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Angiopoietin-1 and Tie2-Based Dual Cell Therapy Enhances Antiangiogenic Barrier Function in a Retina-Mimetic Model
Cha Yeon Kim1, Cholong Jeong1, Youngjin Han2
1Department of Convergence Medicine, Asan Institute for Life Sciences, Asan Medical Center, 88 Olympic-ro 43-gil, Songpa-gu, Seoul, 05505, Republic of Korea.
Background:
Choroidal neovascularization (CNV) is a major pathological process underlying retinal degenerative diseases such as wet age-related macular degeneration. While anti-VEGF therapies are widely used, limitations in response and vascular instability necessitate new approaches that promote both antiangiogenic effects and barrier restoration.
Methods:
A dual-cell therapy strategy was developed using human umbilical vein endothelial cells (HUVECs) genetically modified to overexpress Tie2 and mesenchymal stem cells (MSCs) engineered to secrete Angiopoietin-1 (Ang1). Antiangiogenic efficacy was evaluated using scratch assays, Transwell migration, and tube formation under VEGF stimulation. A retina-mimetic 2.5D co-culture system incorporating iPSC-derived RPE cells and mCherry-labeled ECs was used to assess endothelial invasion and epithelial barrier preservation.
Results:
Tie2/Ang1-modified cells significantly suppressed angiogenic behavior. Transwell migration showed OD595 crystal violet absorbance decreased from 3.54 ± 0.27 (control HUVEC) to 1.28 ± 0.08 (Tie2 overexpressed HUVEC in MSC Ang1 conditioned medium) under VEGF stimulation (p < 0.01). Tube formation area cultured in VEGF dropped from 1.25 ± 0.05 in control group to 0.74 ± 0.07 in Tie2 overexpressed group cultured with MSC-Ang1 conditioned medium (p < 0.01). In the retina-mimetic model, EC infiltration to the RPE monolayer across Transwell membrane decreased from 52.2 ± 8.5% in control HUVEC to 5.6 ± 4.3% with HUVEC-Tie2 + Ang1 conditioned medium under VEGF (p < 0.001).
Conclusion:
This study demonstrates that co-delivery of Ang1 and Tie2 via engineered ECs and MSCs synergistically inhibits VEGF-induced angiogenesis and choroidal migration while protecting epithelial barrier function. The retina-mimetic co-culture platform further validates the translational relevance of this dual-cell approach as a regenerative and antiangiogenic strategy in retinal vascular disease.
Insights
This study developed a dual-cell therapy using engineered endothelial cells (ECs) and stem cells to inhibit choroidal neovascularization (CNV). The novel approach shows promise for treating retinal vascular diseases by restoring barrier function and reducing abnormal blood vessel growth.
Area of Science:
- Ophthalmology
- Regenerative Medicine
- Cell Therapy
Background:
- Choroidal neovascularization (CNV) is a key factor in retinal diseases like wet age-related macular degeneration.
- Current anti-VEGF therapies have limitations, necessitating new strategies for anti-angiogenesis and barrier restoration.
Purpose of the Study:
- To develop and evaluate a dual-cell therapy combining engineered human umbilical vein endothelial cells (HUVECs) overexpressing Tie2 and mesenchymal stem cells (MSCs) secreting Angiopoietin-1 (Ang1).
- To assess the antiangiogenic and barrier-protective effects of this dual-cell therapy in vitro and in a retina-mimetic model.
Main Methods:
- Genetically modified HUVECs (Tie2) and MSCs (Ang1) were used in a dual-cell therapy strategy.
- In vitro assays (scratch, Transwell migration, tube formation) evaluated antiangiogenic efficacy under VEGF stimulation.
- A 2.5D retina-mimetic co-culture system with iPSC-derived RPE cells and ECs assessed endothelial invasion and barrier preservation.
Main Results:
- The Tie2/Ang1-modified cells significantly suppressed VEGF-induced angiogenic behavior, reducing migration and tube formation.
- Endothelial cell (EC) infiltration into the retinal pigment epithelium (RPE) monolayer was drastically reduced in the retina-mimetic model.
- Quantitative data showed significant decreases in migration and tube formation in the engineered cell groups compared to controls.
Conclusions:
- Co-delivery of Ang1 and Tie2 via engineered ECs and MSCs synergistically inhibits angiogenesis and choroidal migration.
- This dual-cell therapy effectively protects epithelial barrier function, offering a regenerative and antiangiogenic strategy for retinal vascular diseases.
- The retina-mimetic co-culture model validates the translational potential of this therapeutic approach.
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