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Schwann cell derived pleiotrophin stimulates fibroblast for proliferation and excessive collagen deposition in plexiform neurofibroma

Cancer Gene Therapy volume 31, pages 627–640 (2024)Cite this article

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Abstract

Neurofibromatosis type 1 associated plexiform neurofibroma (pNF) is characterized by abundant fibroblasts and dense collagen, yet the intricate interactions between tumor-origin cells (Schwann cells) and neurofibroma-associated fibroblasts (NFAFs) remain elusive. Employing single-cell RNA sequencing on human pNF samples, we generated a comprehensive transcriptomics dataset and conducted cell-cell communication analysis to unravel the molecular dynamics between Schwann cells and NFAFs. Our focus centered on the pleiotrophin (PTN)/nucleolin (NCL) axis as a pivotal ligand-receptor pair orchestrating this interaction. Validation of PTN involvement was affirmed through coculture models and recombinant protein experiments. Functional and mechanistic investigations, employing assays such as CCK8, EdU, Western Blot, ELISA, Hydroxyproline Assay, and Human phospho-kinase array, provided critical insights. We employed siRNA or inhibitors to intercept the PTN/NCL/proline-rich Akt substrate of 40 kDa (PRAS40) axis, validating the associated molecular mechanism. Our analysis highlighted a subset of Schwann cells closely linked to collagen deposition, underscoring their significance in pNF development. The PTN/NCL axis emerged as a key mediator of the Schwann cell-NFAF interaction. Furthermore, our study demonstrated that elevated PTN levels enhanced NFAF proliferation and collagen synthesis, either independently or synergistically with TGF-β1 in vitro. Activation of the downstream molecule PRAS40 was noted in NFAFs upon PTN treatment. Crucially, by targeting NCL and PRAS40, we successfully reversed collagen synthesis within NFAFs. In conclusion, our findings unveil the pivotal role of the PTN/NCL/PRAS40 axis in driving pNF development by promoting NFAFs proliferation and function. Targeting this pathway emerges as a potential therapeutic strategy for pNF. This study contributes novel insights into the molecular mechanisms governing pNF pathogenesis.

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Fig. 1: Transcriptomic identification of major cell types that comprise the NF1 related pNF.
Fig. 2: Molecular profile of Schwann Cell subtype heterogeneity of pNF.
Fig. 3: Molecular profiles of Fibroblast subtype heterogeneity of pNF.
Fig. 4: Ligand–receptor analysis to assess cell interaction between Schwann cells and Fibroblasts.
Fig. 5: Characterization of the PTN in human clinical samples (skin (normal skin), keloid, never (normal peripheral nerve), pNF) by immunohistochemistry.
Fig. 6: PTN/NCL expression and cellular localization in Schwann cells and NFAFs.
Fig. 7: Coculturing with ipNF95.11bC, sNF96.2 promoted proliferation and regulated collagen synthesis of NFAFs.
Fig. 8: Cell signaling analysis for NFAFs in response to PTN.

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Data availability

The datasets used and/or analysed during the current study available from the corresponding author on reasonable request.

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Funding

This work was supported by the National Natural Science Foundation of China [No. 81870780, No. 81902746]. Shanghai Municipal Health Commission Program [No. 20194Y0017].

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Author notes

  1. These authors contributed equally: Zhuowei Tian, Zhong Du, Guo Bai, Qiyu Gong.

Authors and Affiliations

  1. Department of Oral Maxillofacial-Head and Neck Oncology, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology, Shanghai, China

    Zhuowei Tian, Zhong Du, Guo Bai, Yuanhe You, Guisong Xu, Meng Xiao, Yanan Wang & Yue He

  2. Department of Oral Maxillofacial-Head and Neck Oncology, Fengcheng Hospital, Shanghai, China

    Zhuowei Tian & Meng Xiao

  3. Institute of Immunology, Faculty of Basic Medicine, Shanghai Jiao Tong University School of Medicine, Shanghai, China

    Qiyu Gong

  4. Department of Oral Maxillofacial Surgery, Shanghai Stomatological Hospital & School of Stomatology, Fudan University, Shanghai, China

    Jialiang Liu

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Contributions

ZT and ZD: Data curation; Biological experiment; Original draft preparation. GB and QG: Software; Visualization; Original draft preparation. YY, GX, JL: Data curation; Formal analysis. MX, YW, YH: Supervision; Conceptualization and Review & editing the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Meng Xiao, Yanan Wang or Yue He.

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This study was approved by the Medical Committee of Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China (SH9H-2021-TK253-1). Written consents were also obtained prior to the inception of this study. Written informed consent was received for the use of the photographs and that the record of informed consent has been retained.

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Tian, Z., Du, Z., Bai, G. et al. Schwann cell derived pleiotrophin stimulates fibroblast for proliferation and excessive collagen deposition in plexiform neurofibroma. Cancer Gene Ther 31, 627–640 (2024). https://doi.org/10.1038/s41417-024-00727-1

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