Alteration of the m⁶A Methylation Landscape in a Mouse Model of Scleroderma

Abstract

Aim: To explore the N⁶-methyladenosine (m⁶A) methylation of mRNAs and its roles in a mouse model of scleroderma. Materials & methods: To evaluate whether the mouse model of scleroderma could meet the experimental requirements, we examined skin tissue specimens by pathological staining and identified the related indicators by quantitative PCR  (qPCR). m⁶A-tagged mRNAs were identified via m⁶A epitranscriptomic microarray, and m⁶A-RNA-immunoprecipitation qPCR and qPCR were performed to confirm microarray data. Results: There were differences in m⁶A methylation among 843 mRNAs. Further, there were significant differences among Hras, Saa1, Ccl3, Ccl9 and Il1b in terms of methylation and expression. Conclusion: The m⁶A methylation spectrum in a mouse model of scleroderma may explain the occurrence of scleroderma.

Lay abstract

Scleroderma is an autoimmune disease with an unknown etiology. At present there is no effective treatment, and the prognosis is poor. Although m⁶A methylation is the most common RNA modification and affects cell biological function by influencing mRNA expression levels, the role of m⁶A methylation in scleroderma is unknown. Based on a mouse model of scleroderma, we screened and validated mRNAs related to m⁶A regulation, including Hras, Lama3, Stat3, Tnc, Ccl3, Ccl9, Saa1 and Il1b. While the results are encouraging, they are only preliminary. We plan to explore the altered pattern of m⁶A methylation in skin specimens from patients with scleroderma. This study is important for developing biomarkers for diagnosis and treatment of scleroderma in the future.

Keywords::

• bleomycin
• methylation
• mRNA
• N⁶-methyladenosine
• scleroderma

Supplementary data

To view the supplementary data that accompany this paper please visit the journal website at: www.tandfonline.com/doi/suppl/10.2217/epi-2021-0369

Financial & competing interests disclosure

This work is supported by the grants from the National Natural Science Foundation of China, which is a subsidiary of the Ministry of Science and Technology, PRC. The General Program number is 81874240. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

Writing assistance was provided by Enago and was funded by the National Natural Science Foundation of China (program no. 81874240).

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all animal experimental investigations, and all experiments were approved and performed following the guidelines of the Tongji University Ethic committee of China (no. TJ-HB-LAC-2020-83).

Additional information

Funding

This work is supported by the grants from the National Natural Science Foundation of China, which is a subsidiary of the Ministry of Science and Technology, PRC. The General Program number is 81874240. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. Writing assistance was provided by Enago and was funded by the National Natural Science Foundation of China (program no. 81874240).