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Full-Text Articles in Molecular Biology

Reprogramming The Retina: Next Generation Strategies Of Retinal Neuroprotection And Gene Therapy Vector Potency Assessment, Devin Scott Mcdougald Jan 2018

Reprogramming The Retina: Next Generation Strategies Of Retinal Neuroprotection And Gene Therapy Vector Potency Assessment, Devin Scott Mcdougald

Publicly Accessible Penn Dissertations

Mutations within over 250 known genes are associated with inherited retinal degeneration. Clinical success following gene replacement therapy for Leber’s congenital amaurosis type 2 establishes a platform for the development of downstream treatments targeting other forms of inherited and acquired ocular disease. Unfortunately, several challenges relevant to complex disease pathology and limitations of current gene transfer technologies impede the development of gene replacement for each specific form of retinal degeneration. Here we describe gene augmentation strategies mediated by recombinant AAV vectors that impede retinal degeneration in pre-clinical models of acquired and inherited vision loss. We demonstrate distinct neuroprotective effects ...


Gene Therapy Approaches To Immune Tolerance Induction In Canine Hemophilia, Robert French Jan 2018

Gene Therapy Approaches To Immune Tolerance Induction In Canine Hemophilia, Robert French

Publicly Accessible Penn Dissertations

A key issue in gene therapy is the immune response to the therapeutic transgene. This is especially important in applications where current treatments often elicit an antibody response, like hemophilia, where protein replacement therapy results in neutralizing

antibodies (“inhibitors”) in ~25% of severe hemophilia A and 1-3% of severe hemophilia B patients. To test the ability of skeletal muscle-directed gene therapy to prevent an immune response, we used an inhibitor-prone dog model of severe hemophilia B to express a hyperactive factor IX (FIX) variant from skeletal muscle via adeno-associated viral (AAV) vector and observed curative levels of expression that lasted ...


Pre-Clinical Development Of Aav Mediated Gene Therapy For Familial Lecithin Cholesterol Acyltransferase Deficiency, Devin Mae Christopher Jan 2017

Pre-Clinical Development Of Aav Mediated Gene Therapy For Familial Lecithin Cholesterol Acyltransferase Deficiency, Devin Mae Christopher

Publicly Accessible Penn Dissertations

Familial lecithin cholesterol acyltransferase (LCAT) deficiency (FLD), is a rare monogenic disease, characterized by abnormal lipid profile, corneal opacities, anemia and renal disease which progresses to renal failure. There is currently no approved treatment for patients with this disease. FLD is a good candidate for AAV mediated gene therapy but this therapeutic avenue has not been pursued until now. Here, we completed dose response studies in LCAT KO and LCAT KO/ human ApoA-I transgenic mice using AAV8 expressing human LCAT. AAV8-TBG-hLCAT induces significant increases in plasma % cholesterol esterified at a dose as low as 3e9 GC per mouse, which is ...


Intrathecal Aav9.Trastuzumab Tumor Prophylaxis And Treatment In A Murine Xenograft Model Of Her2+ Breast Cancer Brain Metastases, William Thomas Rothwell Jan 2017

Intrathecal Aav9.Trastuzumab Tumor Prophylaxis And Treatment In A Murine Xenograft Model Of Her2+ Breast Cancer Brain Metastases, William Thomas Rothwell

Publicly Accessible Penn Dissertations

Breast cancer brain metastases (BCBM) are a deadly sequela of breast tumors that overexpress human epidermal growth factor receptor 2 (HER2). HER2+ BCBM affects approximately 17,000 women in the US every year. Median survival is 10-13 months from the time of diagnosis of central nervous system (CNS) disease. Current therapeutic interventions are invasive, toxic, and largely ineffective, leaving a clear, unmet need for targeted HER2+ BCBM treatments. Trastuzumab(Herceptin®) is a monoclonal antibody used to treat HER2+ breast cancer successfully, but systemic trastuzumab cannot bypass the blood-brain barrier (BBB). To solve this problem, we have developed an adeno-associated virus ...


Adeno-Associated Viral Vector-Driven Expression Of Coagulation Proteins For Treatment Of Hemophilias And Cancer, Julie Marie Crudele Jan 2015

Adeno-Associated Viral Vector-Driven Expression Of Coagulation Proteins For Treatment Of Hemophilias And Cancer, Julie Marie Crudele

Publicly Accessible Penn Dissertations

Treatment of hemophilia, which involves infusion of the missing clotting factor, is often hindered by the development of neutralizing antibodies to the replaced clotting factor. We utilized liver-directed AAV gene therapy to tolerize outbred hemophiliac dogs with pre-existing anti-factor VIII and IX antibodies and to treat their underlying hemophilia. Additionally, we sought to shed light on the immunologic mechanisms responsible for this tolerization. Staining for CD4+CD25+FoxP3+ T cells and cytokine profiles of treated dogs suggest that induced Tregs are at least partially responsible for inducing and maintaining tolerance.

The second part of the dissertation attempts to determine the ...


Intrathecal Adeno-Associated Virus Vector Delivery For Mucopolysaccharidosis Type I, Christian Hinderer Jan 2015

Intrathecal Adeno-Associated Virus Vector Delivery For Mucopolysaccharidosis Type I, Christian Hinderer

Publicly Accessible Penn Dissertations

Mucopolysaccharidosis type I (MPS I) is a lysosomal storage disease resulting from deficiency of the enzyme α-L-iduronidase (IDUA). Accumulation of the IDUA substrates heparan and dermatan sulfate causes widespread organ pathology. While many of the somatic manifestations of MPS I can be treated with intravenous enzyme replacement, the devastating CNS sequelae—cognitive impairment, spinal cord compression, and hydrocephalus—do not respond to treatment. Partial preservation of cognitive function is possible with early hematopoietic stem cell transplantation, although transplant is associated with substantial morbidity and mortality. Gene transfer using adeno-associated virus (AAV) vectors offers a potential alternative approach to deliver the ...