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Published on: November 28, 2015
METTL14/YTHDF2 m6A Axis Protects Against M2 Macrophage Polarization in Endometriosis by Regulating KLF4 Stability
Li Li1, Xiaoyan Wang2, You Wu2
1Departnent of Reproductive Medicine, Affiliated Hospital of Zunyi Medical University, No. 149 Dalian Road, Zunyi, Guizhou Province, 563000, China. yi111_157@163.com.
Abstract:
Endometriosis (EMs) is a chronic inflammatory disorder featured by infertility and pain. The role of N6-methyladenosine (m6A) in EMs has been evidenced. This study investigated the molecular mechanism of METTL14-m6A-KLF4 modulating macrophage polarization in EMs. RT-qPCR assay was conducted to test METTL14 levels in tissues and cells and the relative mRNA levels of M2 (Arg-1, Fizz1) and M1 (iNOS, IL-1β) factors in the supernatant after co-culture of macrophages with normal endometrial stromal cells (nESCs) or ectopic endometrial stromal cells (eESCs). CD206 and CD86 expression, as well as Arg-1, IL-10, and IL-4 levels, were assessed. Meanwhile, the relationship between METTL14 and the m6A modification of KLF4 was analyzed. Additionally, the effect of KLF4-activated M2 macrophages on in vitro ESC progression was observed. Cellular and tissue METTL14 was under-expressed in EMs. METTL14 expression might be related to macrophage M2 polarization. Co-culture of eESCs overexpressing METTL14 and macrophages downregulated Arg-1, Fizz1, CD206, IL-10, and IL-4 levels. Mechanistically, METTL14 could mediate KLF4 m6A modification through the m6A reading protein YTHDF2. KLF4 overexpression could nullify METTL14 re-expression-repressed M2 macrophage polarization. In addition, KLF4-activated M2 macrophages accelerated the proliferation and migration of ESCs in vitro. METTL14-m6A-KLF4 regulated macrophage polarization in EMs.
Insights
The METTL14-m6A-KLF4 pathway influences macrophage polarization in endometriosis (EMs). This study reveals METTL14
Area of Science:
- Reproductive Biology
- Immunology
- Molecular Biology
Background:
- Endometriosis (EMs) is a chronic inflammatory condition linked to infertility and pain.
- N6-methyladenosine (m6A) modification plays a role in EMs pathogenesis.
- Understanding molecular mechanisms, like METTL14-m6A-KLF4 interactions, is crucial for EMs research.
Purpose of the Study:
- To investigate the molecular mechanism of how METTL14-m6A-KLF4 regulates macrophage polarization in endometriosis.
- To explore the impact of METTL14 and KLF4 on macrophage phenotypes (M1/M2) in the context of EMs.
- To assess the effect of KLF4-activated M2 macrophages on endometrial stromal cell (ESC) progression.
Main Methods:
- RT-qPCR to quantify METTL14 mRNA levels in tissues and cells.
- Co-culture assays of macrophages with normal (nESCs) and ectopic (eESCs) endometrial stromal cells.
- Assessment of M1/M2 macrophage markers (e.g., iNOS, IL-1β, Arg-1, IL-4, CD206, CD86) and KLF4 m6A modification.
Main Results:
- METTL14 was underexpressed in EMs tissues and cells, correlating with M2 macrophage polarization.
- Overexpression of METTL14 in eESCs co-cultured with macrophages downregulated M2 markers.
- METTL14 mediated KLF4 m6A modification via YTHDF2, and KLF4 overexpression counteracted METTL14's effect on M2 polarization.
- KLF4-activated M2 macrophages promoted ESC proliferation and migration in vitro.
Conclusions:
- The METTL14-m6A-KLF4 axis is a key regulator of macrophage polarization in endometriosis.
- Dysregulation of this pathway may contribute to EMs development and progression.
- Targeting the METTL14-m6A-KLF4 pathway could offer therapeutic strategies for endometriosis.
