Start codon disruption with CRISPR/Cas9 prevents murine Fuchs’ endothelial corneal dystrophy

AbstractA missense mutation of collagen type VIII alpha 2 chain (COL8A2) gene leads to early onset Fuchs’ endothelial corneal dystrophy (FECD), which progressively impairs vision through loss of corneal endothelial cells. We demonstrate that CRISPR/Cas9-based postnatal gene editing achieves structural and functional rescue in a mouse model of FECD. A single intraocular injection of an adenovirus encoding both the Cas9 gene and guide RNA (Ad-Cas9-Col8a2gRNA), efficiently knocked down mutant COL8A2 expression in corneal endothelial cells, prevented endothelial cell loss, and rescued corneal endothelium pumping function in adult Col8a2 mutant mice. There were no adverse sequelae on histology or electroretinography. Col8a2 start codon disruption represents a non-surgical strategy to prevent vision loss in early-onset FECD. As this demonstrates the ability of Ad-Cas9-gRNA to restore phenotype in adult post-mitotic cells, this method may be widely applicable to adult-onset diseases, even in tissues affected with disorders of non-reproducing cells.

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Start codon disruption with CRISPR/Cas9 prevents murine Fuchs' endothelial corneal dystrophy

eLife ◽

10.7554/elife.55637 ◽

2021 ◽

Vol 10 ◽

Author(s):

Hironori Uehara ◽

Xiaohui Zhang ◽

Felipe Pereira ◽

Siddharth Narendran ◽

Susie Choi ◽

...

Keyword(s):

Endothelial Cells ◽

Early Onset ◽

Vision Loss ◽

Cell Loss ◽

Corneal Dystrophy ◽

Start Codon ◽

Corneal Endothelial Cells ◽

Endothelial Cell Loss ◽

Guide Rna ◽

Pumping Function

A missense mutation of collagen type VIII alpha 2 chain (COL8A2) gene leads to early onset Fuchs' endothelial corneal dystrophy (FECD), which progressively impairs vision through loss of corneal endothelial cells. We demonstrate that CRISPR/Cas9-based postnatal gene editing achieves structural and functional rescue in a mouse model of FECD. A single intraocular injection of an adenovirus encoding both the Cas9 gene and guide RNA (Ad-Cas9-Col8a2gRNA), efficiently knocked down mutant COL8A2 expression in corneal endothelial cells, prevented endothelial cell loss, and rescued corneal endothelium pumping function in adult Col8a2 mutant mice. There were no adverse sequelae on histology or electroretinography. Col8a2 start codon disruption represents a non-surgical strategy to prevent vision loss in early-onset FECD. As this demonstrates the ability of Ad-Cas9-gRNA to restore phenotype in adult post-mitotic cells, this method may be widely applicable to adult-onset diseases, even in tissues affected with disorders of non-reproducing cells.

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Sodium Ferulate Attenuates Lidocaine-Induced Corneal Endothelial Impairment

Oxidative Medicine and Cellular Longevity ◽

10.1155/2018/4967318 ◽

2018 ◽

Vol 2018 ◽

pp. 1-8 ◽

Cited By ~ 1

Author(s):

Guojian Jiang ◽

Tingjun Fan

Keyword(s):

Endothelial Cells ◽

Endothelial Cell ◽

Morphological Changes ◽

Corneal Thickness ◽

Cell Loss ◽

Corneal Endothelial Cell ◽

Corneal Endothelial Cells ◽

Endothelial Cell Loss ◽

Sodium Ferulate ◽

Human Corneal Endothelial Cells

The introduction of intracameral anaesthesia by injection of lidocaine has become popular in cataract surgery for its inherent potency, rapid onset, tissue penetration, and efficiency. However, intracameral lidocaine causes corneal thickening, opacification, and corneal endothelial cell loss. Herein, we investigated the effects of lidocaine combined with sodium ferulate, an antioxidant with antiapoptotic and anti-inflammatory properties, on lidocaine-induced damage of corneal endothelia with in vitro experiment of morphological changes and cell viability of cultured human corneal endothelial cells and in vivo investigation of corneal endothelial cell density and central corneal thickness of cat eyes. Our finding indicates that sodium ferulate from 25 to 200 mg/L significantly reduced 2 g/L lidocaine-induced toxicity to human corneal endothelial cells, and 50 mg/L sodium ferulate recovered the damaged human corneal endothelial cells to normal growth status. Furthermore, 100 mg/L sodium ferulate significantly inhibited lidocaine-induced corneal endothelial cell loss and corneal thickening in cat eyes. In conclusion, sodium ferulate protects human corneal endothelial cells from lidocaine-induced cytotoxicity and attenuates corneal endothelial cell loss and central corneal thickening of cat eyes after intracameral injection with lidocaine. It is likely that the antioxidant effect of sodium ferulate reduces the cytotoxic and inflammatory corneal reaction during intracameral anaesthesia.

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Biomechanical changes to Descemet's membrane precede endothelial cell loss in an early-onset murine model of Fuchs endothelial corneal dystrophy

Experimental Eye Research ◽

10.1016/j.exer.2018.11.021 ◽

2019 ◽

Vol 180 ◽

pp. 18-22 ◽

Cited By ~ 3

Author(s):

Brian C. Leonard ◽

Iman Jalilian ◽

Vijay Krishna Raghunathan ◽

Wei Wang ◽

Albert S. Jun ◽

...

Keyword(s):

Endothelial Cell ◽

Murine Model ◽

Early Onset ◽

Cell Loss ◽

Corneal Dystrophy ◽

Endothelial Cell Loss ◽

Descemet's Membrane ◽

Descemet’S Membrane ◽

Biomechanical Changes

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Effect of Shear Stress on Attachment of Corneal Endothelial Cells in Association With Corneal Endothelial Cell Loss After Laser Iridotomy

Cornea ◽

10.1097/01.ico.0000178735.27674.52 ◽

2005 ◽

Vol 24 (Supplement 1) ◽

pp. S55-S58 ◽

Cited By ~ 20

Author(s):

Yuichi Kaji ◽

Tetsuro Oshika ◽

Tomohiko Usui ◽

Jun Sakakibara

Keyword(s):

Endothelial Cells ◽

Shear Stress ◽

Endothelial Cell ◽

Cell Loss ◽

Corneal Endothelial Cell ◽

Corneal Endothelial Cells ◽

Endothelial Cell Loss ◽

Laser Iridotomy

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Comparison of Cornea Endothelial Cells Density the After Aspiration of Dense Cataracts using Two Variants of Phacoemulsification Parameters

Ophthalmology in Russia ◽

10.18008/1816-5095-2018-2s-145-152 ◽

2018 ◽

Vol 15 (2S) ◽

pp. 145-152

Author(s):

S. V. Shukhaev ◽

E. V. Boiko

Keyword(s):

Endothelial Cells ◽

Postoperative Complications ◽

Cell Loss ◽

Main Group ◽

Cell Counting ◽

Control Group ◽

Corneal Endothelial Cells ◽

Significant Difference ◽

Ultrasound Energy ◽

And Control

Purpose: to compare two types of phacoemulsification parameters (combination ultrasound and torsional US with IP) with estimating the number of postoperative complications caused by intraoperative trauma of corneal endothelial cells.Patients and methods. 72 patients underwent phacoemulsification with IOL implantation. Patients were randomly divided into two groups (main n = 33 and control n = 39). During the aspiration of lens fragments in the main group the combination ultrasound was used, while torsional US with IP was used in the control group. Endothelial cell counting and other examinations were performed 1 day, 1 week and 6 months after the surgery.Results. CDVA in the explored groups 1 week after the surgery was similar: the main group — 0.813 ± 0.228, the control group — 0.765 ± 0.250, There was also no statistically significant difference in the thickness of the cornea between the groups: the main group — 533.48 ± 12.41, the control group — 536.44 ± 10.92. At the same time, a statistically significant difference was found in the density of endothelial cells: the main group: 1871.30 ± 187.41 (after 1 week), 1865 ± 178.9 (after 6 months); control group: 1809.63 ± 225.43 (after 1 week), 1791 ± 230.82 (after 6 months). The percentage of cell loss was, respectively, lower in the main group at all times of observation: 3.90% (after 1 day), 4.54% (after 1 week), 4.9% (after 6 months). In the control group: 7.71% (after 1 day), 9.25% (after 1 week), 10.4% (after 6 months).Conclusions. Data obtained in this study has showed the advantages of combination ultrasound in comparison with torsional US with IP, when performing aspiration of dense lenses. Due to lower consumption of ultrasound energy and higher aspiration rate of the fragments, combination ultrasound can reduce the loss of corneal endothelial cells and decrease the number and severity of postoperative complications associated with it.

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Potential functional restoration of corneal endothelial cells in Fuchs endothelial corneal dystrophy by ROCK inhibitor (ripasudil)

American Journal of Ophthalmology ◽

10.1016/j.ajo.2020.12.006 ◽

2020 ◽

Author(s):

Ursula Schlötzer-Schrehardt ◽

Matthias Zenkel ◽

Maria Strunz ◽

Andreas Gießl ◽

Hannah Schondorf ◽

...

Keyword(s):

Endothelial Cells ◽

Corneal Dystrophy ◽

Functional Restoration ◽

Corneal Endothelial Cells ◽

Rock Inhibitor

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A STUDY ON ENDOTHELIAL CELL LOSS AFTER OPTICAL KERATOPLASTY- A RETROSPECTIVE STUDY IN A TERTIARY EYE CARE CENTRE IN CHENNA

10.36106/9500847 ◽

2021 ◽

pp. 19-21

Author(s):

Rohini K ◽

Padmapriya R ◽

Niranjan Karthik Senthil Kumar

Keyword(s):

Endothelial Cells ◽

Retrospective Study ◽

Endothelial Cell ◽

Tertiary Care ◽

Cell Loss ◽

Care Centre ◽

Endothelial Cell Loss ◽

Tertiary Care Centre ◽

Triple Procedure ◽

Continuous Loss

Aim: a study of endothelial cell loss after optical keratoplasty in 50 patients in a tertiary care centre in Chennai. Materials and Methods: A retrospective study was carried out on 50 patients who underwent optical keratoplasty in a tertiary care centre in Chennai. 50 patients in the age group 11-80 were included in this study. Postoperatively they were periodically followed up regarding graft clarity, presence of vascularisation, vision and endothelial cell count. During follow up presence of any complications were assessed and recorded. At the end of 6 months, there was an average of 37.5% loss of endothelial ce Observation And Results: lls in PKP and 44.5% in LKPs and 38.6% in Triple procedure. At the end of one month, patients who had repeat Keratoplasty for failed graft had more loss (20.1%) whereas at the end of 6 months, Pseudophakic Bullous Keratopathy patients showed more loss (49.4%). These indicate migration of endothelial cells along a density gradient after keratoplasty. Overall there is continuous loss of endothel Conclusion: ial cells in all cases of Keratoplasty regardless of the indication for keratoplasty and the type of keratoplasty. Even in cases of clear grafts, there is a continuous loss of endothelial cells. It has been reported that chronic subclinical rejection, chronic low-level inammation or continued interaction between the donor endothelium and a healthy recipient endothelium could explain the greater cell loss

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Endothelial Cell Loss after Phacoemulsification according to Different Anterior Chamber Depths

Journal of Ophthalmology ◽

10.1155/2015/210716 ◽

2015 ◽

Vol 2015 ◽

pp. 1-7 ◽

Cited By ~ 10

Author(s):

Hyung Bin Hwang ◽

Byul Lyu ◽

Hye Bin Yim ◽

Na Young Lee

Keyword(s):

Endothelial Cell ◽

Anterior Chamber ◽

Corneal Thickness ◽

Cell Loss ◽

Dissipated Energy ◽

Corneal Endothelial Cells ◽

Endothelial Cell Loss ◽

Cumulative Dissipated Energy ◽

The Difference ◽

A Prospective Study

Purpose. To compare the loss of corneal endothelial cells after phacoemulsification according to different anterior chamber depths (ACDs).Methods. We conducted a prospective study on 135 eyes with senile cataracts. Eyes with nuclear density grades of 2 to 4 were divided into three groups according to ACD: ACD I, 1.5 < ACD ≤ 2.5 mm; ACD II, 2.5 < ACD ≤ 3.5 mm; or ACD III, 3.5 < ACD ≤ 4.5 mm. Intraoperative mean cumulative dissipated energy (CDE) was measured. Clinical examinations included central corneal thickness (CCT) and endothelial cell count (ECC) preoperatively and 2 months postoperatively.Results. There were no significant differences in CDE among the ACD groups (P>0.05). Endothelial cell loss was significantly higher in ACD I than in ACD III in grades 3 and 4 cataract density groups 2 months after phacoemulsification (P<0.05). There were also more changes in CCT in all of the cataract density groups in the ACD I group compared to the ACD II and III groups 2 months postoperatively, but the difference was not statistically significant.Conclusions. Eyes with shallow ACDs, especially those with relatively hard cataract densities, can be vulnerable to more corneal endothelial cell loss in phacoemulsification surgery.

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Improved Femtosecond Laser-Assisted Phacoemulsification of Hard Nucleus Cataract

Ophthalmology in Russia ◽

10.18008/1816-5095-2020-4-733-738 ◽

2020 ◽

Vol 17 (4) ◽

pp. 733-738

Author(s):

Yu. N. Yousef ◽

G. V. Voronin ◽

S. N. Yousef ◽

A. S. Vvedenskiy ◽

L. Alkharki ◽

...

Keyword(s):

Endothelial Cells ◽

Femtosecond Laser ◽

Cell Loss ◽

Corneal Endothelial Cells ◽

Effective Technique ◽

Average Loss ◽

Improved Technology ◽

Determining Factor ◽

Prostaglandin Synthesis Inhibitors ◽

Ultrasonic Load

Purpose: The development and clinical study of improved femtosecond laser-assisted phacoemulsification (PE) technology of hard nucleus cataract.Patients and methods. Improved femtosecond laser-assisted PE of hard nucleus cataract performed in 83 patients (93 eyes) (1st group), known femtosecond laser-assisted PE technique performed in 72 patients (78 eyes) (2nd group), torsional PE performed in 81 patients (89 eyes) (3rd group). The degree of intraoperative myosis, effective ultrasound time, corneal endothelial cell loss were evaluated in all groups.Results. Femtosecond laser-assisted PE is an effective technique for hard nucleus cataract removal, which can significantly reduce the energy ultrasonic load on the eye tissue. The proposed method for the instillation of prostaglandin synthesis inhibitors and the observance of the minimum possible interval between the first and second stages of the operation can prevent significant intraoperative narrowing of the pupil. A significant narrowing of the pupil by more than 2 mm after the femtolaser stage was noted in 7 (7.5 %) cases in the 1st group of patients, in 15 (16.9 %) cases in the 2nd group, in 5 (6.4 %) cases in the 3rd group. The results of the study has shown a significant decrease in the effective ultrasound time for a femtosecond laser-assisted PE compared with a torsional PE. The effective ultrasound time was in the 1st group (improved technology of femtosecond laser-assisted PE) — 3.81 ± 0.75, in the 2nd group (known technology of the femtosecond laser-assisted PE) — 5.23 ± 1.07 s (p < 0.05), in the 3rd group (OZil technology) — 8.67 ± 1.83 s (p < 0.05). The decrease in the effective ultrasound time has become a determining factor in reducing the loss of corneal endothelial cells in both femtosecond laser-assisted PE technologies compared to torsional PE. The average loss of corneal endothelial cells 3 months after surgery was 8.7 ± 1.8 % in the 1st group, 10.3 ± 2.1% in the 2nd group, 13.5 ± 2.7 % (p < 0.05) in the 3rd group of patients.Conclusion. The proposed improved technology contributes to the solution of some problems that characterize femtosecond laser-assisted PE, and also helps to reduce the effective ultrasound time and the loss of corneal endothelial cells.

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