Airway and Muscle Gene Transfer to Create Therapeutic Protein Factories
Protein based therapeutics, once a rarely used subset of medical treatments, are now widely adopted with over 130 different proteins/peptides approved for clinical use. Hormone and enzyme replacement therapies have had dramatic clinical impact in chronic diseases such as cystic fibrosis, diabetes and the lysosomal storage disorders (LSDs) Pompe and Gauchers disease. However these therapies can be associated with high treatment burdens (e.g. multiple daily inhalations or injections) and extremely high treatment costs (e.g. $200-700K per year for LSDs).
We have been focusing on an alternative strategy to deliver these therapeutics via delivery of an efficient gene transfer vector to either the muscle or respiratory epithelium to generate a “protein factory”; capable of secreting therapeutic proteins into both the lung lumen and the systemic circulation to provide an increased quality of life and a decreased treatment cost for a range of lung disease, endocrine diseases and inborn errors of metabolism.
Our preferred muscle gene delivery vector is recombinant adeno-associated virus (rAAV). Our preferred lung gene delivery vector is a novel, patent protected, third-generation, self-inactivating simian immunodeficiency virus in which the envelope proteins have been replaced with the F & HN proteins from Sendai virus (rSIV.F/HN) to increase airway cell targeting. Delivery of either vector results in abundant and long-lived expression of transgenes: for example, therapeutic monoclonal antibodies can be expressed for the lifetime of experimental animals. This project will utilise our experience of in vivo gene transfer to understand and manipulate the factors required for effective expression and secretion of therapeutic proteins. This information could then be used to develop vectors for a variety of disease-specific applications including expression of: alpha-galactosidase, alpha-glucosidase or beta-glucocerebrosidase for LSDs. Outcomes such as metabolic, haemostatic, muscle and cardio-respiratory function will be evaluated in human cell culture and knockout/genome engineered mouse models.
The project will be based in the Gene Medicine Research Group in the John Radcliffe Hospital. The group are experts in the development of gene therapy for lung diseases, and has experience in conducting clinical gene therapy trials, exposing students to all aspects of translational research. The student will receive training in techniques such as: molecular biology, genome engineering, cell culture, microscopy & imaging, protein characterisation along with virus production/purification and functional evaluation, PCR, FACS, Western blotting, immunocytochemistry, ELISA, Quantitative (RT)-PCR, chromatography, & Tangential Flow Filtration (TFF) methods.
As well as the specific training detailed above, students will have access to a wide-range of seminars and training opportunities through the many research institutes and centres based in Oxford. Students are also able to attend the Methods and Techniques course run by the MRC Weatherall Institute of Molecular Medicine. This course runs through the year, ensuring that students have the opportunity to build a broad-based understanding of differing research techniques.
Generic skills training is offered through the Medical Sciences Division's Skills Training Programme. This programme offers a comprehensive range of courses covering many important areas of researcher development: knowledge and intellectual abilities, personal effectiveness, research governance and organisation, and engagement, influence and impact. Students are actively encouraged to take advantage of the training opportunities available to them.
The Department has a successful mentoring scheme, open to graduate students, which provides an additional possible channel for personal and professional development outside the regular supervisory framework. We hold an Athena SWAN Silver Award in recognition of our efforts to support the careers of female students and staff.
|1||Gill DR, Bazzani RP, Hyde SC. 2010. Strategies for long-term expression of transgenes in the respiratory epithelium.Curr. Opin. Mol. Ther., 12 (4), pp. 386-93. - http://www.ncbi.nlm.nih.gov/pubmed/20677089|
|2||Griesenbach U, Inoue M, Meng C, Farley R, Chan M, Newman NK, Brum A, You J, Kerton A, Shoemark A, Boyd AC, Davies JC, Higgins TE, Gill DR, Hyde SC, Innes JA, Porteous DJ, Hasegawa M, Alton EW. 2012. Assessment of F/HN-pseudotyped lentivirus as a clinically relevant vector for lung gene therapy.Am. J. Respir. Crit. Care Med., 186 (9), pp. 846-56. - http://www.ncbi.nlm.nih.gov/pubmed/22955314|
Alton EW, Beekman JM, Boyd AC, Brand J, Carlon MS, Connolly MM, Chan M, Conlon S, Davidson HE, Davies JC, Davies LA, Dekkers JF, Doherty A, Gea-Sorli S, Gill DR, Griesenbach U, Hasegawa M, Higgins TE, Hironaka T, Hyndman L, McLachlan G, Inoue M, Hyde SC, Innes JA, Maher TM, Moran C, Meng C, Paul-Smith MC, Pringle IA, Pytel KM, Rodriguez-Martinez A, Schmidt AC, Stevenson BJ, Sumner-Jones SG, Toshner R, Tsugumine S, Wasowicz MW, Zhu J.
2017. Preparation for a first-in-man lentivirus trial in patients with cystic fibrosis.
Thorax, Feb;72(2):137-147. - https://www.ncbi.nlm.nih.gov/pubmed/27852956