Related Experiment Video
Updated: Aug 17, 2025
07:04
Electroporation-Based Genetic Modification of Primary Human Pigment Epithelial Cells Using the Sleeping Beauty Transposon System
Published on: February 4, 2021
2.1K
Genome Editing of Pik3cd Impedes Abnormal Retinal Angiogenesis
Wenyi Wu1,2,3, Gaoen Ma4, Hui Qi5
1Department of Ophthalmology, Hunan Key Laboratory of Ophthalmology, National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, China.
Human Gene Therapy
|December 14, 2022
Summary
Gene editing of PI3Kδ (phosphoinositide 3-kinase delta) effectively reduced pathological retinal angiogenesis in a mouse model. This novel approach targets abnormal blood vessel growth implicated in diseases like PDR.
Area of Science:
- Ophthalmology
- Molecular Biology
- Genetics
Background:
- Abnormal angiogenesis, or new blood vessel formation, is a hallmark of diseases like proliferative diabetic retinopathy (PDR).
- Signaling pathways involving phosphoinositide 3-kinases (PI3Ks) are crucial regulators of angiogenesis.
- The p110δ catalytic subunit of PI3Kδ is upregulated in pathological retinal vessels.
Purpose of the Study:
- To investigate the therapeutic potential of targeting PI3Kδ for abnormal retinal angiogenesis.
- To develop and validate a CRISPR/Cas9 gene editing system to reduce PI3Kδ expression in endothelial cells.
Main Methods:
- Development of a dual recombinant adeno-associated viral (rAAV) system for delivering CRISPR/Cas9.
- Endothelial-specific expression of Streptococcus pyogenes (Sp) Cas9 using the intercellular adhesion molecule 2 promoter (pICAM2).
- Targeted genomic editing of Pik3cd, the gene encoding p110δ, in cultured endothelial cells and a mouse model of oxygen-induced retinopathy (OIR).
Main Results:
- The rAAV system achieved 80% DNA insertion/deletion in the Pik3cd locus and 70% depletion of p110δ expression in vitro.
- In the OIR mouse model, Pik3cd editing significantly decreased p110δ expression and Akt activation.
- CRISPR/Cas9-mediated Pik3cd editing markedly reduced pathological retinal angiogenesis in OIR mice.
Conclusions:
- Genomic editing of Pik3cd represents a novel strategy for treating abnormal retinal angiogenesis.
- Targeting PI3Kδ pathway via gene editing offers a potential therapeutic avenue for PDR and related vascular diseases.
