GSBS Dissertations and Theses
Approval Date
8-5-2014
Document Type
Doctoral Dissertation
Academic Program
Interdisciplinary Graduate Program
Department
Neurology
First Thesis Advisor
Miguel Sena-Esteves, Ph.D.
Keywords
Dependovirus, G(M1) Ganglioside, GM1 Gangliosidosis, Genetic Vectors, Lysosomal Storage Diseases, beta-Galactosidase
Subjects
Dissertations, UMMS; Dependovirus; G(M1) Ganglioside; Gangliosidosis, GM1; Genetic Vectors; Lysosomal Storage Diseases; beta-Galactosidase
Abstract
GM1 gangliosidosis is a lysosomal storage disorder caused by a deficiency in the catabolizing enzyme β-galactosidase (βgal). This leads to accumulation of GM1-ganglioside (GM1) in the lysosome inducing ER stress and cell death. GM1 gangliosidosis is primarily a disorder of the central nervous system (CNS) with peripheral organ involvement. In this work we report two major findings, 1) systemic treatment of GM1 gangliosidosis with an adenoassociated virus (AAV9) encoding mouse-βgal (mβgal) in a GM1 gangliosidosis mouse model (βGal-/-), and 2) an investigation into an intracranial injection of a therapeutic AAVrh8 encoding mβgal. Systemic treatment of GM1 gangliosidosis with AAV9 resulted in a moderate expression of enzyme in the CNS, reduction of GM1 storage, significant retention of motor function and a significant increase in lifespan. Interestingly, the therapeutic effect was more robust in females. Intracranial injections of AAVrh8 vector expressing high levels of βgal resulted in enzyme spread throughout the brain, significant retention of motor function and a significant increase in lifespan. Histological alterations were also found at the injection site in both βGal-/- and normal animals. We constructed a series of vectors with a range of decreasing enzyme expression levels to investigate the cause for the unanticipated result. Microarrays were performed on the injection site and we showed that a lower expressing AAVrh8-mβgal vector mitigated the negative response. Intracranial injection of this newly developed vector was shown to clear lysosomal storage throughout the CNS of βGal-/- mice. Taken together, these studies indicate that a combined systemic and fine-tuned intracranial approach may be the most effective in clearing lysosomal storage completely in the CNS while providing therapeutic benefit to the periphery.
Repository Citation
Weismann, CM. Approaches and Considerations Towards a Safe and Effective Adeno-Associated Virus Mediated Therapeutic Intervention for GM1-Gangliosidosis: A Dissertation. (2014). University of Massachusetts Medical School. GSBS Dissertations and Theses. Paper 767. DOI: 10.13028/M26K64. https://escholarship.umassmed.edu/gsbs_diss/767
DOI
10.13028/M26K64
DOI Link
Rights and Permissions
Copyright is held by the author, with all rights reserved.
Male animals near humane endpoint of untreated βGal-/- mice. βGal-/- + 3e11 vg at 258 days, βGal+/+ untreated at 258 days and βGal-/- + PBS at 221 days.
CMW - V2 F 3e11 260d.wmv (22605 kB)
Female animals near humane endpoint of untreated βGal-/- mice. βGal-/- + 3e11 vg at 260 days, βGal+/+ untreated at 260 days and βGal-/- + PBS at 236 days.
CMW - V3 M 1e11 263d.wmv (45746 kB)
Male animals near humane endpoint of untreated βGal-/- mice βGal-/- + 1e11 vg at 249 days, βGal+/- untreated at 264 days and βGal-/- + PBS at 263 days.
CMW - V4 F 1e11 264d.wmv (28754 kB)
Female animals near humane endpoint of untreated βGal-/- mice. βGal-/- + 3e11 vg at 258 days, βGal+/+ untreated at 278 days and βGal-/- + PBS at 264 days.
CMW - V5 F 3e11 590d.wmv (22865 kB)
Treated female βGal-/- + 3e11 vg at 590 days.
CMW - V6 F 3E11 566d.wmv (22999 kB)
Treated female βGal-/- + 3e11 vg at 563 days.
Included in
Genetics and Genomics Commons, Nervous System Diseases Commons, Neuroscience and Neurobiology Commons, Therapeutics Commons
Comments
This dissertation includes 6 videos that are referenced in Chapter II. See Additional Files available below.