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In vitro stem cell modelling demonstrates a proof-of-concept for excess functional mutant TIMP3 as the cause of Sorsby Fundus Dystrophy

In vitro stem cell modelling demonstrates a proof-of-concept for excess functional mutant TIMP3 as the cause of Sorsby Fundus Dystrophy
In vitro stem cell modelling demonstrates a proof-of-concept for excess functional mutant TIMP3 as the cause of Sorsby Fundus Dystrophy
Sorsby Fundus Dystrophy (SFD) is a rare autosomal dominant disease of the macula that leads to bilateral loss of central vision and is caused by mutations in the TIMP3 gene. However, the mechanisms by which TIMP3 mutations cause SFD are poorly understood. Here, we generated human induced pluripotent stem cell-derived retinal pigmented epithelial (hiPSC-RPE) cells from three SFD patients carrying TIMP3 p.(Ser204Cys) and three non-affected controls to study disease related structural and functional differences in the RPE. SFD-hiPSC-RPE exhibited characteristic RPE structure and physiology but showed significantly reduced transepithelial electrical resistance associated with enriched expression of cytoskeletal remodelling proteins. SFD-hiPSC-RPE exhibited basolateral accumulation of TIMP3 monomers, despite no change in TIMP3 gene expression. TIMP3 dimers were observed in both SFD and control hiPSC-RPE, suggesting mutant TIMP3 dimerization does not drive SFD pathology. Furthermore, mutant TIMP3 retained matrix metalloproteinase activity. Proteomic profiling showed increased expression of extracellular matrix proteins, endothelial cell interactions and angiogenesis-related pathways in SFD-hiPSC-RPE. By contrast, there were no changes in VEGF secretion. However, SFD-iPSC-RPE secreted higher levels of monocyte chemoattractant protein 1, platelet-derived growth factor, and angiogenin. Our findings provide a proof-of-concept that SFD patient-derived hiPSC-RPE mimic mature RPE cells and support the hypothesis that excess accumulation of mutant TIMP3, rather than an absence or deficiency of functional TIMP3, drives ECM and angiogenesis related changes in SFD.
Sorsby fundus dystrophy, human induced pluripotent stem cell, metalloproteinase inhibitor 3, retinal degeneration, retinal pigment epithelial cell
1096-9896
138-150
Dewing, Jennifer
4c4af0cf-b622-4f1c-aced-51a844e60475
Christensen, David RG
83abefba-97f2-4aea-b2fd-9516e3d2d99b
Scott, Jennifer
bdc803de-3082-4727-a4ca-f5a1cf3fcfcc
Cree, Angela
6724b71b-8828-4abb-971f-0856c2af555e
Ratnayaka, J. Arjuna
002499b8-1a9f-45b6-9539-5ac145799dfd
Lotery, Andrew
5ecc2d2d-d0b4-468f-ad2c-df7156f8e514
Dewing, Jennifer
4c4af0cf-b622-4f1c-aced-51a844e60475
Christensen, David RG
83abefba-97f2-4aea-b2fd-9516e3d2d99b
Scott, Jennifer
bdc803de-3082-4727-a4ca-f5a1cf3fcfcc
Cree, Angela
6724b71b-8828-4abb-971f-0856c2af555e
Ratnayaka, J. Arjuna
002499b8-1a9f-45b6-9539-5ac145799dfd
Lotery, Andrew
5ecc2d2d-d0b4-468f-ad2c-df7156f8e514

Dewing, Jennifer, Christensen, David RG, Scott, Jennifer, Cree, Angela, Ratnayaka, J. Arjuna and Lotery, Andrew (2020) In vitro stem cell modelling demonstrates a proof-of-concept for excess functional mutant TIMP3 as the cause of Sorsby Fundus Dystrophy. The Journal of Pathology, 252 (2), 138-150. (doi:10.1002/path.5506).

Record type: Article

Abstract

Sorsby Fundus Dystrophy (SFD) is a rare autosomal dominant disease of the macula that leads to bilateral loss of central vision and is caused by mutations in the TIMP3 gene. However, the mechanisms by which TIMP3 mutations cause SFD are poorly understood. Here, we generated human induced pluripotent stem cell-derived retinal pigmented epithelial (hiPSC-RPE) cells from three SFD patients carrying TIMP3 p.(Ser204Cys) and three non-affected controls to study disease related structural and functional differences in the RPE. SFD-hiPSC-RPE exhibited characteristic RPE structure and physiology but showed significantly reduced transepithelial electrical resistance associated with enriched expression of cytoskeletal remodelling proteins. SFD-hiPSC-RPE exhibited basolateral accumulation of TIMP3 monomers, despite no change in TIMP3 gene expression. TIMP3 dimers were observed in both SFD and control hiPSC-RPE, suggesting mutant TIMP3 dimerization does not drive SFD pathology. Furthermore, mutant TIMP3 retained matrix metalloproteinase activity. Proteomic profiling showed increased expression of extracellular matrix proteins, endothelial cell interactions and angiogenesis-related pathways in SFD-hiPSC-RPE. By contrast, there were no changes in VEGF secretion. However, SFD-iPSC-RPE secreted higher levels of monocyte chemoattractant protein 1, platelet-derived growth factor, and angiogenin. Our findings provide a proof-of-concept that SFD patient-derived hiPSC-RPE mimic mature RPE cells and support the hypothesis that excess accumulation of mutant TIMP3, rather than an absence or deficiency of functional TIMP3, drives ECM and angiogenesis related changes in SFD.

Text
Accepted version. In-vitro stem cell modelling demonstrates a proof-of-concept for excess functional mutant TIMP3 as the cause of Sorsby Fundus Dystrophy - Accepted Manuscript
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Accepted/In Press date: 29 June 2020
e-pub ahead of print date: 15 July 2020
Published date: 1 October 2020
Additional Information: © 2020 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Keywords: Sorsby fundus dystrophy, human induced pluripotent stem cell, metalloproteinase inhibitor 3, retinal degeneration, retinal pigment epithelial cell

Identifiers

Local EPrints ID: 442782
URI: http://eprints.soton.ac.uk/id/eprint/442782
ISSN: 1096-9896
PURE UUID: f3a7b7d6-0475-4ceb-add6-04c7fc497c73
ORCID for J. Arjuna Ratnayaka: ORCID iD orcid.org/0000-0002-1027-6938
ORCID for Andrew Lotery: ORCID iD orcid.org/0000-0001-5541-4305

Catalogue record

Date deposited: 27 Jul 2020 16:30
Last modified: 26 Nov 2021 07:18

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Contributors

Author: Jennifer Dewing
Author: David RG Christensen
Author: Jennifer Scott
Author: Angela Cree
Author: Andrew Lotery ORCID iD

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