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Series GSE139221 Query DataSets for GSE139221
Status Public on Oct 22, 2019
Title Modeling of aniridia-related keratopathy by CRISPR/Cas9 genome editing of human limbal epithelial cells and rescue by recombinant PAX6 protein
Organism Homo sapiens
Experiment type Expression profiling by high throughput sequencing
Summary Heterozygous PAX6 gene mutations leading to haploinsufficiency are the main cause of congenital aniridia, a rare and progressive panocular disease characterized by reduced visual acuity. Up to 90% of patients suffer from aniridia-related keratopathy (ARK), caused by a combination of factors including limbal epithelial stem-cell (LSC) deficiency, impaired healing response and abnormal differentiation. It usually begins in the first decade of life, resulting in recurrent corneal erosions, sub-epithelial fibrosis and corneal opacification. Unfortunately, there are currently no efficient treatments available for these patients and no in vitro model for this pathology. We used CRISPR/Cas9 technology to introduce into the PAX6 gene of LSCs a heterozygous nonsense mutation found in ARK patients. Nine clones carrying a p.E109X mutation on one allele were obtained with no off-target mutations. Compared to the parental WT-LSCs, heterozygous mutant LSCs displayed reduced expression of PAX6 and marked slow-down of cell proliferation, migration and detachment. Remarkably, addition to the culture medium of recombinant PAX6 protein fused to a cell penetrating peptide was able to activate the endogenous PAX6 gene and to rescue phenotypic defects of mutant LSCs, suggesting that administration of such recombinant PAX6 protein could be a promising therapeutic approach of congenital aniridia. More generally, our results demonstrate that introduction of disease mutations into LSCs by CRISPR/Cas9 genome editing allows creating relevant cellular models of ocular disease that should greatly facilitate screening of novel therapeutic approaches.
Overall design Genome editing of aniridia-related keratopathy and rescue. Gene expression comparison between normal and genome-edited PAX6 mutation containing T-limba cells
Contributor(s) Roux LN, Petit I, Domart R, Concordet J, Qu J, Zhou H, Joliot A, Ferrigno O, Aberdam D
Citation(s) 29808941
Submission date Oct 21, 2019
Last update date Jun 14, 2022
Contact name Jo Huiqing Zhou
Organization name Radboud University
Street address Geert Grooteplein 26/18
City Nijmegen
ZIP/Postal code 6525GA
Country Netherlands
Platforms (1)
GPL16791 Illumina HiSeq 2500 (Homo sapiens)
Samples (4)
GSM4134291 Limbal-TERT-wt-34-12594
GSM4134292 Limbal-TERT-wt-35-12595
GSM4134293 Limbal-TERT-cloneW-49-12596
BioProject PRJNA578775
SRA SRP226540

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Supplementary file Size Download File type/resource
GSE139221_CountData.txt.gz 366.3 Kb (ftp)(http) TXT
GSE139221_DEG_TableX.xlsx 45.2 Kb (ftp)(http) XLSX
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Raw data are available in SRA
Processed data are available on Series record

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