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The Transcription Factor RUNX1 Aggravates Hypoxia-Induced Human Retinal Microvascular Endothelial Cell Dysfunction by
Yanli Zhang1,2, Shuang Qiu3, Lingyan Liu3
1Department of Ophthalmology, Zhongshan Aier Eye Hospital, Zhongshan, Guangdong, China yanliZhangdoctor@hotmail.com.
Insights
Runt-related transcription factor 1 (RUNX1) suppresses Trefoil factor 1 (TFF1) expression, worsening hypoxia-induced dysfunction in human retinal vascular endothelial cells (HRVECs). This finding is crucial for understanding retinopathy of prematurity (ROP).
Area of Science:
- Ophthalmology
- Vascular Biology
- Molecular Biology
Background:
- Retinopathy of prematurity (ROP) is a retinal vaso-proliferative disorder causing vision loss in premature infants.
- Neovascularization and angiogenesis are key pathological features of ROP.
- Trefoil factor 1 (TFF1) is implicated in retinal angiogenesis in diabetic retinopathy, but its role in ROP is unclear.
Purpose of the Study:
- To investigate the regulatory role of TFF1 in hypoxia-induced human retinal vascular endothelial cells (HRVECs).
- To elucidate the underlying molecular mechanisms of TFF1's function in an in vitro ROP model.
Main Methods:
- Established in vitro ROP models using hypoxia-exposed HRVECs.
- Assessed cell viability, migration, and angiogenesis using CCK-8, wound healing, and tube formation assays.
- Quantified TFF1 and RUNX1 expression via RT-qPCR and Western blotting; confirmed RUNX1 binding to the TFF1 promoter using ChIP and luciferase assays.
Main Results:
- Hypoxia decreased TFF1 and increased RUNX1 expression in HRVECs.
- Hypoxia enhanced HRVEC viability, migration, and angiogenesis, effects counteracted by TFF1 upregulation or RUNX1 knockdown.
- RUNX1 directly bound to the TFF1 promoter, transcriptionally repressing TFF1 expression.
Conclusions:
- RUNX1 transcriptionally suppresses TFF1, exacerbating hypoxia-induced HRVEC dysfunction.
- TFF1 acts as a protective factor against ROP-related vascular changes.
- Targeting the RUNX1-TFF1 pathway may offer therapeutic strategies for ROP.
Objective:
As a retinal vaso-proliferative disorder, retinopathy of prematurity (ROP) is characterized by neovascularization and angiogenesis, causing irreversible retinal damage and even visual loss among premature infants. Trefoil factor 1 (TFF1) has been identified as a key regulator in mediating retinal angiogenesis in diabetic retinopathy. However, whether TFF1 can mediate the angiogenic process in ROP remains unknown. Here, we aimed to investigate the regulatory function of TFF1 and its underlying mechanisms in hypoxia-exposed human retinal vascular endothelial cells (HRVECs) in vitro.
Methods:
HRVECs were exposed to hypoxia condition to establish the in vitro ROP models. HRVEC viability was validated using CCK-8 assay. The migratory and angiogenic capacities of HRVECs were assessed by wound healing and tube formation assays, respectively. RT-qPCR was performed to detect gene levels. Western blotting was used to measure the protein levels of TFF1 and Runt-related transcription factor 1 (RUNX1). The binding relationship between RUNX1 to TFF1 promoter was confirmed by chromatin immunoprecipitation and luciferase reporter assays.
Results:
Hypoxia downregulated TFF1 expression and elevated RUNX1 expression in HRVECs. Moreover, hypoxic condition increased HRVEC viability and accelerated HRVEC migration and angiogenesis, which were antagonized by TFF1 elevation or RUNX1 knockdown. RUNX1 as a transcription factor bound to TFF1 promoter and transcriptionally repressed TFF1 expression in HRVECs. In rescue assays, overexpression of TFF1 counteracted the promotive effect of RUNX1 overexpression on the viability, migratory and angiogenic abilities of HRVECs under hypoxia.
Conclusions:
RUNX1 transcriptionally suppresses TFF1 expression to aggravate hypoxia-induced HRVEC dysfunction.
