Rapid generation of functional vascular organoids via simultaneous transcription factor activation of endothelial and mural lineages

Area of Science:

• Stem cell biology
• Vascular biology
• Regenerative medicine

Background:

• Vascular organoids (VOs) are crucial for studying vascular development and disease.
• Independent control of endothelial and mural cells in VOs is a significant challenge.

Purpose of the Study:

• To develop a streamlined method for generating functional vascular organoids (VOs) from induced pluripotent stem cells (iPSCs).
• To enable independent control over endothelial and mural cell differentiation within VOs.
• To establish a versatile platform for vascular modeling and regenerative therapy.

Main Methods:

• Utilized orthogonal activation of transcription factors ETV2 and NKX3.1 via Dox-inducible or modRNA systems.
• Generated 3D VOs from iPSCs within 5 days without extracellular matrix (ECM) embedding.
• Analyzed VO development using single-cell RNA sequencing and in vivo engraftment studies.

Main Results:

• Successfully generated functional 3D VOs with co-differentiated endothelial cells (iECs) and mural cells (iMCs).
• Demonstrated temporal regulation of TF expression modulated iEC phenotypes (arterial, angiogenic).
• Showcased in vivo functionality: VOs engrafted, formed perfused vasculature, and promoted revascularization in disease models.

Conclusions:

• Established a rapid and versatile vascular organoid platform.
• The method allows for precise control over vascular cell types and phenotypes.
• VOs show significant potential for vascular disease modeling and cell-based regenerative therapies.