Genome-wide screening of m6A profiling of cutaneous wound healing in diabetic mice
Junjie Shen1, Hua Chen2, Jiezhi Dai3
1Department of Orthopedic Surgery, Shanghai Sixth People's Hospital, JiaoTong University, Shanghai, China.
Objective:
Impaired wound healing in diabetes mellitus (DM) is a major health burden on patients, their families, and society. The present study aimed to systematically profile the m6A modification landscape in cutaneous wounds in a diabetic mouse model.
Approach:
Diabetes was induced in mice through a single intraperitoneal injection of streptozotocin (STZ); a single intraperitoneal injection of PBS was made in control mice for comparisons. Both groups then received an 8-mm diameter, full-thickness dorsal body wound with a biopsy punch. Five days after wound surgery, western blot analysis of harvested wound tissues from both groups was used to assess the expression of m6A-related enzymes. Genome-wide profiling of m6A-tagged transcripts was performed through MeRIP-seq and RNA-seq.
Results:
ALKBH5, an m6A eraser, was significantly upregulated, while METTL3, METTL14, and WTAP, m6A writers, were markedly downregulated in the diabetic wounds. Additionally, a total of 1335 m6A peaks were differentially expressed in MeRIP-seq and RNA-seq analyses, with 558 upregulated and 777 downregulated peaks. Finally, there was hypomethylated and hypermethylated differentiation at the gene and transcript levels.
Innovation:
The present study was the first to reveal the m6A landscape in diabetic wounds in an animal model.
Conclusion:
This study, by deeply analyzing the role of m6A modifications in diabetic wound healing, provides new insights and understanding into the molecular mechanisms of diabetic wound healing. Future research could further explore how m6A modifications regulate the wound healing process, thereby offering new potential targets for the treatment of diabetic wounds.
Insights
This study profiled RNA modifications in diabetic mouse wounds, revealing altered m6A enzyme levels and gene expression. These findings offer new molecular insights into impaired diabetic wound healing.
Area of Science:
- Biochemistry
- Molecular Biology
- Wound Healing Research
Background:
- Diabetes mellitus (DM) significantly impairs wound healing, posing a major health challenge.
- Understanding the molecular mechanisms underlying impaired diabetic wound healing is crucial for developing effective treatments.
Purpose of the Study:
- To systematically profile the N6-methyladenosine (m6A) modification landscape in cutaneous wounds of a diabetic mouse model.
- To investigate the differential expression of m6A-related enzymes and transcripts in diabetic wounds.
Main Methods:
- Diabetes was induced in mice using streptozotocin (STZ).
- Full-thickness skin wounds were created and analyzed after five days.
- Western blot was used to assess m6A enzyme expression.
- MeRIP-seq and RNA-seq were employed for genome-wide m6A profiling.
Main Results:
- Key m6A 'eraser' enzyme ALKBH5 was upregulated, while 'writer' enzymes (METTL3, METTL14, WTAP) were downregulated in diabetic wounds.
- 1335 differentially expressed m6A peaks were identified (558 upregulated, 777 downregulated).
- Distinct patterns of hypomethylation and hypermethylation were observed at gene and transcript levels.
Conclusions:
- This study provides the first comprehensive m6A landscape analysis in a diabetic wound animal model.
- The findings offer novel insights into the molecular mechanisms of diabetic wound healing.
- m6A modifications represent potential therapeutic targets for improving diabetic wound healing.
