Transient Vascular Cell Integration and Intrinsic Media-Stimulated Endothelial Activation Reveal Diabetic Retinopathy Signatures in Human Retinal Organoids
- 1newcastle university
Abstract
Retinal organoids (ROs) model human retinogenesis but remain avascular and lack immune components, limiting their utility for studying vascular retinopathies. We generated vascular organoids (VOrs) from human pluripotent stem cells and evaluated their integration into developing ROs. Dissociated VOr cells (day 12 or day 30) were seeded onto 2D retinal neuroepithelium before 3D transition, followed by long-term culture to day 210. VOr derived cells migrated into ROs and transiently persisted. V30 ROs preserved retinal lamination, major neuronal subtypes and light evoked retinal ganglion cell responses comparable to non vascularised controls, whereas media-only ROs cultured in proangiogenic conditions showed structural disorganisation and reduced neuronal survival. Single-cell RNA sequencing revealed an endogenous, inflammatory endothelial-like population with partial endothelial-to-mesenchymal features and a perivascular fibroblast-like cluster in pro-angiogenic media only RO controls. The endothelial-like cell cluster was significantly enriched for genes upregulated in human retinal endothelial cells exposed to high glucose, indicating a diabetic retinopathy like transcriptional state. Microglia like and border associated macrophage populations were present across all conditions, with developmental shifts in morphology and marker expression. Our findings define conditions under which vascular cells can be incorporated into human ROs without disrupting retinal architecture, while uncovering an intrinsic capacity for inflammatory endothelial activation that partially recapitulates diabetic retinopathy. This work advances the engineering of vascularised, immunocompetent retinal models and highlights endogenous vascular immune programmes that must be considered when modelling retinal disease in vitro.
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BibTeX
@misc{tsikandelova2026transient,
title = {{Transient Vascular Cell Integration and Intrinsic Media-Stimulated Endothelial Activation Reveal Diabetic Retinopathy Signatures in Human Retinal Organoids}},
author = {R. Tsikandelova and R. Schettler and M. D. Teasdale and J. Kelk and V. Boczonadi and B. Dorgau and R. Hussain and J. M. Coxhead and R. Medina and S. White and R. Zweigerdt and L. Armstrong and G. Hilgen and M. Lako},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.01.755879},
url = {https://www.biorxiv.org/content/10.64898/2026.10.01.755879v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - Transient Vascular Cell Integration and Intrinsic Media-Stimulated Endothelial Activation Reveal Diabetic Retinopathy Signatures in Human Retinal Organoids
AU - R. Tsikandelova
AU - R. Schettler
AU - M. D. Teasdale
AU - J. Kelk
AU - V. Boczonadi
AU - B. Dorgau
AU - R. Hussain
AU - J. M. Coxhead
AU - R. Medina
AU - S. White
AU - R. Zweigerdt
AU - L. Armstrong
AU - G. Hilgen
AU - M. Lako
PY - 2026
DA - 2026/10/01
PB - bioRxiv
DO - 10.64898/2026.10.01.755879
UR - https://www.biorxiv.org/content/10.64898/2026.10.01.755879v1
AB - Retinal organoids (ROs) model human retinogenesis but remain avascular and lack immune components, limiting their utility for studying vascular retinopathies. We generated vascular organoids (VOrs) from human pluripotent stem cells and evaluated their integration into developing ROs. Dissociated VOr cells (day 12 or day 30) were seeded onto 2D retinal neuroepithelium before 3D transition, followed by long-term culture to day 210. VOr derived cells migrated into ROs and transiently persisted. V30 ROs preserved retinal lamination, major neuronal subtypes and light evoked retinal ganglion cell responses comparable to non vascularised controls, whereas media-only ROs cultured in proangiogenic conditions showed structural disorganisation and reduced neuronal survival. Single-cell RNA sequencing revealed an endogenous, inflammatory endothelial-like population with partial endothelial-to-mesenchymal features and a perivascular fibroblast-like cluster in pro-angiogenic media only RO controls. The endothelial-like cell cluster was significantly enriched for genes upregulated in human retinal endothelial cells exposed to high glucose, indicating a diabetic retinopathy like transcriptional state. Microglia like and border associated macrophage populations were present across all conditions, with developmental shifts in morphology and marker expression. Our findings define conditions under which vascular cells can be incorporated into human ROs without disrupting retinal architecture, while uncovering an intrinsic capacity for inflammatory endothelial activation that partially recapitulates diabetic retinopathy. This work advances the engineering of vascularised, immunocompetent retinal models and highlights endogenous vascular immune programmes that must be considered when modelling retinal disease in vitro.
ER -