Journal article
Lentiviral mediated delivery of CRISPR/Cas9 reduces intraocular pressure in a mouse model of myocilin glaucoma
Scientific reports, Vol.14(1), 6958
03/23/2024
DOI: 10.1038/s41598-024-57286-6
PMCID: PMC10960846
PMID: 38521856
Abstract
Mutations in myocilin (MYOC) are the leading known genetic cause of primary open-angle glaucoma, responsible for about 4% of all cases. Mutations in MYOC cause a gain-of-function phenotype in which mutant myocilin accumulates in the endoplasmic reticulum (ER) leading to ER stress and trabecular meshwork (TM) cell death. Therefore, knocking out myocilin at the genome level is an ideal strategy to permanently cure the disease. We have previously utilized CRISPR/Cas9 genome editing successfully to target MYOC using adenovirus 5 (Ad5). However, Ad5 is not a suitable vector for clinical use. Here, we sought to determine the efficacy of adeno-associated viruses (AAVs) and lentiviruses (LVs) to target the TM. First, we examined the TM tropism of single-stranded (ss) and self-complimentary (sc) AAV serotypes as well as LV expressing GFP via intravitreal (IVT) and intracameral (IC) injections. We observed that LV_GFP expression was more specific to the TM injected via the IVT route. IC injections of Trp-mutant scAAV2 showed a prominent expression of GFP in the TM. However, robust GFP expression was also observed in the ciliary body and retina. We next constructed lentiviral particles expressing Cas9 and guide RNA (gRNA) targeting MYOC (crMYOC) and transduction of TM cells stably expressing mutant myocilin with LV_crMYOC significantly reduced myocilin accumulation and its associated chronic ER stress. A single IVT injection of LV_crMYOC in Tg-MYOC
mice decreased myocilin accumulation in TM and reduced elevated IOP significantly. Together, our data indicates, LV_crMYOC targets MYOC gene editing in TM and rescues a mouse model of myocilin-associated glaucoma.
Details
- Title: Subtitle
- Lentiviral mediated delivery of CRISPR/Cas9 reduces intraocular pressure in a mouse model of myocilin glaucoma
- Creators
- Shruti V Patil - University of North Texas Health Science CenterBalasankara Reddy Kaipa - University of California, IrvineSujata Ranshing - University of North Texas Health Science CenterYogapriya Sundaresan - University of California, IrvineJ Cameron Millar - University of North Texas Health Science CenterBhavani Nagarajan - University of North Texas Health Science CenterCharles Kiehlbauch - University of North Texas Health Science CenterQihong Zhang - University of IowaAnkur Jain - University of IowaCharles C Searby - University of IowaTodd E Scheetz - University of IowaAbbot F Clark - University of North Texas Health Science CenterVal C Sheffield - University of IowaGulab S Zode - University of California, Irvine
- Resource Type
- Journal article
- Publication Details
- Scientific reports, Vol.14(1), 6958
- DOI
- 10.1038/s41598-024-57286-6
- PMID
- 38521856
- PMCID
- PMC10960846
- eISSN
- 2045-2322
- Grant note
- EY034070 / NEI NIH HHS EY026177 / NEI NIH HHS EY030366 / NEI NIH HHS EY025580 / NEI NIH HHS
- Language
- English
- Date published
- 03/23/2024
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; Stead Family Department of Pediatrics; Iowa Neuroscience Institute; Medical Genetics and Genomics; Ophthalmology and Visual Sciences
- Record Identifier
- 9984577112802771
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