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Background
Vanishing white matter disease (VWMD) is a rare leukodystrophy caused by mutations of the guanine exchange factor eIF2B that typically presents with juvenile onset. There are few treatments and no cures for the disease. Recent progress in the field has established mitochondrial dysfunction and endoplasmic reticulum (ER) stress to be strongly implicated in observed glial cell pathology. Drug repurposing offers a rapid approach toward translation of therapeutics with already-licensed drugs.
Objective
The aim of this study was to use fibroblasts and induced pluripotent stem cell (iPSC)-derived astrocytes from patients bearing the EIF2B5R113H/A403V or EIF2B2G200V/E213G VWMD mutations to identify potentially repurposable FDA-approved drugs based on in vitro assays.
Methods
Cell viability in the presence or absence of stress was assessed by resazurin reduction activity assay, mitochondrial membrane potential by TMRE fluorescence, and oxidative stress by H2DCFDA oxidation. Relative eIF2B phosphorylation, GADD34 and CHOP were quantified by fluorescent western blot.
Results
Dysregulated GADD34 and CHOP were identified in patient fibroblasts and iPSC-derived astrocytes under induced stress conditions. A drug screen from a 2,400 FDA-approved drug library with EIF2B5R113H/A403V VWMD patient fibroblasts identified 113 anti-inflammatory drugs as a major class of hits with cytoprotective effects. A panel of potential candidate drugs, including berberine, deflazacort, ursodiol, zileuton, guanabenz and Anavex 2–73, and preclinical ISRIB, increased cell survival of EIF2B5R113H/A403V or EIF2B2G200V/E213G VWMD astrocytes, and were further investigated for their effect on the integrated stress response and mitochondrial stress. Ursodiol demonstrated capacity to ameliorate oxidative stress and loss of mitochondrial membrane potential in VWMD patient iPSC-derived astrocytes in the presence or absence of stress conditions.
Conclusion
Patient-derived cells can be used to identify cellular phenotypes and for large-scale drug screening. Anti-inflammatory compounds, such as berberine, deflazacort, ursodiol and zileuton are potentially repurposable drug candidates for VWMD that should be further investigated for translation in vivo.
Keywords
Vanishing white matter disease, leukodystrophy, cell stress, drug repurposing, induced pluripotent stem cells, astrocytes
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CURRENT STATUS: Published
View the published article at Translational Medicine Communications.
No community comments so far
Review #1 received
Received 06 Oct, 2020
Editorial decision: Accept
On 06 Oct, 2020
Reviewer #2 agreed
On 05 Oct, 2020
Review #2 received
Received 05 Oct, 2020
Reviewer #1 agreed
On 30 Sep, 2020
Editor assigned
On 28 Sep, 2020
Reviewers invited
Invitations sent on 28 Sep, 2020
Submission checks complete
On 27 Sep, 2020
Editor invited
On 27 Sep, 2020
Version 1
Posted 26 Aug, 2020
No comments provided
Editorial decision: Minor revision
On 14 Sep, 2020
Review #2 received
Received 12 Sep, 2020
Review #1 received
Received 06 Sep, 2020
Reviewer #2 agreed
On 29 Aug, 2020
Reviewer #1 agreed
On 26 Aug, 2020
Reviewers invited
Invitations sent on 25 Aug, 2020
Editor assigned
On 19 Aug, 2020
First submitted
On 18 Aug, 2020
Submission checks complete
On 18 Aug, 2020
Editor invited
On 18 Aug, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Translational Medicine
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See the published version of this preprint at Translational Medicine Communications.
This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Published
View the published article at Translational Medicine Communications.
No community comments so far
Review #1 received
Received 06 Oct, 2020
Editorial decision: Accept
On 06 Oct, 2020
Reviewer #2 agreed
On 05 Oct, 2020
Review #2 received
Received 05 Oct, 2020
Reviewer #1 agreed
On 30 Sep, 2020
Editor assigned
On 28 Sep, 2020
Reviewers invited
Invitations sent on 28 Sep, 2020
Submission checks complete
On 27 Sep, 2020
Editor invited
On 27 Sep, 2020
Version 1
Posted 26 Aug, 2020
No comments provided
Editorial decision: Minor revision
On 14 Sep, 2020
Review #2 received
Received 12 Sep, 2020
Review #1 received
Received 06 Sep, 2020
Reviewer #2 agreed
On 29 Aug, 2020
Reviewer #1 agreed
On 26 Aug, 2020
Reviewers invited
Invitations sent on 25 Aug, 2020
Editor assigned
On 19 Aug, 2020
First submitted
On 18 Aug, 2020
Submission checks complete
On 18 Aug, 2020
Editor invited
On 18 Aug, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Translational Medicine
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published article at Translational Medicine Communications.
Background
Vanishing white matter disease (VWMD) is a rare leukodystrophy caused by mutations of the guanine exchange factor eIF2B that typically presents with juvenile onset. There are few treatments and no cures for the disease. Recent progress in the field has established mitochondrial dysfunction and endoplasmic reticulum (ER) stress to be strongly implicated in observed glial cell pathology. Drug repurposing offers a rapid approach toward translation of therapeutics with already-licensed drugs.
Objective
The aim of this study was to use fibroblasts and induced pluripotent stem cell (iPSC)-derived astrocytes from patients bearing the EIF2B5R113H/A403V or EIF2B2G200V/E213G VWMD mutations to identify potentially repurposable FDA-approved drugs based on in vitro assays.
Methods
Cell viability in the presence or absence of stress was assessed by resazurin reduction activity assay, mitochondrial membrane potential by TMRE fluorescence, and oxidative stress by H2DCFDA oxidation. Relative eIF2B phosphorylation, GADD34 and CHOP were quantified by fluorescent western blot.
Results
Dysregulated GADD34 and CHOP were identified in patient fibroblasts and iPSC-derived astrocytes under induced stress conditions. A drug screen from a 2,400 FDA-approved drug library with EIF2B5R113H/A403V VWMD patient fibroblasts identified 113 anti-inflammatory drugs as a major class of hits with cytoprotective effects. A panel of potential candidate drugs, including berberine, deflazacort, ursodiol, zileuton, guanabenz and Anavex 2–73, and preclinical ISRIB, increased cell survival of EIF2B5R113H/A403V or EIF2B2G200V/E213G VWMD astrocytes, and were further investigated for their effect on the integrated stress response and mitochondrial stress. Ursodiol demonstrated capacity to ameliorate oxidative stress and loss of mitochondrial membrane potential in VWMD patient iPSC-derived astrocytes in the presence or absence of stress conditions.
Conclusion
Patient-derived cells can be used to identify cellular phenotypes and for large-scale drug screening. Anti-inflammatory compounds, such as berberine, deflazacort, ursodiol and zileuton are potentially repurposable drug candidates for VWMD that should be further investigated for translation in vivo.
Figure 1
Figure 2
Figure 3
Figure 4
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