CircTHADA regulates endothelial cell pyroptosis in diabetic retinopathy through miR-494-3p/CASP1/GSDMD-N/IL-1β
Shuai He1, Chufeng Gu2, Chunren Meng3
1School of Life Sciences, Westlake University, Westlake Laboratory of Life Sciences and Biomedicine, Institute of Basic Medical Science, Westlake Institute for Advanced Study, Hangzhou, Zhejiang, PR China.
Abstract:
Our study aimed to elucidate the mechanism by which circTHADA competitively adsorbs miR-494-3p to regulate CASP1-mediated endothelial cell (EC) pyroptosis in diabetic retinopathy (DR). To be specific, we used high glucose (HG)-induced human retinal microvascular endothelial cells (HRMECs) as DR cell models and streptozotocin (STZ)-treated mice as DR mouse models. The expression levels of circTHADA, miR-494-3p, CASP1, NLRP3, GSDMD-N and IL-1β were detected and flow cytrometry was applied to measure cell pyroptosis rate and dual luciferase reporter assays were utilized to determine the direct binding sites. As a result, exacerbated EC pyroptosis in DR was detected in DR cell and mouse models. Based on differentially expressed circRNA profiles by microarray and experimental verification, circTHADA was filtered and identified to regulate CASP1-mediated EC pyroptosis. miR-494-3p was then proven to be involved in circTHADA-mediated ceRNA network by bioinformatics analysis and experimental verification. Further gain- and loss-of-function experiments and rescue experiments revealed the function of the circTHADA/miR-494-3p/CASP1 axis in pyroptosis.
Insights
Circular RNA THADA (circTHADA) exacerbates diabetic retinopathy by promoting endothelial cell pyroptosis through the circTHADA/miR-494-3p/CASP1 pathway. This finding reveals a novel mechanism in diabetic retinopathy progression.
Area of Science:
- Ophthalmology
- Molecular Biology
- Cell Biology
Background:
- Diabetic retinopathy (DR) is a microvascular complication of diabetes.
- Endothelial cell (EC) pyroptosis contributes to DR pathogenesis.
- The molecular mechanisms underlying EC pyroptosis in DR require further elucidation.
Purpose of the Study:
- To investigate the role of circular RNA THADA (circTHADA) in regulating EC pyroptosis in diabetic retinopathy (DR).
- To elucidate the mechanism involving circTHADA, miR-494-3p, and CASP1 in DR.
- To identify potential therapeutic targets for DR.
Main Methods:
- Established high glucose (HG)-induced human retinal microvascular endothelial cells (HRMECs) and streptozotocin (STZ)-treated mice as DR models.
- Quantified expression levels of circTHADA, miR-494-3p, CASP1, NLRP3, GSDMD-N, and IL-1β.
- Utilized flow cytometry for pyroptosis assessment and dual luciferase reporter assays for binding site verification.
- Performed microarray analysis for circRNA profiling and bioinformatics analysis for ceRNA network identification.
Main Results:
- Confirmed exacerbated EC pyroptosis in DR cell and mouse models.
- Identified circTHADA as a regulator of CASP1-mediated EC pyroptosis.
- Demonstrated that miR-494-3p is involved in the circTHADA-mediated competing endogenous RNA (ceRNA) network.
- Established the functional axis of circTHADA/miR-494-3p/CASP1 in regulating pyroptosis through gain/loss-of-function and rescue experiments.
Conclusions:
- circTHADA promotes EC pyroptosis in DR by sponging miR-494-3p, leading to increased CASP1 expression.
- The circTHADA/miR-494-3p/CASP1 axis plays a critical role in DR pathogenesis.
- Targeting this axis may offer a novel therapeutic strategy for diabetic retinopathy.
