Benoist, Thomas;
(2024)
Engineering vascularised alveolar epithelium.
Doctoral thesis (Ph.D), UCL (University College London).
Abstract
The development of engineered human lung tissue could bring a solution to various current medical problems. First, it could help bring a more representative alternative to animal models for testing human disease and treatment. Pre-clinical testing of therapy candidates onto human tissues and/or disease models could indeed help establish their efficacy and safety profiles in a more precise way, especially when inter-species differences in disease pathophysiology and drug response are involved. Then, engineered lung tissues could be used medically for transplantation, potentially alleviating the various problems associated to allogeneic lung transplantation: namely the scarcity of donor lungs, the necessity of a lifelong immunosuppressant therapy and the occurrence of graft failure within 10 years. Therefore, to develop engineered lung tissue, we first optimised an existing lung engineering bioreactor and used it to decellularise and recellularise whole rodent lungs. Then, we assessed the biological relevance of this engineered lung tissue by using it as a disease model and submitting it to respiratory syncytial virus (RSV) infection and antiviral treatment, using an adapted version of our bioreactor setup. After 4 days of dynamic culture, engineered lungs showed multiple signs of RSV infection and had their phenotype rescued by antiviral treatment. Finally, we tried to improve engineered lung tissue maturation, especially on the alveolar side, by providing various physiological stimuli. First, we successfully optimised a culture system to vascularise alveolar epithelial organoids with reprogrammed vascular endothelial cells (rVECs), which upregulated the alveolar type 1 (AT1) epithelial cell marker AQP5 (aquaporin 5) (p<0.05), suggesting that it may promote tissue differentiation. Then, perfusion of rVECs within specialized culture systems allowed the formation of lumenised and perfusable vascular networks. Finally, I designed a novel bioreactor setup for stretching human lung bud organoid (hLBO)-seeded lung extracellular matrix (ECM) slices. After 7 days of stretching, quantification of immunofluorescence results showed significantly increased alveolar type 2 (AT2) epithelial cell numbers in stretched versus unstretched tissue (p<0.05). We were therefore able to produce biologically relevant engineered lung tissue and find steps to improve the maturation of its alveolar compartment.
| Type: | Thesis (Doctoral) |
|---|---|
| Qualification: | Ph.D |
| Title: | Engineering vascularised alveolar epithelium |
| Language: | English |
| UCL classification: | 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 > 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/10192490 |
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