CRISPR-Cas9–based treatment of myocilin-associated glaucoma
Edited by Donald J. Zack, Johns Hopkins University, Baltimore, MD, and accepted by Editorial Board Member Jeremy Nathans August 25, 2017 (received for review April 22, 2017)
Significance
A mutation in myocilin is the most common known genetic cause of primary open-angle glaucoma (POAG). These mutations, which are dominant in nature, affect trabecular meshwork (TM) health and/or function and cause elevated intraocular pressure. Using in vitro human trabecular meshwork cells, an in vivo mouse model, and ex vivo human eyes, our study demonstrates the potential of clustered regularly interspaced short palindromic repeats (CRISPR)-mediated genome editing in human myocilin-associated POAG. By disrupting the mutant myocilin gene and its function using CRISPR-Cas9, we were able to reduce associated endoplasmic reticulum stress, lower intraocular pressure, and prevent further glaucomatous damage in mouse eyes. We also show the feasibility of using the CRISPR-Cas9 system in cultured human eyes.
Abstract
Primary open-angle glaucoma (POAG) is a leading cause of irreversible vision loss worldwide, with elevated intraocular pressure (IOP) a major risk factor. Myocilin (MYOC) dominant gain-of-function mutations have been reported in ∼4% of POAG cases. MYOC mutations result in protein misfolding, leading to endoplasmic reticulum (ER) stress in the trabecular meshwork (TM), the tissue that regulates IOP. We use CRISPR-Cas9–mediated genome editing in cultured human TM cells and in a MYOC mouse model of POAG to knock down expression of mutant MYOC, resulting in relief of ER stress. In vivo genome editing results in lower IOP and prevents further glaucomatous damage. Importantly, using an ex vivo human organ culture system, we demonstrate the feasibility of human genome editing in the eye for this important disease.
Acknowledgments
This project was supported by National Institutes of Health Grants R01 EY024259, R01 EY026177, and R00 EY022077, funding from the Howard Hughes Medical Institute, and from the Roy J. Carver Charitable Trust. Adenoviral vectors were obtained from the Gene Transfer Vector Core at the University of Iowa.
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Freely available online through the PNAS open access option.
Submission history
Published online: October 2, 2017
Published in issue: October 17, 2017
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Acknowledgments
This project was supported by National Institutes of Health Grants R01 EY024259, R01 EY026177, and R00 EY022077, funding from the Howard Hughes Medical Institute, and from the Roy J. Carver Charitable Trust. Adenoviral vectors were obtained from the Gene Transfer Vector Core at the University of Iowa.
Notes
This article is a PNAS Direct Submission. D.J.Z. is a guest editor invited by the Editorial Board.
Authors
Competing Interests
The authors declare no conflict of interest.
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Cite this article
CRISPR-Cas9–based treatment of myocilin-associated glaucoma, Proc. Natl. Acad. Sci. U.S.A.
114 (42) 11199-11204,
https://doi.org/10.1073/pnas.1706193114
(2017).
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