Wantuch, Slawomir;
(2023)
Hematopoietic stem cell gene therapy for Pompe disease using a novel recombinant form of acid-alpha glucosidase.
Doctoral thesis (Ph.D), UCL (University College London).
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Abstract
Glycogen storage disease type II, also known as Pompe disease (PD), is caused by aberrant or absent expression of the lysosomal enzyme acid-alfa-glucosidase (GAA) that is responsible for the breakdown of glycogen into glucose within cells. Babies born with PD suffer from muscle weakness, cardiac myopathy and consequently without treatment they die in the 1st year of life due to respiratory and cardiac failure. In contrast, adolescence PD has a milder course and adverse effects. Currently, patients with PD are treated with enzyme replacement therapy (ERT), where recombinant GAA enzyme is intravenously (i.v.) injected and through bloodstream distributed into lysosomes utilising the mannose-6-phosphate receptor (M6PR). However, ERT is limited by the host humoral response to exogenous recombinant GAA, the lack of penetration the blood-brain-barrier, and finally, an inadequate expression of M6PR on muscle tissues, limiting cross-correction in skeletal muscle. Autologous Hematopoietic Stem Cells (HSCs) Gene Therapy for PD could potentially provide constant and life-long source of endogenous GAA enzyme. Due inadequate expression of M6PRs on skeletal muscle cells limiting cross-correction, this study aims to interrogate alternative approach : using a novel muscle homing peptides (MHPs) (X. Gao et al., 2014; Samoylova & Smith, 1999), which shown strong affinity for skeletal muscle and proving better GAA uptake into these tissues. For this purpose, MHP was incorporated into the N-terminus of GAA. In addition, a synthetic secretion peptide (SP) was placed upstream of MHP to improve secretion of the enzyme from producing cells. In vitro study showed advantageous uptake of MHP-GAA protein to Gaa -/- murine myoblast. The addition of the SP led to 2.5-fold increase of GAA enzymes being secreted from transduced cells in vitro. This project also proposed a cellular receptor involved in the interaction with the MHP. Furthermore, an in vivo evaluation of cross-correction in Pompe mouse model was conducted confirming long-term GAA transgene expression in transduced and transplanted HSCs. Also, cross-correction and reduction of glycogen in harvested tissues in GT treated Gaa -/- mice was observed, further supporting GT as a potential treatment to alleviate Pompe disease symptoms.
| Type: | Thesis (Doctoral) |
|---|---|
| Qualification: | Ph.D |
| Title: | Hematopoietic stem cell gene therapy for Pompe disease using a novel recombinant form of acid-alpha glucosidase |
| Open access status: | An open access version is available from UCL Discovery |
| Language: | English |
| Additional information: | Copyright © The Author 2023. 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/10167394 |
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