UCL Discovery
UCL home » Library Services » Electronic resources » UCL Discovery

Towards gene therapy for primary ciliary dyskinesia

Munye, MM; (2014) Towards gene therapy for primary ciliary dyskinesia. Doctoral thesis , UCL (University College London). Green open access

[thumbnail of Thesis FINAL submitted.pdf]
Preview
Text
Thesis FINAL submitted.pdf
Available under License : See the attached licence file.

Download (5MB)

Abstract

Primary ciliary dyskinesia is a genetic disorder where patients develop lung disease as they are unable to clear airway infections effectively. There is currently no treatment for the underlying genetic defect. This thesis describes advances towards the development of a gene therapy targeting the airway disease in PCD patients with DNAH5 mutations, the most common cause of PCD. Little work has been done in the field so many challenges remain. DNAH5 is a large gene and the cDNA has not been cloned. In addition PCD models are currently inadequate. Finally, non-viral vectors for airway gene delivery produce sub-optimal levels of transgene expression and more efficient viral vectors cannot package the large DNAH5 cDNA. To address these issues functional DNAH5 cDNA was cloned from healthy airway cells and the cDNA validated by sequencing and expression studies. Lentiviral shRNA transduction of healthy cells was used to knock down DNAH5 in airway cells semi-immortalised with BMI-1, a proto-oncogene that allows extended growth capacity of cells whilst retaining their mucociliary differentiation potential. The novel cell lines lacked cilia motility as is seen in patient cells. Minicircle DNA, in the context of airway gene delivery, was found to enhance gene expression in vitro and in vivo. A non-viral vector was optimised, characterised and used to deliver DNAH5 coding minicircle DNA to PCD models but poor transfection efficiency of DNAH5 prevented functional correction. Transfection of smaller genes was efficient so the vector in its current form could be useful for gene therapy treatment of the majority of PCD causing genes. Studies are needed to determine and overcome the bottlenecks in the efficient transfection of large transgenes to help advance PCD gene therapy.

Type: Thesis (Doctoral)
Title: Towards gene therapy for primary ciliary dyskinesia
Open access status: An open access version is available from UCL Discovery
Language: English
UCL classification: UCL > Provost and Vice Provost Offices
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/1435816
Downloads since deposit
706Downloads
Download activity - last month
Download activity - last 12 months
Downloads by country - last 12 months

Archive Staff Only

View Item View Item