Graduation Date

12-2018

Document Type

Master's Thesis

Degree Name

Master of Science

Department or Program

Biological Sciences

Department or Program Chair

Meredith Protas, PhD

First Reader

Manuel Lopez, PhD

Second Reader

Kristylea Ojeda, PhD

Abstract

Cyclodextrin (CD) is a cyclic oligosaccharide that has common use clinically as an excipient for drug stability and enhanced delivery. Currently, a highly soluble CD, hydroxypropyl-β-cyclodextrin (HP-bCD), is in phase 2b/3 clinical trials to treat patients with Niemann-Pick Type C disease (NPC). NPC is a lysosomal storage disorder where cholesterol is depleted from the cell membrane, yet it accumulates in the lysosomes/late endosomes along with other lipids and sterols. We have engineered a variety of functionalized versions of bCD to trace its function within the cell and improve upon its therapeutic performance. The modified bCDs have aided in demonstrating bCD cellular internalization and localization to the NPC1 late endosomal/lysosomal compartment. Filipin staining for free cholesterol accumulation and bCD mass spectrometry analysis suggest that the rate of intracellular uptake of bCD is limited by the concentration of bCD on the exterior of the cell. The amount of storage reduction seen in NPC cells is in turn dependent on the internal cellular concentration of bCD. Thus, bCDs designed for enhanced membrane or receptor mediated uptake were more potent at reducing cholesterol storage in a NPC1 null cell line.

Available for download on Thursday, November 25, 2021

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