Role of M1 macrophages in diabetic foot ulcers and related immune regulatory mechanisms

Yao Li; Xiaoyan Li; Shuai Ju; Wenqiang Li; Siyuan Zhou; Guili Wang; Yunmin Cai; Zhihui Dong

doi:10.3389/fphar.2022.1098041

Role of M1 macrophages in diabetic foot ulcers and related immune regulatory mechanisms

Front Pharmacol. 2023 Jan 9:13:1098041. doi: 10.3389/fphar.2022.1098041. eCollection 2022.

Authors

Yao Li^{1

2}, Xiaoyan Li^{1

2}, Shuai Ju^{1

2}, Wenqiang Li^{1

2}, Siyuan Zhou^{1

2

3}, Guili Wang^{1

2

3}, Yunmin Cai^{1

2}, Zhihui Dong^{1

2

4}

Affiliations

¹ Jinshan Hospital, Fudan University, Shanghai, China.
² Zhongshan Diabetic foot Multidisciplinary Diagnosis and Treatment Center and Jinshan Operation Center, Shanghai, China.
³ Shanghai Medical College, Fudan University, Shanghai, China.
⁴ Zhongshan Hospital, Fudan University, Shanghai, China.

Abstract

Objectives: Diabetes foot ulcers (DFUs) are characterized by immune infiltration of M1 macrophages observed in foot skin, in which immune-associated genes (IRGs) play a prominent role. The precise expression of IRGs as well as any possible regulatory mechanisms that could be present in DFUs is yet unknown. Methods: The sequencing data of single-cell RNA (scRNA) in the foot skin of patients with DFUs were analyzed, screening out the cluster marker genes of foot skin obtained from the ImmPort database. IRG activity was assessed with the AUCell software package. The IRGs of DFUs were explored by analyzing the batch sequencing dataset of DFU skin tissue. HumanTFDB was adopted to identify relevant regulatory transcription factors (TFs). The STRING dataset was used to build the main TF protein-protein interaction networks. WB and immunofluorescence methods were used to verify M1 macrophage-related immune regulators. Results: There were 16 clusters found: SMC1, fibro, t-lympho, he fibro, vasendo, baselkera, diffkera, SMC2, M1 macro, M2 macro, sweet/seba, B-Lympho, Melanio, lymphendo, plasma, and Schwann. M1 and M2 macrophages both had considerably higher AUC ratings than patients with DFUs compared to other sub-populations of cells. The proportion of M1 macrophages was the highest in the non-healing group. According to scRNA analysis and batch sequencing data by GO and KEGG, DEGs were enriched in immune response. Some 106 M1 macro-IRGs were finally identified and 25 transcription factors were revealed as associated with IRG expression. The PPI network indicated NFE2L2, REL, ETV6, MAF, and NF1B as central transcription factors. Conclusion: Based on the bio-informatics analysis of scRNA and high-throughput sequencing data, we concluded that M1 macrophages may serve as the influencing factor of DFUs' non-union. In addition, NFE2L2 could be involved in the regulation of IRG expression within M1 macrophages.

Keywords: M1 macrophages; diabetes foot ulcer; gene associated with immunity; immune infiltration; sequencing of single cell.

Grants and funding

The research was supported by the National Natural Science Foundation of China (General Program, No. 81970407) 2021 “Famous Clinical Doctor Training Project” of Fudan University (No. DGF828008/001/002), Research Project of Shanghai Jinshan District Health Commission (No. JSKJ-KTMS-2020–02), and Youth Research Initiation Fund of Jinshan Hospital Affiliated to Fudan University (No.JYQN-JC-202214).