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Development of a gene therapy for muscular dystrophy and the broader implications of immune responses to vector encoded antigens / Leon Morales.
- Format:
- Book
- Thesis/Dissertation
- Author/Creator:
- Morales, Juan Carlos, -2010, author.
- Language:
- English
- Subjects (All):
- Translation studies.
- Molecular biology.
- COVID-19.
- Cell and molecular biology--Penn dissertations.
- Penn dissertations--Cell and molecular biology.
- Local Subjects:
- Translation studies.
- Molecular biology.
- COVID-19.
- Cell and molecular biology--Penn dissertations.
- Penn dissertations--Cell and molecular biology.
- Genre:
- Academic theses.
- Physical Description:
- 1 online resource (102 pages)
- Contained In:
- Dissertations Abstracts International 82-07B.
- Place of Publication:
- [Philadelphia, Pennsylvania] : University of Pennsylvania ; Ann Arbor : ProQuest Dissertations & Theses, 2020.
- Language Note:
- English
- System Details:
- Mode of access: World Wide Web.
- text file
- Summary:
- Duchenne Muscular Dystrophy (DMD) is a devastating progressive muscle wasting disorder caused by mutations on the DMD gene that result in the complete absence of its protein product, dystrophin. Studies involving Adeno-associated viral (AAV) vector-mediated gene transfer approaches highlight the challenges the immune system can impose when it encounters "non-self" dystrophin epitopes. As an alternative, our lab relies on utrophin, a dystrophin paralog that retains most of dystrophin's structural and binding elements. Importantly, normal thymic expression in DMD subjects should protect utrophin by central immunologic tolerance. Here I describe how an AAV vector containing a codon optimized truncated utrophin transgene (AAV-μUtrophin) is a highly functional, non-immunogenic substitute for dystrophin, preventing the most deleterious histological and physiological aspects of muscular dystrophy in small and large animal models of DMD. Following systemic administration of AAV-μUtrophin in neonatal dystrophin-deficient mdx mice, histological and biochemical markers of myonecrosis and regeneration are completely suppressed throughout growth to adult weight. In the dystrophin-deficient golden retriever model, μUtrophin non-toxically prevented myonecrosis, even in the most powerful muscles. In a stringent test of immunogenicity, focal expression of μUtrophin in the deletional-null German shorthaired pointer canine model produced no evidence of cell-mediated immunity, in contrast to the robust T cell response against similarly constructed μDystrophin. These findings support a model in which utrophin-derived therapies might be used to treat clinical dystrophin deficiency, with a favorable immunologic profile. Our latter immunological findings would become the biological premise for a prospective vaccine against SARS-CoV-2 virus, the agent responsible for the ongoing global COVID-19 pandemic. SARS-CoV-2 relies on its major surface antigen, Spike protein, to gain access into host cells and therefore is a key target for vaccines. A single intramuscular injection in adult C57Bl/6 mice of a synthetic Spike protein (M8B), driven by a CMV promoter and packaged in an AAV vector (AAV-CMV-M8B), resulted in transient expression and a robust cell mediated immune response against M8B. Together these findings demonstrate how an AAV-mediated gene transfer approach can be used to provide a therapeutic effect and can be tuned to drive or steer away an immune response against vector encoded antigens.
- Notes:
- Source: Dissertations Abstracts International, Volume: 82-07, Section: B.
- Advisors: Stedman, Hansell H.; Bennett, Jean; Committee members: Nicola Mason; Tejvir Khurana; Michael Ostap; Valder Arruda.
- Department: Cell and Molecular Biology.
- Ph.D. University of Pennsylvania 2020.
- Local Notes:
- School code: 0175
- ISBN:
- 9798557052269
- Access Restriction:
- Restricted for use by site license.
- This item is not available from ProQuest Dissertations & Theses.
- This item must not be sold to any third party vendors.
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