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NNMT switches the proangiogenic phenotype of cancer-associated fibroblasts via epigenetically regulating ETS2/VEGFA axis

Oncogene volume 43, pages 2647–2660 (2024)Cite this article

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Abstract

Cancer-associated fibroblasts (CAFs) are known to promote angiogenesis in oral squamous cell carcinoma (OSCC). However, the epigenetic mechanisms through which CAFs facilitate angiogenesis within the tumor microenvironment are still poorly characterized. Nicotinamide N’-methyltransferase (NNMT), a member of the N-methyltransferase family, was found to be a key molecule in the activation of CAFs. This study shows that NNMT in fibroblasts contributes to angiogenesis and tumor growth through an epigenetic reprogramming-ETS2-VEGFA signaling axis in OSCC. Single-cell RNA Sequencing (scRNA-seq) analysis suggests that NNMT is mainly highly expressed in fibroblasts of head and neck squamous cell carcinoma (HNSCC). Moreover, analysis of the TCGA database and multiple immunohistochemical staining of clinical samples also identified a positive correlation between NNMT and tumor angiogenesis. This research further employed an assembled organoid model and a fibroblast-endothelial cell co-culture model to authenticate the proangiogenic ability of NNMT. At the molecular level, high expression of NNMT in CAFs was found to promote ETS2 expression by regulating H3K27 methylation level through mediating methylation deposition. Furthermore, ETS2 was verified to be an activating transcription factor of VEGFA in this study. Collectively, our findings delineate an epigenetic molecular regulatory network of angiogenesis and provide a theoretical basis for exploring new targets and clinical strategy in OSCC.

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Fig. 1: NNMT is mainly highly expressed in fibroblasts and is closely associated with tumor angiogenesis.
Fig. 2: NNMT promotes the proangiogenic ability of fibroblasts via regulating VEGFA expression.
Fig. 3: Stromal NNMT accelerates angiogenesis in vitro assembled organoid and in vivo.
Fig. 4: ETS2 is a NNMT-regulated transcription factor mediating VEGFA upregulation in CAFs.
Fig. 5: NNMT induces ETS2 expression by inhibiting the tri-methylation of H3K27.
Fig. 6: NNMT-ETS2-VEGFA signal axis orchestrates the proangiogenic properties of CAFs.
Fig. 7: Schematic diagram of the mechanisms of NNMT mediated methyl sink-ETS2-VEGFA signaling axis in promoting angiogenesis of OSCC.

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Data availability

The data generated in this study are available within the article and its supplementary data files. The data that support the findings of this study are available in Figshare (https://doi.org/10.6084/m9.figshare.25230431).

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Acknowledgements

We would like to thank Dr. Xiazhou Fu (Wuhan University) for his expert guidance. This study was supported by grants from National Key R&D Program of China (No. 2022YFC2504200), the National Nature Science Foundation of China (No. 82273306; No.82373093).

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Authors and Affiliations

  1. State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, Key Laboratory of Oral Biomedicine Ministry of Education, Hubei Key Laboratory of Stomatology, School & Hospital of Stomatology, Wuhan University, Wuhan, China

    Xinmiao Wang, Hui Zhao, Xinyue Luo, Yang Chen, Congyu Shi, Yifan Wang, Junqiang Bai, Zhe Shao & Zhengjun Shang

  2. Department of Oral and Maxillofacial-Head and Neck Oncology, School of Stomatology-Hospital of Stomatology, Wuhan University, Wuhan, China

    Hui Zhao, Zhe Shao & Zhengjun Shang

  3. Department of Oral and Maxillofacial Surgery, School & Hospital of Stomatology, Wuhan University, Wuhan, China

    Yang Chen

  4. Day Surgery Center, School and Hospital of Stomatology, Wuhan University, Wuhan, China

    Zhe Shao

  5. Taikang Center for Life and Medical Sciences of Wuhan University, Wuhan, China

    Zhengjun Shang

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Contributions

Conceptualization, WXM, and ZH; Literature Search, WXM and LXY; Data Collection, WXM and SCY; Formal Analysis, WXM; Validation, WXM, LXY, SCY, WYF, and BJQ; Investigation and Visualization, WXM; Methodology, WXM and LXY; Writing-original draft, WXM; Project administration, SZ and SZJ; Writing-review & editing, LXY, CY, SZ and SZJ; Supervision, SZJ. All authors read and approved the final version of the manuscript.

Corresponding authors

Correspondence to Zhe Shao or Zhengjun Shang.

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Human material and human data involved in this research is in accordance with the Declaration of Helsinki. All animal studies were approved by the Ethics Committee of the School and Hospital of Stomatology, Wuhan University (Approval Number: S07921100A). Informed consent was obtained from all participating patients. This research did not involve identifiable images of human participants.

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NNMT switches the proangiogenic phenotype of cancer-associated fibroblasts via epigenetically regulating ETS2/VEGFA axis

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Wang, X., Zhao, H., Luo, X. et al. NNMT switches the proangiogenic phenotype of cancer-associated fibroblasts via epigenetically regulating ETS2/VEGFA axis. Oncogene 43, 2647–2660 (2024). https://doi.org/10.1038/s41388-024-03112-2

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