Date of Award
Summer 2025
Document Type
Open Access Dissertation
Department
Biomedical Science
First Advisor
Wenbin Tan
Abstract
Congenital capillary malformation (CM) is a cutaneous vascular defect that occurs in around 0.3-0.5% of births. The lesion appears pink or red macule at birth, and it progressively darkens to purple in adulthood with soft tissue hypertrophy and a development of vascular nodules. The current treatment for CM is Pulse Dye Laser and photodynamic therapy (PDT) with marginal outcomes due to reoccurrence and laser treatment resistance. Lack of clinically relevant models has substantially hampered CM mechanistic and drug screening studies. Utilizing induced pluripotent stem cells (iPSCs), we generated patient-derived iPSCs and differentiated them into induced endothelial cells (iECs) and vascular organoids (VO). CM iECs recapitulate CM-like phenotypes, such as the formation of large-sized vasculature, both in vitro and in vivo. Single-cell RNA sequencing revealed that CM iEC had an attenuated iEC lineage differentiation but enhancement in induced smooth muscle cell (iSMC) lineage. Using single vasculature spatial whole transcriptome atlas, we demonstrated prominent phenotypes of CM vasculature including impaired adherens junctions (AJs), discorded expressions of vascular lineage biomarkers such as upregulation of nuclear NR2F2, and enhanced endothelial-to-mesenchymal transition (EndMT). CM phosphoproteomic profiles revealed that differentially phosphorylated proteins converged functional enrichments of AJs and EndMT. When NR2F2 over-expression was induced at the progenitor cell stage during the differentiation course of iPSCs, CM iECs showed downregulation of endothelial biomarkers but upregulation of SMC biomarkers. Our data suggests NR2F2 promotes progenitor cell-derived EndMT in CM. Additionally, VOs demonstrated the self-assembly of differentiated vascular lineages from iECs and iSMCs. The iECs and iSMCs juxtaposed to form vascular branches in VOs. CM patient iPSC-derived VOs showed a higher density of endothelial and SMC populations and greater vascular branch lengths as compared with VOs derived from iPSCs generated from healthy skin biopsies. Moreover, CM iECs exhibited laser resistance in response to PDT treatment compared with normal iECs. CM iECs with laser resistance were enriched dysregulated pathways of hypoxia, ROS, oxidative phosphorylation, and negative regulation of apoptosis. They had increased glutathione production, thus can survive elevated ROS. Through data-driven drug repurposing, we identified several compounds, including sitagliptin, liarozole, and xanthohumol, that could sensitize CM iECs with laser resistance.
Rights
© 2025, Vi Bich Nguyen
Recommended Citation
Nguyen, V. B.(2025). Harnessing Induced Pluripotent Stem Cell as a Pivotal Disease Model for Capillary Malformation. (Doctoral dissertation). Retrieved from https://scholarcommons.sc.edu/etd/8494