Using human induced pluripotent stem cells to model cystinosis
type: CreativeWork
description: Cystinosis is a rare lysosomal storage disease caused by mutations in the CYSTINOSIN (CTNS) gene, encoding a cystine transporter located on the lysosomal membrane. Cystinosis leads to an accumulation of cystine within lysosomes in most cells of the body, however the kidney is the main organ affected. For poorly understood reasons, renal proximal tubule (PT) cells lose their functionality and degenerate in cystinosis, forming a characteristic “swanneck” lesion that leads to kidney failure before 10 years of age. Although, a drug-based therapy is available for cystinotic patients, the progression to kidney failure can only be delayed not prevented. A major challenge in the cystinosis field is the variation in cellular phenotypes between cell-based models of cystinosis and differences in the presentation of the disease in the mouse knockout. As such, there is no consensus on the pathways responsible for the cause of the disease. In order to develop a better human cell-based model of cystinosis, our lab has generated human pluripotent stem cells from a cystinosis patient (CTNS-iPS). The aim of this thesis is to characterise the cellular phenotype of CTNS-iPS cells and to develop protocols to differentiate these cells into cystinotic PT cells for further study. CTNS-iPS cells were found to exhibit classic cystinotic defects including increased cystine levels, an altered ratio of reduced to oxidised glutathione (indicative of oxidative stress), mitochondria with a fragmented morphology, and enlarged late endosomes/lysosomes that cluster in a perinuclear region. Approximately 25% of CTNS-iPS cells display massive multivesicular body-like vesicles that contain numerous electron-lucent vesicles when visualised by electron microscopy. Two protocols were established to convert iPS cells into PT cells. In the first, iPS cells were treated with Activin A, the Wnt agonist CHIR99021 (CHIR) and BMPs and then purified by FACS, based on binding to the PT cell specific lectin FITC-con…
abstract: Cystinosis is a rare lysosomal storage disease caused by mutations in the CYSTINOSIN (CTNS) gene, encoding a cystine transporter located on the lysosomal membrane. Cystinosis leads to an accumulation of cystine within lysosomes in most cells of the body, however the kidney is the main organ affected. For poorly understood reasons, renal proximal tubule (PT) cells lose their functionality and degenerate in cystinosis, forming a characteristic “swanneck” lesion that leads to kidney failure before 10 years of age. Although, a drug-based therapy is available for cystinotic patients, the progression to kidney failure can only be delayed not prevented. A major challenge in the cystinosis field is the variation in cellular phenotypes between cell-based models of cystinosis and differences in the presentation of the disease in the mouse knockout. As such, there is no consensus on the pathways responsible for the cause of the disease. In order to develop a better human cell-based model of cystinosis, our lab has generated human pluripotent stem cells from a cystinosis patient (CTNS-iPS). The aim of this thesis is to characterise the cellular phenotype of CTNS-iPS cells and to develop protocols to differentiate these cells into cystinotic PT cells for further study. CTNS-iPS cells were found to exhibit classic cystinotic defects including increased cystine levels, an altered ratio of reduced to oxidised glutathione (indicative of oxidative stress), mitochondria with a fragmented morphology, and enlarged late endosomes/lysosomes that cluster in a perinuclear region. Approximately 25% of CTNS-iPS cells display massive multivesicular body-like vesicles that contain numerous electron-lucent vesicles when visualised by electron microscopy. Two protocols were established to convert iPS cells into PT cells. In the first, iPS cells were treated with Activin A, the Wnt agonist CHIR99021 (CHIR) and BMPs and then purified by FACS, based on binding to the PT cell specific lectin FITC-con…
author: Przepiorski, Aneta
license: https://researchspace.auckland.ac.nz/docs/uoa-docs/rights.htm
date published: 2017
date modified: 2026-07-24T01:02:59Z
provider: University of Auckland
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: ResearchSpace@Auckland

PropertyValue
property ID: Handle
value: https://hdl.handle.net/2292/33556