Perturbations of Glutathione and Sphingosine Metabolites in Port Wine Birthmark Patient-Derived Induced Pluripotent Stem Cells

Vi Nguyen; Jacob Kravitz; Chao Gao; Marcelo L Hochman; Dehao Meng; Dongbao Chen; Yunguan Wang; Anil G Jegga; J Stuart Nelson; Wenbin Tan

doi:10.3390/metabo13090983

Perturbations of Glutathione and Sphingosine Metabolites in Port Wine Birthmark Patient-Derived Induced Pluripotent Stem Cells

Metabolites. 2023 Aug 31;13(9):983. doi: 10.3390/metabo13090983.

Authors

Vi Nguyen¹, Jacob Kravitz¹, Chao Gao¹, Marcelo L Hochman^{2

3}, Dehao Meng⁴, Dongbao Chen⁵, Yunguan Wang^{6

7

8}, Anil G Jegga^{6

9}, J Stuart Nelson¹⁰, Wenbin Tan^{1

11}

Affiliations

¹ Department of Cell Biology and Anatomy, School of Medicine, University of South Carolina, Columbia, SC 29209, USA.
² The Facial Surgery Center and the Hemangioma & Malformation Treatment Center, Charleston, SC 29425, USA.
³ Department of Otolaryngology-Head and Neck Surgery, Medical University of South Carolina, Charleston, SC 29425, USA.
⁴ Applied Physics Program, California State University San Marcos, San Marcos, CA 92096, USA.
⁵ Department of Obstetrics and Gynecology, University of California, Irvine, CA 92617, USA.
⁶ Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH 45229, USA.
⁷ Division of Gastroenterology, Cincinnati Children Hospital Medical Center, Cincinnati, OH 45229, USA.
⁸ Division of Human Genetics, Cincinnati Children Hospital Medical Center, Cincinnati, OH 45229, USA.
⁹ Division of Biomedical Informatics, Cincinnati Children Hospital Medical Center, Cincinnati, OH 45229, USA.
¹⁰ Departments of Surgery and Biomedical Engineering, Beckman Laser Institute and Medical Clinic, University of California, Irvine, CA 92617, USA.
¹¹ Department of Biomedical Engineering, College of Engineering and Computing, University of South Carolina, Columbia, SC 29208, USA.

Abstract

Port Wine Birthmarks (PWBs) are a congenital vascular malformation on the skin, occurring in 1-3 per 1000 live births. We have recently generated PWB-derived induced pluripotent stem cells (iPSCs) as clinically relevant disease models. The metabolites associated with the pathological phenotypes of PWB-derived iPSCs are unknown, and so we aim to explore them in this study. Metabolites were separated by ultra-performance liquid chromatography and screened with electrospray ionization mass spectrometry. Orthogonal partial least-squares discriminant, multivariate, and univariate analyses were used to identify differential metabolites (DMs). KEGG analysis was used to determine the enrichment of metabolic pathways. A total of 339 metabolites was identified. There were 22 DMs, among which nine were downregulated-including sphingosine-and 13 were upregulated, including glutathione in PWB iPSCs, as compared to controls. Pathway enrichment analysis confirmed the upregulation of glutathione and the downregulation of sphingolipid metabolism in PWB-derived iPSCs as compared to normal ones. We next examined the expression patterns of the key molecules associated with glutathione metabolism in PWB lesions. We found that hypoxia-inducible factor 1α (HIF1α), glutathione S-transferase Pi 1 (GSTP1), γ-glutamyl transferase 7 (GGT7), and glutamate cysteine ligase modulatory subunit (GCLM) were upregulated in PWB vasculatures as compared to blood vessels in normal skin. Other significantly affected metabolic pathways in PWB iPSCs included pentose and glucuronate interconversions; amino sugar and nucleotide sugars; alanine, aspartate, and glutamate; arginine, purine, D-glutamine, and D-glutamate; arachidonic acid, glyoxylate, and dicarboxylate; nitrogen, aminoacyl-tRNA biosynthesis, pyrimidine, galactose, ascorbate, and aldarate; and starch and sucrose. Our data demonstrated that there were perturbations in sphingolipid and cellular redox homeostasis in PWB vasculatures, which could facilitate cell survival and pathological progression. Our data implied that the upregulation of glutathione could contribute to laser-resistant phenotypes in some PWB vasculatures.

Keywords: Port Wine Birthmark; glutathione; hypoxia; induced pluripotent stem cells; metabolome; sphingolipid; sphingosine.

Abstract

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