Using a human induced pluripotent stem cell model to investigate the role of perlecan in cardiovascular development and disease.

Johnson, Benjamin Blakeley (2023) Using a human induced pluripotent stem cell model to investigate the role of perlecan in cardiovascular development and disease. Doctoral thesis, University of East Anglia.

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Abstract

Heparan sulfate proteoglycans (HSPGs) contribute to structural integrity and signalling regulation during cardiac development, healing, and wound repair, one such HSPG being perlecan (HSPG2). Perlecan is a large basement membrane protein known for the binding of growth factors and extracellular matrix, highlighted as essential in innate cardiac function. A HSPG2 knock-out (KO) causes embryonic lethality in mice due to ‘leaky’ hearts, this KO in rescued mice show perlecan has a role in metabolic output in a cell specific manner. During cardiomyocyte (CM) development, a metabolic switch from glycolysis to fatty acid metabolism is essential for developmental maturation. I aimed to investigate the role of perlecan in cardiac cell populations.

Perlecan deficient hiPSC-Cardiac fibroblasts (CFs) showed higher levels of myofibroblast activation, and a reduced mitochondrial function compared to WT. Transcriptomic analysis of a differentiation time course to hiPSC-CMs highlighted 11 genes consistently dysregulated due to perlecan deficiency, including HSPG2, NNAT, SLC15A4, KCNC3, RBM46, GLT1D1, TMSB4Y, ZNF676, SVIL-AS1, MGMT and TRIM61. Perlecan deficient hiPSC-CMs have an increased reliance of glycolysis, are prone to cardiac remodelling when in 3D and show increased genetic differences during the stages of CM maturation. Our results show how hiPSC-CMs with attenuated perlecan have an immature cardiac phenotype when compared to WT hiPSC-CMs, confirmed via the use of metabolic assays and gene expression. While culturing of hiPSC-CMs on a synthetic perlecan surface induced enhanced multinucleation, suggestive of hyperplastic to hypertrophic development.

The lack of perlecan in our cardiac populations has detrimental effects on metabolism of both hiPSC-CMs and CFs. The metabolic switch in hiPSC-CMs is essential to maturation, it is attenuated in our perlecan deficient hiPSC-CMs, indicative of a less mature CM. Understanding perlecans role in cardiac populations could lead to improved maturation of hiPSC-CMs for drug screening and disease modelling, as well as further insights into cardiac wound recovery.

Item Type: Thesis (Doctoral)
Faculty \ School: Faculty of Medicine and Health Sciences > Norwich Medical School
Depositing User: Chris White
Date Deposited: 21 Jun 2023 10:22
Last Modified: 29 Feb 2024 01:38
URI: https://ueaeprints.uea.ac.uk/id/eprint/92448
DOI:

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