TET3-overexpressing macrophages promote endometriosis
Haining Lv, Beibei Liu, Yangyang Dai, Feng Li, Stefania Bellone, Yuping Zhou, Ramanaiah Mamillapalli, Dejian Zhao, Muthukumaran Venkatachalapathy, Yali Hu, Gordon G. Carmichael, Da Li, Hugh S. Taylor, Yingqun Huang
Haining Lv, Beibei Liu, Yangyang Dai, Feng Li, Stefania Bellone, Yuping Zhou, Ramanaiah Mamillapalli, Dejian Zhao, Muthukumaran Venkatachalapathy, Yali Hu, Gordon G. Carmichael, Da Li, Hugh S. Taylor, Yingqun Huang
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Research Article Inflammation Reproductive biology

TET3-overexpressing macrophages promote endometriosis

Abstract

Endometriosis is a debilitating, chronic inflammatory disease affecting approximately 10% of reproductive-age women worldwide with no cure. While macrophages have been intrinsically linked to the pathophysiology of endometriosis, targeting them therapeutically has been extremely challenging due to their high heterogeneity and because these disease-associated macrophages (DAMs) can be either pathogenic or protective. Here, we report identification of pathogenic macrophages characterized by TET3 overexpression in human endometriosis lesions. We show that factors from the disease microenvironment upregulated TET3 expression, transforming macrophages into pathogenic DAMs. TET3 overexpression stimulated proinflammatory cytokine production via a feedback mechanism involving inhibition of let-7 miRNA expression. Remarkably, these cells relied on TET3 overexpression for survival and hence were vulnerable to TET3 knockdown. We demonstrated that Bobcat339, a synthetic cytosine derivative, triggered TET3 degradation in both human and mouse macrophages. This degradation was dependent on a von Hippel-Lindau (VHL) E3 ubiquitin ligase whose expression was also upregulated in TET3-overexpressing macrophages. Furthermore, depleting TET3-overexpressing macrophages either through myeloid-specific Tet3 ablation or using Bobcat339 strongly inhibited endometriosis progression in mice. Our results defined TET3-overexpressing macrophages as key pathogenic contributors to and attractive therapeutic targets for endometriosis. Our findings may also be applicable to other chronic inflammatory diseases where DAMs have important roles.

Authors

Haining Lv, Beibei Liu, Yangyang Dai, Feng Li, Stefania Bellone, Yuping Zhou, Ramanaiah Mamillapalli, Dejian Zhao, Muthukumaran Venkatachalapathy, Yali Hu, Gordon G. Carmichael, Da Li, Hugh S. Taylor, Yingqun Huang

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Figure 8

TET3 enhances IL-1β and IL-6 expression by decreasing let-7 miRNA levels.

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TET3 enhances IL-1β and IL-6 expression by decreasing let-7 miRNA levels...
(A) qRT-PCR of let-7a in RAW 264.7 cells transfected with NT siRNA or Tet3 siRNA. RNA was isolated at 24 hours after transfection. n = 3 technical replicates. (B) qRT-PCR of let-7a in PMs isolated from WT and Mye-Tet3–KO mice. n = 3 mice per genotype. (C) PMs were isolated from WT mice and treated with TGF-β1 at a final concentration of 30 ng/mL. After 48 hours, Veh or Bc was added at a final concentration of 10 μM and incubation carried out for 48 hours. RNAs were extracted and analyzed by qRT-PCR. n = 3 mice per group. (D) qRT-PCR of Lin28b mRNA in PMs isolated from WT and Mye-Tet3–KO mice. n = 3 mice per genotype. (E) qRT-PCR of Lin28b mRNA in PMs isolated from WT mice and treated as in C. n = 3 mice per group. (F) qRT-PCR of Il-1b and Il-6 mRNAs of PMs isolated from WT mice and transfected with control miRNA (miCon) or let-7a mimic and stimulated with 10 ng/mL LPS plus 20 ng/mL IFN-γ. RNAs were isolated after 6 hours of LPS/IFN-γ stimulation. n = 3 mice per group. (G) ELISA results of IL-1β (after 6 hours of LPS/IFN-γ stimulation) and IL-6 (after 10 hours of LPS/IFN-γ stimulation) of PMs treated as in F. n = 3 mice per group. (H) Relative let-7a miRNA levels in endometriosis lesions from WT and Mye-tet3–KO mice. n = 4 animals per genotype. (I) Relative let-7a miRNA levels in endometriosis lesions from Veh- or Bc-treated mice. n = 4 animals per group. All data are represented as mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001, 2-tailed Student’s t test.