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The development of in vitro organotypic 3D vulvar models to study tumor-stroma interaction and drug efficacy

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Abstract

Background

Vulvar squamous cell carcinoma (VSCC) is a rare disease with a poor prognosis. To date, there’s no proper in vitro modeling system for VSCC to study its pathogenesis or for drug evaluation.

Methods

We established healthy vulvar (HV)- and VSCC-like 3D full thickness models (FTMs) to observe the tumor-stroma interaction and their applicability for chemotherapeutic efficacy examination. VSCC-FTMs were developed by seeding VSCC tumor cell lines (A431 and HTB117) onto dermal matrices harboring two NF subtypes namely papillary fibroblasts (PFs) and reticular fibroblasts (RFs), or cancer-associated fibroblasts (CAFs) while HV-FTMs were constructed with primary keratinocytes and fibroblasts isolated from HV tissues.

Results

HV-FTMs highly resembled HV tissues in terms of epidermal morphogenesis, basement membrane formation and collagen deposition. When the dermal compartment shifted from PFs to RFs or CAFs in VSCC-FTMs, tumor cells demonstrated more proliferation, EMT induction and stemness. In contrast to PFs, RFs started to lose their phenotype and express robust CAF-markers α-SMA and COL11A1 under tumor cell signaling induction, indicating a favored ‘RF-to-CAF’ transition in VSCC tumor microenvironment (TME). Additionally, chemotherapeutic treatment with carboplatin and paclitaxel resulted in a significant reduction in tumor-load and invasion in VSCC-FTMs.

Conclusion

We successfully developed in vitro 3D vulvar models mimicking both healthy and tumorous conditions which serve as a promising tool for vulvar drug screening programs. Moreover, healthy fibroblasts demonstrate heterogeneity in terms of CAF-activation in VSCC TME which brings insights in the future development of novel CAF-based therapeutic strategies in VSCC.

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Data availability

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

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Acknowledgements

The authors would like to thank the Roosevelt Clinics and the Dermatology department of the LUMC for providing fresh vulvar tissues. The authors thank the donors of the Bontiusstichting (project 8222–32146) of the Leiden University Medical Center for their financial contribution. The authors would also like to thank China Scholarship Council (CSC) for supporting the stay of Shidi Wu in the Netherlands. Graphic abstract, Figs. 1a and 2a were created with BioRender.com.

Funding

This study was partially funded by the Bontiusstichting (project 8222–32146).

Author information

Authors and Affiliations

  1. Department of Dermatology, Leiden University Medical Center, Leiden, 2333 ZA, The Netherlands

    Shidi Wu, Marion H. Rietveld, Robert Rissmann, Maarten H. Vermeer & Abdoelwaheb El Ghalbzouri

  2. Center for Human Drug Research, Leiden, 2333 CL, The Netherlands

    Bertine W. Huisman, Robert Rissmann & Mariette I. E. van Poelgeest

  3. Department of Gynecology, Leiden University Medical Center, Leiden, 2333 ZA, The Netherlands

    Bertine W. Huisman & Mariette I. E. van Poelgeest

  4. Leiden Academic Center for Drug Research, Leiden University, Leiden, 2333 CC, The Netherlands

    Robert Rissmann

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Contributions

Conceptualization: S.W., B.H, M.P. and A.E.G. Investigation and formal analysis: S.W., B.H and M.R. Writing-original draft preparation: S.W. and B.H. Writing-review and editing: M.P., A.E.G., M.V., and R.R. Supervision: M.P., A.E.G, M.V., and R.R. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Abdoelwaheb El Ghalbzouri.

Ethics declarations

Ethics approval

Declaration of Helsinki principles were followed during the obtainment of primary cells from human skin originating from surplus breast tissue, vulvar labial tissue and cutaneous tumor biopsies. All subjects remained anonymous. Experiments were conducted in accordance with article 7:467 of the Dutch Law on Medical Treatment Agreement and the Code for Proper Use of Human Tissue of the Dutch Federation of Biomedical Scientific Societies (https://www.federa.org). According to this national legislation, surplus tissue can be used for scientific research purposes when no written objection is made by the informed donor. Therefore, additional approval of an ethics committee regarding the scientific use of surplus tissue was not obligatory.

Consent for publication

For the manuscript solitary tissue of anonymous donors was used, no individual person’s data was included.

Competing interests

The authors declare no competing interests.

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Shidi Wu and Bertine W. Huisman contributed equally to this work, shared first authorship.

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Wu, S., Huisman, B.W., Rietveld, M.H. et al. The development of in vitro organotypic 3D vulvar models to study tumor-stroma interaction and drug efficacy. Cell Oncol. (2023). https://doi.org/10.1007/s13402-023-00902-w

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