Single-cell profiling reveals transcriptomic signatures of vascular endothelial cells in non-healing diabetic foot ulcers

Yangzhou Lu; Xiaogang Liu; Jingling Zhao; Fan Bie; Yiling Liu; Julin Xie; Peng Wang; Junyou Zhu; Yahui Xiong; Shitian Qin; Fan Yang; Lei Chen; Yingbin Xu

doi:10.3389/fendo.2023.1275612

Single-cell profiling reveals transcriptomic signatures of vascular endothelial cells in non-healing diabetic foot ulcers

Front Endocrinol (Lausanne). 2023 Dec 1:14:1275612. doi: 10.3389/fendo.2023.1275612. eCollection 2023.

Authors

Yangzhou Lu^#^{1

2

3}, Xiaogang Liu^#^{1

2

3}, Jingling Zhao^{1

2

3}, Fan Bie^{1

2

3}, Yiling Liu^{1

2

3}, Julin Xie^{1

2

3}, Peng Wang^{1

2

3}, Junyou Zhu^{1

2

3}, Yahui Xiong^{1

2

3}, Shitian Qin^{1

2

3}, Fan Yang^{1

2

3}, Lei Chen^{1

2

3}, Yingbin Xu^{1

2

3}

Affiliations

¹ Department of Burn, Wound Repair & Reconstruction, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, Guangdong, China.
² Guangdong Provincial Engineering Technology Research Center of Burn and Wound Accurate Diagnosis and Treatment Key Technology and Series of Products, Sun Yat-Sen University, Guangzhou, Guangdong, China.
³ Institute of Precision Medicine, The First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, Guangdong, China.

^# Contributed equally.

Abstract

Background: The treatment of diabetic foot ulcers (DFUs) poses a challenging medical problem that has long plagued individuals with diabetes. Clinically, wounds that fail to heal for more than 12 weeks after the formation of DFUs are referred to as non-healing/chronic wounds. Among various factors contributing to the non-healing of DFUs, the impairment of skin microvascular endothelial cell function caused by high glucose plays a crucial role. Our study aimed to reveal the transcriptomic signatures of non-healing DFUs endothelial cells, providing novel intervention targets for treatment strategies.

Methods: Based on the GEO dataset (GSE165816), we selected DFU-Healer, DFU-Non-healer, and healthy non-diabetic controls as research subjects. Single-cell RNA transcriptomic sequencing technology was employed to analyze the heterogeneity of endothelial cells in different skin tissue samples and identify healing-related endothelial cell subpopulations. Immunofluorescence was applied to validate the sequencing results on clinical specimens.

Results: The number of endothelial cells and vascular density showed no significant differences among the three groups of skin specimens. However, endothelial cells from non-healing DFUs exhibited apparent inhibition of angiogenesis, inflammation, and immune-related signaling pathways. The expression of CCND1, ENO1, HIF1α, and SERPINE1 was significantly downregulated at the transcriptomic and histological levels. Further analysis demonstrated that healing-related endothelial cell subpopulations in non-healing DFUs has limited connection with other cell types and weaker differentiation ability.

Conclusion: At the single-cell level, we uncovered the molecular and functional specificity of endothelial cells in non-healing DFUs and highlighted the importance of endothelial cell immune-mediated capability in angiogenesis and wound healing. This provides new insights for the treatment of DFUs.

Keywords: angiogenesis; diabetic foot ulcers; immune; inflammation; non-healing wounds; single-cell RNA sequencing; vascular endothelial cell.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Diabetes Mellitus*
Diabetic Foot*
Endothelial Cells / metabolism
Gene Expression Profiling
Humans
Skin / metabolism
Wound Healing / genetics

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. The study was supported by National Natural Science Foundation of China (81772089 to YBX, 82272297 to LC, 82072179 to JLZ, and 82102332 to JYZ) and “XinRui” Talent program of First Affiliated Hospital of Sun Yat-Sen University (R07003 to JLZ).