inhibited neutrophil elastase significantly more than BALF from untreated mice (p < 0.05) (Chapter 7). 5. rSIV.F/HN-hCEF-sohAAT successfully transduced human lung tissue ex vivo; precision-cut human lung slices (treated: median 320 (range 207-1307) ng/ml, untreated: 9.3 (7.5-10) ng/ml, p < 0.01, n=6 per group) and primary human nasal epithelial cells (treated: 389 (339-502) ng/ml, untreated: 140 (103-176) ng/ml, p < 0.01, n=6 per group) (Chapter 6). In summary, data generated in this thesis supports further progression of rSIV.F/HN-hCEF-sohAAT towards a clinical trial in hAAT deficient patients.
Supervisor: Griesenbach, Uta ; Alton, Eric Sponsor: Medical Research Council
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral