Browsing by Subject "Epidermolysis Bullosa"

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    Charged Glycopolymer Materials for Epidermolysis Bullosa
    (2017-12) Boyle, William
    Improved delivery of therapeutic nucleic acid payloads to cells could lead to dramatically improved clinical outcomes for patients suffering from genetic disorders. This work focuses on the use of a trehalose-containing cationic glycopolymer, termed Tr4, to transfect clinically relevant cell types. In particular, the development of gene delivery methods to improve the transfection of cell types associated with the skin disease epidermolysis bullosa are investigated. The sulfated glycosaminoglycan, heparin, is show to form ternary complexes with pDNA and Tr4 leading to dramatically increased transfection efficiency in primary fibroblasts, induced pluripotent stem cells, HepG2, and U87-MG cells. This increase is not caused by improved uptake, but instead appears to be driven by improved intracellular trafficking of polyplexes compared transfection without heparin. Increasing the size of the plasmid cargo from 4.7 kbp to a more therapeutically relevant 10 kbp leads to the complete loss of transfection efficiency in Tr4-heparin transfection of primary fibroblasts and a reduction in transfection efficiency in iPSCs. Co-transfecting with additives meant to increase nuclear localization of the pDNA recovers the efficiency lost by increasing the plasmid size. These techniques allowed for the development of function transfection methods in iPSCs delivering a synthetic transcription activator of collagen type VII. Finally, nanofiber mats containing chondroitin sulfate were developed to scavenge inflammatory molecules from wound exudate.
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    Regulation of Type VII Collagen in Patients with Recessive Dystrophic Epidermolysis Bullosa
    (2017-08) Vanden Oever, Michael
    Recessive Dystrophic Epidermolysis Bullosa (RDEB) is a complex, life-threatening genetic skin disorder with painful complications. Currently, there is no cure, but there have been recent developments in both the basic biological research aspects and the translational therapies which make effectively treating this disease more likely in the near future. These advances include the use of stem cells and gene editing as well as new insights into the molecular mechanisms for certain aspects of RDEB pathology. The overall goals of our lab are to better define and characterize RDEB pathology, develop novel approaches for treating RDEB, and to improve upon the ways in which we analyze and understand the outcomes of those therapies. These goals are inherently dependent upon a comprehensive understanding of how type VII collagen is regulated, both during the normal wound healing process and over the course of therapeutic intervention. To that aim, we set out to understand two aspects of regulation of type VII collagen that were poorly understood: one focused on the nature of type VII collagen regulation during wound healing and the other focused on type VII collagen regulation during the hematopoietic stem cell transplantation process. We identified a particular micro RNA, miR-29, which regulates COL7A1 transcriptionally and post-transcriptionally. We also demonstrated that fludarabine, a key component of the hematopoietic cell transplantation (HCT) preparative regimen, modulates type VII collagen expression during hematopoietic stem cell transplantation. Our studies have identified a novel mechanism of regulation for type VII collagen that will hopefully give valuable insight into how to treat RDEB, ameliorate RDEB pathology, and properly evaluate clinical outcomes in patients that have receive HCT to treat RDEB.