Title:
Engineering Bovine Serum Albumin Nanoparticles For Improved Endosomal Escape And The Treatment Of Endometriosis

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WIMBERLEY-THESIS-2022.pdf (1.84 MB)
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Wimberley, Sydney C.
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Champion, Julie A.
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School of Chemical and Biomolecular Engineering
School established in 1901 as the School of Chemical Engineering; in 2003, renamed School of Chemical and Biomolecular Engineering
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https://hdl.handle.net/1853/73042
Abstract
Endometriosis is an estrogen driven condition that affects about 10% of menstruating women. It causes severe pain and infertility, and limited treatment options are available. This work uses the anti-inflammatory protein, AvrA, an effector protein found in Salmonella that inhibits nuclear factor-kB (NF-kB) and mitogen-activated protein kinase (MAPK) signal cascades. However, this protein is highly insoluble and requires a carrier for delivery to the cytosol, and bovine serum albumin (BSA) nanoparticles are used to deliver AvrA. AvrA-BSA nanoparticles were delivered to End1/E6E7, an epithelial cell line derived from a woman’s endometrium with endometriosis. To measure functionality of AvrA-BSA nanoparticles, inflammatory cytokines were measured in this cell type under inflammatory conditions. AvrA-BSA nanoparticles are internalized by cells using endocytosis. Their delivery to the cytosol is highly inefficient and endosome contents are trapped and later destroyed or recycled out of the cell following fusion with lysosomes, this is phenomenon is called endosomal entrapment. To overcome this entrapment, BSA nanoparticles were modified by conjugating histidine to their hydroxyl groups. Histidine’s variable chain is imidazole, and it can act as a buffer in lower pH environments, such as endosomes. In an endosome at lower pH protons and ions will enter the endosome causing them to swell due to a concentration gradient and an increase in osmotic pressure, eventually causes rupture. Endosomal escape of nanoparticles was evaluated using a Galectin-8 assay, to quantify endosomal disruption events; and a functional readout, where nanoparticles are loaded with toxic proteins allowing cell death to be an indication of nanoparticle escape. Overall, imidazole conjugated BSA nanoparticles do increase endosomal disruption events, and are able to incur an increased cell death when nanoparticles are loaded with a toxic protein.
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2022-12-09
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