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Stiffened tumor microenvironment enhances perineural invasion in breast cancer via integrin signaling

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Abstract

Background

Accumulating studies have shown that tumors are regulated by nerves, and there is abundant nerve infiltration in the tumor microenvironment. Many solid tumors including breast cancer (BRCA) have different degrees of perineural invasion (PNI), which is closely related to the tumor occurrence and progression. However, the regulatory mechanism of PNI in BRCA remains largely unexplored.

Methods

PNI-related molecular events are analyzed by the RNAseq data of BRCA samples deposited in The Cancer Genome Atlas (TCGA) database. Extracellular matrix (ECM) components within the tumor microenvironment are analyzed by immunohistochemical staining of α-SMA, Sirius red staining, and Masson trichrome staining. Soft and stiff matrix gels, living cell imaging, and dorsal root ganglion (DRG) coculture assay are used to monitor cancer cell invasiveness towards nerves. Western blotting, qRT-PCR, enzyme-linked immunosorbent assay combined with neutralizing antibody and small molecular inhibitors are employed to decode molecular mechanisms.

Results

Comparative analysis that the ECM was significantly associated with PNI status in the TCGA cohort. BRCA samples with higher α-SMA activity, fibrillar collagen, and collagen content had higher frequency of PNI. Compared with soft matrix, BRCA cells cultured in stiff matrix not only displayed higher cell invasiveness to DRG neurons but also had significant neurotrophic effects. Mechanistically, integrin β1 was identified as a functional receptor to the influence of stiff matrix on BRCA cells. Moreover, stiffened matrix-induced activation of integrin β1 transduces FAK-YAP signal cascade, which enhances cancer invasiveness and the neurotrophic effects. In clinical setting, PNI-positive BRCA samples had higher expression of ITGB1, phosphorylated FAK, YAP, and NGF compared with PNI-negative BRCA samples.

Conclusions

Our findings suggest that stiff matrix induces expression of pro-metastatic and neurotrophic genes through integrin β1-FAK-YAP signals, which finally facilitates PNI in BRCA. Thus, our study provides a new mechanism for PNI in BRCA and highlights nerve-based tumor treatment strategies.

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Data availability

All data that support the findings of this study are available from the corresponding authors upon reasonable request.

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Funding

This research was funded by National Natural Science Youth Foundation of China (Grant Nos. 82203558) and Natural Science Foundation of Jilin Provincial Department of science and technology (20210401057YY).

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Authors and Affiliations

  1. Departments of Breast Surgery, General Surgery Center, The First Hospital of Jilin University, No.71, Xinmin Street, Changchun City, Jilin Province, 130021, People’s Republic of China

    Bing Han, Xin Guan, Mingyue Ma, Baoling Liang, Yutong Liu, Ye Du & Dong Song

  2. Fenghuangyudu subdistrict Longquan community, Fenggang county, Zunyi, Guizhou, 564200, People’s Republic of China

    Linglie Ren

  3. State Key Laboratory of Systems Medicine for Cancer, Shanghai Cancer Institute, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200240, People’s Republic of China

    Shu-Heng Jiang

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  1. Bing Han
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  2. Xin Guan
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Contributions

BH, XG, M-YM, B-LL, LL-R, and Y-TL performed clinical analysis, in vitro cell experiments, manuscript preparation, and statistical analysis; YD contributed to bioinformatics analysis; SD and S-HJ, conceived, designed, supervised, analyzed and interpreted the study and provided critical review.

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Correspondence to Dong Song.

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This study was approved by the ethics committee of The First Hospital of Jilin University.

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Han, B., Guan, X., Ma, M. et al. Stiffened tumor microenvironment enhances perineural invasion in breast cancer via integrin signaling. Cell Oncol. (2023). https://doi.org/10.1007/s13402-023-00901-x

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