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Updated: Apr 18, 2026
An Ex Vivo Tissue Culture Model for Fibrovascular Complications in Proliferative Diabetic Retinopathy
Published on: January 25, 2019
Microarray analysis of gene expression in fibrovascular membranes excised from patients with proliferative diabetic
Keijiro Ishikawa1, Shigeo Yoshida1, Yoshiyuki Kobayashi1
1Department of Ophthalmology, Kyushu University Graduate School of Medical Sciences, Fukuoka, Japan.
Purpose:
We determined the profile of genes expressed in fibrovascular membranes (FVMs).
Methods:
Six FVMs were surgically removed from patients with proliferative diabetic retinopathy (PDR) during pars plana vitrectomy with membrane peeling. The FVMs were classified into three active FVMs or three inactive FVMs according to the presence or absence of neovascularization (NV) in the membranes. Total RNA was isolated from the six FVMs and also from three normal human retinas. The DNA microarray analysis was performed to compare the genes expressed in the FVMs to those in normal human retinas, and also between active and inactive FVMs. Ingenuity pathway analysis (IPA) was used to determine the key biological networks related to the genes that were significantly altered. Quantitative RT-PCR and immunohistochemistry were performed to validate the microarray analyses.
Results:
There were 87 genes expressed at significantly higher levels in FVMs than in normal human retinas. Functional classification of these genes showed that the most clustered genes were those related to extracellular matrix formation. The top biological network generated by the IPA was cellular assembly and organization involving nodes of genes related to extracellular matrix formation. These networks included the collagen family and matricellular proteins, THBS2, POSTN, and TNC. There were 91 genes significantly upregulated in active FVMs, and the most clustered functional category was angiogenesis. In contrast, 89 genes were significantly upregulated in inactive FVMs, and the most clustered functional category was metabolism. The IPA revealed that the top biological network related to the genes that were significantly altered in this comparison was cell-to-cell signaling, and interactions involving the PDGF and TGFβ families. The results of quantitative RT-PCR analyses and immunohistochemistry for several selected molecules were in good agreement with the microarray data.
Conclusions:
Our data indicate that extracellular matrix-related molecules such as POSTN, TNC, TGFβ, and angiogenic factors have important roles in promoting the development of FVMs associated with PDR.
Insights
Gene expression in fibrovascular membranes (FVMs) from proliferative diabetic retinopathy (PDR) patients reveals key roles for extracellular matrix molecules and angiogenic factors in FVM development.
Area of Science:
- Ophthalmology
- Molecular Biology
- Genomics
Background:
- Proliferative diabetic retinopathy (PDR) is a severe complication of diabetes.
- Fibrovascular membranes (FVMs) are a hallmark of PDR, leading to vision loss.
- Understanding the molecular mechanisms underlying FVM formation is crucial for developing targeted therapies.
Purpose of the Study:
- To profile gene expression in fibrovascular membranes (FVMs) associated with PDR.
- To compare gene expression between active and inactive FVMs.
- To identify key biological networks and molecular players involved in FVM development.
Main Methods:
- Surgical removal of FVMs from PDR patients.
- Classification of FVMs into active and inactive groups based on neovascularization.
- DNA microarray analysis to compare gene expression profiles.
- Ingenuity pathway analysis (IPA) for network identification.
- Validation using quantitative RT-PCR and immunohistochemistry.
Main Results:
- 87 genes were significantly upregulated in FVMs compared to normal retinas, primarily related to extracellular matrix formation.
- Active FVMs showed upregulation of 91 genes involved in angiogenesis.
- Inactive FVMs exhibited upregulation of 89 genes related to metabolism.
- Key networks involved extracellular matrix formation, cellular assembly, and cell-to-cell signaling.
Conclusions:
- Extracellular matrix molecules (e.g., POSTN, TNC) are significantly involved in FVM development.
- Angiogenic factors play a critical role in the progression of FVMs in PDR.
- TGFβ signaling pathways are implicated in the pathogenesis of FVMs.
