Vi Nguyen

All Publications

• Vascular Organoids Derived from Capillary malformation-induced Pluripotent Stem Cells Exhibit Disease-Relevant Phenotypes STEM CELL REVIEWS AND REPORTS Nguyen, V., Harper, A., Azuero, M., Castellanos, I., He, S., Hochman, M. L., Wenceslau, C. F., Chen, D., Jegga, A. G., Wang, Y., Fan, D., Nelson, J., Tan, W. 2025

  Abstract

  Capillary malformation (CM) is a congenital vascular anomaly that affects the skin, mucosa, eye, and brain. A major obstacle to mechanistic and drug screening studies for CM has been the lack of preclinical models. In this study, we established vascular organoids (VOs) generated through the self-assembly of vascular lineages of endothelial cells and smooth muscle cells differentiated from CM-induced pluripotent stem cells (iPSC). Within these VOs induced endothelial cells and smooth muscle cells organized into juxtapositions to form vascular branches. CM patient iPSC-derived VOs showed a higher density of endothelial and smooth muscle cell populations and greater vascular branch lengths as compared with VOs derived from iPSCs generated from healthy skin biopsies. Overall, this study represents the first disease-relevant VO model of CM, providing a valuable platform for future mechanistic studies and drug screening.

  View details for DOI 10.1007/s12015-025-10984-8

  View details for Web of Science ID 001578683800001

  View details for PubMedID 40996655

  View details for PubMedCentralID 5110233

• Single-vessel transcriptome map pathological landscapes and reveal NR2F2-mediated smooth muscle cell phenotype acquisition in capillary malformations. bioRxiv : the preprint server for biology Nguyen, V., Mao, I., He, S., Castellanos, I., Azuero, M., Hochman, M. L., Rong, Y., Pernomian, L., Chen, E. H., Friedman, H. I., Chen, Y., Lu, Q., Fan, D., Wenceslau, C. F., Chen, D., Nelson, J. S., Jegga, A. G., Wang, Y., Tan, W. 2025

  Abstract

  Background: Capillary malformation (CM) is a congenital vascular anomaly affecting the skin, mucosa, and brain, yet the understanding of its vascular pathogenesis remains limited.Methods: We applied spatial whole-transcriptome profiling (GeoMx) and gene set enrichment analysis within CM lesions at single vasculature level. Differentially expressed genes were validated by immunofluorescence staining. Phosphoproteomics was profiled to uncover lesion-wide phosphorylation sites on proteins. Single-cell RNA sequencing was performed on CM-derived induced pluripotent stem cells (iPSCs) to determine differentiation trajectories of lesional vascular lineages. In silico gene perturbation was used to predict candidate genes for modulating vascular pathological progression, followed by functional validation in CM iPSC-derived endothelial cells (ECs) using a Tet-on system.Results: A spatial transcriptomic atlas was constructed, and pathological landscape of individual CM vasculature was delineated. CM vessels exhibited hallmarks of endothelial-to-mesenchymal transition (EndMT), including disruption of adherens junctions (AJs), vascular identity transitions, and metabolic remodeling. Phosphoproteomics confirmed that differentially phosphorylated proteins were enriched in EndMT- and AJ-related pathways. Aberrant expression of venous transcriptional factor NR2F2 was observed in lesional ECs and correlated with progressive enlargement from capillaries to larger-caliber vessels containing multiple layers of smooth muscle cells (SMCs). In CM iPSCs, differentiation course yielded reduced ECs but increased SMCs. In silico knockout simulation predicted NR2F2 as a crucial regulator of facilitating SMC phenotype in CM. Consistently, enforced NR2F2 expression during iPSC differentiation suppressed endothelial markers while inducing SMC-associated genes.Conclusions: Single CM vasculature displays pathological hallmarks characterized by EndMT and AJ disruption, leading to progressive vascular remodeling. NR2F2 functions as a central regulatory factor orchestrating the acquisition of the SMC phenotype, thereby representing a potential therapeutic target in CM.

  View details for DOI 10.1101/2025.09.02.673874

  View details for PubMedID 40950147

• Single-Short Partial Reprogramming of the Endothelial Cells Decreases Blood Pressure via Attenuation of EndMT in Hypertensive Mice. Circulation research Pernomian, L., Waigi, E. W., Nguyen, V., Mohammed, A. D., Costa, T. J., Fontes, M. T., Kubinak, J. L., Aitken, A. V., Biancardi, V. C., Gleason, K., Shazly, T., Sinclair, D. A., McCarthy, C. G., Wang, Y., Tan, W., Wenceslau, C. F. 2025

  Abstract

  BACKGROUND: Small artery remodeling and endothelial dysfunction are hallmarks of hypertension. Evidence supports a likely causal association between cardiovascular diseases and endothelial-to-mesenchymal transition, a cellular transdifferentiation process in which endothelial cells (ECs) partially lose their identity and acquire mesenchymal phenotypes. EC reprogramming represents an innovative strategy in regenerative medicine to prevent deleterious effects induced by cardiovascular diseases.METHODS: Using partial reprogramming of ECs, via overexpression of Oct-3/4-Sox-2-Klf-4 (OSK) transcription factors, we aimed to bring ECs back to a youthful phenotype in hypertension. Primary ECs were infected with lentiviral vectors (LVs) containing the specific EC promoter Cdh5 (cadherin-5) and the reporter EGFP (enhanced green fluorescent protein) with empty vector (LV control) or LV with Oct-3/4-Sox-2-Klf-4. Confocal microscopy and Western blotting analysis were used to confirm OSK overexpression. Cellular migration, senescence, and apoptosis were evaluated. Human aortic ECs from normotensive patients and patients with hypertension were analyzed after OSK treatments for eNOS (endothelial nitric oxide synthase), NO, and genetic profile. Male and female normotensive (blood pressure normal mouse strain) and hypertensive (blood pressure high mouse strain) mice were treated with LV control or LV with Oct-3/4-Sox-2-Klf-4 and evaluated 10 days post-infection. The blood pressure, cardiac function, vascular reactivity of small arteries, and endothelial-to-mesenchymal transition inhibition were analyzed.RESULTS: OSK overexpression induced partial EC reprogramming in vitro, and these cells had lower migratory capability. OSK treatment of blood pressure high mouse strain mice reduced blood pressure and resistance arteries hypercontractility, via the attenuation of endothelial-to-mesenchymal transition and elastin breaks. EGFP was detected in vivo in the prefrontal cortex. OSK-treated hypertensive human aortic ECs showed high eNOS activation and NO production, with low reactive oxygen species formation. Single-cell RNA analysis showed that OSK alleviated EC senescence and endothelial-to-mesenchymal transition, restoring their phenotypes in human aortic ECs from patients with hypertension.CONCLUSIONS: Overall, these data indicate that OSK treatment and EC reprogramming can decrease blood pressure and reverse hypertension-induced vascular damage.

  View details for DOI 10.1161/CIRCRESAHA.124.324909

  View details for PubMedID 40899274