Mouse γ-Synuclein Promoter-Mediated Gene Expression and Editing in Mammalian Retinal Ganglion Cells

Qizhao Wang; Pei Zhuang; Haoliang Huang; Liang Li; Liang Liu; Hannah C. Webber; Roopa Dalal; Leonard Siew; Clarisse M. Fligor; Kun-Che Chang; Michael Nahmou; Alexander Kreymerman; Jason S. Meyer; Jeffrey Louis Goldberg; Yang Hu

doi:10.1523/jneurosci.0102-20.2020

journal article Open Access Apr 16, 2020

Mouse γ-Synuclein Promoter-Mediated Gene Expression and Editing in Mammalian Retinal Ganglion Cells

Qizhao Wang

Pei Zhuang Haoliang Huang

Liang Li

Liang Liu

Hannah C. Webber Roopa Dalal Leonard Siew Clarisse M. Fligor Kun-Che Chang

Michael Nahmou Alexander Kreymerman Jason S. Meyer Jeffrey Louis Goldberg Yang Hu

The Journal of Neuroscience Vol. 40 No. 20 pp. 3896-3914 · Society for Neuroscience

View at Publisher Save 10.1523/jneurosci.0102-20.2020

Abstract

Optic neuropathies are a group of optic nerve (ON) diseases caused by various insults including glaucoma, inflammation, ischemia, trauma, and genetic deficits, which are characterized by retinal ganglion cell (RGC) death and ON degeneration. An increasing number of genes involved in RGC intrinsic signaling have been found to be promising neural repair targets that can potentially be modulated directly by gene therapy, if we can achieve RGC specific gene targeting. To address this challenge, we first used adeno-associated virus (AAV)-mediated gene transfer to perform a low-throughputin vivoscreening in both male and female mouse eyes and identified the mouse γ-synuclein (mSncg) promoter, which specifically and potently sustained transgene expression in mouse RGCs and also works in human RGCs. We further demonstrated that gene therapy that combines AAV-mSncg promoter with clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 gene editing can knock down pro-degenerative genes in RGCs and provide effective neuroprotection in optic neuropathies.SIGNIFICANCE STATEMENTHere, we present an RGC-specific promoter, mouse γ-synuclein (mSncg) promoter, and perform extensive characterization and proof-of-concept studies of mSncg promoter-mediated gene expression and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 gene editing in RGCsin vivo. To our knowledge, this is the first report demonstratingin vivoneuroprotection of injured RGCs and optic nerve (ON) by AAV-mediated CRISPR/Cas9 inhibition of genes that are critical for neurodegeneration. It represents a powerful tool to achieve RGC-specific gene modulation, and also opens up a promising gene therapy strategy for optic neuropathies, the most common form of eye diseases that cause irreversible blindness.

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Details

Published: Apr 16, 2020
Vol/Issue: 40(20)
Pages: 3896-3914
License: View

Authors

Department of Ophthalmology, Stanford University School of Medicine, Palo Alto, United States

H

Haoliang Huang

Department of Ophthalmology, Stanford University School of Medicine, Palo Alto, United States

Fudan University

Department of Ophthalmology, Stanford University School of Medicine, Palo Alto, United States

Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Science, University of Colorado Anschutz Medical Campus, Aurora, Colorado 80045, United States; Department of Ophthalmology, School of Medicine, University of Colorado Anschutz Medical Campus, Aurora, Colorado 80045, United States

M

Michael Nahmou

Byers Eye Institute and Spencer Center for Vision Research, Department of Ophthalmology, Stanford University, Palo Alto, California, United States

Jeffrey Louis Goldberg

Y

Yang Hu

Comprehensive Cancer Center and Departments of Internal Medicine, Pharmacology, and Medicinal Chemistry, University of Michigan, Ann Arbor, Michigan 48109, United States

Cite This Article

Qizhao Wang, Pei Zhuang, Haoliang Huang, et al. (2020). Mouse γ-Synuclein Promoter-Mediated Gene Expression and Editing in Mammalian Retinal Ganglion Cells. The Journal of Neuroscience, 40(20), 3896-3914. https://doi.org/10.1523/jneurosci.0102-20.2020

Mouse γ-Synuclein Promoter-Mediated Gene Expression and Editing in Mammalian Retinal Ganglion Cells

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