Department of Pathology, Shanghai Ninth People’s Hospital, affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China
Department of Dermatology and Dermatologic Surgery, Shanghai Ninth People’s Hospital, affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China
Department of Plastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital, affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China
Organ Regeneration X Lab, LiSheng East China Institute of Biotechnology, Peking University, Nantong 226299, China
§ Yanghua Shi and Jiping Liu contributed equally to this work.
• Bermatofibrosarcoma protuberans (DFSP) organoids used to test responses to imatinib and metformin
• Metformin inhibits the growth of DFSP organoids via immune signaling pathway
Graphical Abstract
Here, we developed patient-derived skin tumor organoids mimicking clinical tissues, showcasing diverse cell types and immune interactions. Single-cell sequencing identified 11 cell types, highlighting fidelity to in vivo counterparts. Bermatofibrosarcoma protuberans (DFSP) organoids revealed metformin's unique immune signaling modulation, aiding drug testing and mechanistic exploration.
Abstract
Surgery is the primary treatment for skin tumors, but it can result in scarring and distress for patients. Developing alternative therapeutic methods necessitates suitable in vitro models, which are currently limited in accurately representing the in vivo cell types and microenvironment of skin tumors. Here, we present a practical approach for creating patient-derived skin tumor organoids that effectively replicate the histological characteristics and mutational profiles observed in clinical tissues. Utilizing single-cell sequencing, we identified up to 11 distinct cell types within the organoid samples, encompassing various skin system cells and immune cells. Furthermore, we demonstrate the applicability of bermatofibrosarcoma protuberans (DFSP) organoids for assessing their responses to imatinib and metformin. Our findings reveal that metformin, in contrast to imatinib, can modulate the expression of downstream genes through immune signaling pathways. Our results underscore the ability of DFSP organoids to preserve key features of clinical tissues, including the presence of multiple cell types, especially immune cells. Importantly, our organoids provide a convenient approach for investigating the effects of drugs and elucidating underlying molecular mechanisms.
No abstract is available for this article. Click the button above to view the PDF directly.
Electronic Supplementary Material
Download File(s)
CO-2024-0001_ESM.pdf (690.8 KB)
References
[1]
Tay, S. S., Roediger, B., Tong, P. L., Tikoo, S., Weninger, W. The skin-resident immune network. Current Dermatology Reports, 2014, 3(1): 13–22. https://doi.org/10.1007/s13671-013-0063-9
Mestrallet, G., Rouas-Freiss, N., LeMaoult, J., Fortunel, N. O., Martin, M. T. Skin immunity and tolerance: Focus on epidermal keratinocytes expressing HLA-G. Frontiers in Immunology, 2021, 12: 772516. https://doi.org/10.3389/fimmu.2021.772516
Nestle, F. O., Di Meglio, P., Qin, J. Z., Nickoloff, B. J. Skin immune sentinels in health and disease. Nature Reviews Immunology, 2009, 9(10): 679–691. https://doi.org/10.1038/nri2622
Wang, T., Li, K., Xiao, S. X., Xia, Y. M. A plausible role for collectins in skin immune homeostasis. Frontiers in Immunology, 2021, 12: 594858. https://doi.org/10.3389/fimmu.2021.594858
Pasparakis, M., Haase, I., Nestle, F. O. Mechanisms regulating skin immunity and inflammation. Nature Reviews Immunology, 2014, 14(5): 289–301. https://doi.org/10.1038/nri3646
Johnson-Huang, L. M., McNutt, N. S., Krueger, J. G., Lowes, M. A. Cytokine-producing dendritic cells in the pathogenesis of inflammatory skin diseases. Journal of Clinical Immunology, 2009, 29(3): 247–256. https://doi.org/10.1007/s10875-009-9278-8
Turchin, I., Bourcier, M. The role of interleukins in the pathogenesis of dermatological immune-mediated diseases. Advances in Therapy, 2022, 39(10): 4474–4508. https://doi.org/10.1007/s12325-022-02241-y
Fetter, T., Niebel, D., Braegelmann, C., Wenzel, J. Skin-associated B cells in the pathogenesis of cutaneous autoimmune diseases—Implications for therapeutic approaches. Cells, 2020, 9(12): 2627. https://doi.org/10.3390/cells9122627
Xiong, D. D., Bordeaux, J. S. Incidence and survival outcomes of dermatofibrosarcoma protuberans from 2000 to 2020: A population-based retrospective cohort analysis. Dermatologic Surgery, 2023, 49(12): 1096–1103. https://doi.org/10.1097/dss.0000000000004018
Roh, M. R., Bae, B., Chung, K. Y. Mohs’ micrographic surgery for dermatofibrosarcoma protuberans. Clinical and Experimental Dermatology, 2010, 35(8): 849–852. https://doi.org/10.1111/j.1365-2230.2010.03819.x
Doufekas, K., Duncan, T. J., Williamson, K. M., Varma, S., Nunns, D. Mohs micrographic surgery for dermatofibrosarcoma protuberans of the vulva. Obstetrics and Gynecology International, 2009, 2009: 547672. https://doi.org/10.1155/2009/547672
Zwald, F. O. Underuse of mohs micrographic surgery for the treatment of dermatofibrosarcoma protuberans. Archives of Dermatology, 2012, 148(9): 1064. https://doi.org/10.1001/archdermatol.2012.2685
Zeitouni, N., Cavanaugh, K., DuPont, J. Dermatofibrosarcoma protuberans: An update and review. Current Dermatology Reports, 2015, 4(4): 195–204. https://doi.org/10.1007/s13671-015-0120-7
Oyama, R., Kito, F., Qiao, Z. W., Sakumoto, M., Shiozawa, K., Toki, S., Yoshida, A., Kawai, A., Kondo, T. Establishment of novel patient-derived models of dermatofibrosarcoma protuberans: Two cell lines, NCC-DFSP1-C1 and NCC-DFSP2-C1. In Vitro Cellular & Developmental Biology - Animal, 2019, 55(1): 62–73. https://doi.org/10.1007/s11626-018-0305-z
Tang, X. Y., Wu, S., Wang, D., Chu, C., Hong, Y., Tao, M., Hu, H., Xu, M., Guo, X., Liu, Y. Human organoids in basic research and clinical applications. Signal Transduction and Targeted Therapy, 2022, 7: 168. https://doi.org/10.1038/s41392-022-01024-9
Xu, H. X., Jiao, D. C., Liu, A. G., Wu, K. M. Tumor organoids: Applications in cancer modeling and potentials in precision medicine. Journal of Hematology & Oncology, 2022, 15(1): 58. https://doi.org/10.1186/s13045-022-01278-4
Zhou, Z., Cong, L., Cong, X. Patient-derived organoids in precision medicine: Drug screening, organoid-on-a-chip and living organoid biobank. Frontiers in Oncology, 2021, 11: 762184. https://doi.org/10.3389/fonc.2021.762184
Forsythe, S. D., Sivakumar, H., Erali, R. A., Wajih, N., Li, W. C., Shen, P., Levine, E. A., Miller, K. E., Skardal, A., Votanopoulos, K. I. Patient-specific sarcoma organoids for personalized translational research: Unification of the operating room with rare cancer research and clinical implications. Annals of Surgical Oncology, 2022, 29(12): 7354–7367. https://doi.org/10.1245/s10434-022-12086-y
Lei, M. X., Schumacher, L. J., Lai, Y. C., Juan, W. T., Yeh, C. Y., Wu, P., Jiang, T. X., Baker, R. E., Widelitz, R. B., Yang, L. et al. Self-organization process in newborn skin organoid formation inspires strategy to restore hair regeneration of adult cells. Proceedings of the National Academy of Sciences of the United States of America, 2017, 114(34): E7101–E7110. https://doi.org/10.1073/pnas.1700475114
Hong, Z. X., Zhu, S. T., Li, H., Luo, J. Z., Yang, Y., An, Y., Wang, X., Wang, K. Bioengineered skin organoids: From development to applications. Military Medical Research, 2023, 10(1): 40. https://doi.org/10.1186/s40779-023-00475-7
Navarrete-Dechent, C., Mori, S., Barker, C. A., Dickson, M. A., Nehal, K. S. Imatinib treatment for locally advanced or metastatic dermatofibrosarcoma protuberans: A systematic review. JAMA Dermatology, 2019, 155(3): 361–369. https://doi.org/10.1001/jamadermatol.2018.4940
Johnson-Jahangir, H., Sherman, W., Ratner, D. Using imatinib as neoadjuvant therapy in dermatofibrosarcoma protuberans: Potential pluses and minuses. Journal of the National Comprehensive Cancer Network, 2010, 8(8): 881–885. https://doi.org/10.6004/jnccn.2010.0065
Rutkowski, P., Wozniak, A., Switaj, T. Advances in molecular characterization and targeted therapy in dermatofibrosarcoma protuberans. Sarcoma, 2011, 2011: 959132. https://doi.org/10.1155/2011/959132
Deng, C. C., Hu, Y. F., Zhu, D. H., Cheng, Q., Gu, J. J., Feng, Q. L., Zhang, L. X., Xu, Y. P., Wang, D., Rong, Z. L. et al. Single-cell RNA-seq reveals fibroblast heterogeneity and increased mesenchymal fibroblasts in human fibrotic skin diseases. Nature Communications, 2021, 12: 3709. https://doi.org/10.1038/s41467-021-24110-y
Ge, L. L., Wang, Z. C., Wei, C. J., Huang, J. X., Liu, J., Gu, Y. H., Wang, W., Li, Q. F. Unraveling intratumoral complexity in metastatic dermatofibrosarcoma protuberans through single-cell RNA sequencing analysis. Cancer Immunology, Immunotherapy, 2023, 72(12): 4415–4429. https://doi.org/10.1007/s00262-023-03577-2
Kemper, K., Krijgsman, O., Cornelissen-Steijger, P., Shahrabi, A., Weeber, F., Song, J. Y., Kuilman, T., Vis, D. J., Wessels, L. F., Voest, E. E. et al. Intra- and inter-tumor heterogeneity in a vemurafenib-resistant melanoma patient and derived xenografts. EMBO Molecular Medicine, 2015, 7(9): 1104–1118. https://doi.org/10.15252/emmm.201404914
Kerkour, T., Zhou, C., Hollestein, L., Mooyaart, A. Genetic concordance in primary cutaneous melanoma and matched metastasis: A systematic review and meta-analysis. International Journal of Molecular Sciences, 2023, 24(22): 16281. https://doi.org/10.3390/ijms242216281
Skuli, S. J., Alomari, S., Gaitsch, H., Bakayoko, A., Skuli, N., Tyler, B. Metformin and cancer, an ambiguanidous relationship. Phamaceuticals, 2022, 15(5): 626. https://doi.org/10.3390/ph15050626
Hua, Y., Zheng, Y., Yao, Y. R., Jia, R. B., Ge, S. F., Zhuang, A. Metformin and cancer hallmarks: Shedding new lights on therapeutic repurposing. Journal of Translational Medicine, 2023, 21(1): 403. https://doi.org/10.1186/s12967-023-04263-8
Jacob, F., Salinas, R. D., Zhang, D. Y., Nguyen, P. T. T., Schnoll, J. G., Wong, S. Z. H., Thokala, R., Sheikh, S., Saxena, D., Prokop, S. et al. A patient-derived glioblastoma organoid model and biobank recapitulates inter- and intra-tumoral heterogeneity. Cell, 2020, 180(1): 188–204.e22. https://doi.org/10.1016/j.cell.2019.11.036
LeBlanc, V. G., Trinh, D. L., Aslanpour, S., Hughes, M., Livingstone, D., Jin, D., Ahn, B. Y., Blough, M. D., Cairncross, J. G., Chan, J. A. et al. Single-cell landscapes of primary glioblastomas and matched explants and cell lines show variable retention of inter- and intratumor heterogeneity. Cancer Cell, 2022, 40(4): 379–392.e9. https://doi.org/10.1016/j.ccell.2022.02.016
Choe, M. S., Kim, S. J., Oh, S. T., Bae, C. M., Choi, W. Y., Baek, K. M., Kim, J. S., Lee, M. Y. A simple method to improve the quality and yield of human pluripotent stem cell-derived cerebral organoids. Heliyon, 2021, 7(6): e07350. https://doi.org/10.1016/j.heliyon.2021.e07350
Hu, Y. W., Sui, X. Z., Song, F., Li, Y. Q., Li, K. Y., Chen, Z. Y., Yang, F., Chen, X. Y., Zhang, Y. H., Wang, X. N. et al. Lung cancer organoids analyzed on microwell arrays predict drug responses of patients within a week. Nature Communications, 2021, 12: 2581. https://doi.org/10.1038/s41467-021-22676-1
Shankaran, A., Prasad, K., Chaudhari, S., Brand, A., Satyamoorthy, K. Advances in development and application of human organoids. 3 Biotech, 2021, 11(6): 257. https://doi.org/10.1007/s13205-021-02815-7
Calà, G., Sina, B., De Coppi, P., Giobbe, G. G., Gerli, M. F. M. Primary human organoids models: Current progress and key milestones. Frontiers in Bioengineering and Biotechnology, 2023, 11: 1058970. https://doi.org/10.3389/fbioe.2023.1058970
Suarez-Martinez, E., Suazo-Sanchez, I., Celis-Romero, M., Carnero, A. 3D and organoid culture in research: Physiology, hereditary genetic diseases and cancer. Cell & Bioscience, 2022 , 12(1): 39. https://doi.org/10.1186/s13578-022-00775-w
Cruz-Gil, S., Sánchez-Martínez, R., Wagner-Reguero, S., Stange, D., Schölch, S., Pape, K., Ramírez de Molina, A. A more physiological approach to lipid metabolism alterations in cancer: CRC-like organoids assessment. PLoS One, 2019, 14(7): e0219944. https://doi.org/10.1371/journal.pone.0219944
Finisguerra, V., Dvorakova, T., Formenti, M., Van Meerbeeck, P., Mignion, L., Gallez, B., Van den Eynde, B. J. Metformin improves cancer immunotherapy by directly rescuing tumor-infiltrating CD8 T lymphocytes from hypoxia-induced immunosuppression. Journal for ImmunoTherapy of Cancer, 2023, 11(5): e005719. https://doi.org/10.1136/jitc-2022-005719
Conza, D., Mirra, P., Fiory, F., Insabato, L., Nicolò, A., Beguinot, F., Ulianich, L. Metformin: A new inhibitor of the Wnt signaling pathway in cancer. Cells, 2023, 12(17): 2182. https://doi.org/10.3390/cells12172182
Mamedov, M. R., Vedova, S., Freimer, J. W., Das Sahu, A., Ramesh, A., Arce, M. M., Meringa, A. D., Ota, M., Chen, P. A., Hanspers, K. et al. CRISPR screens decode cancer cell pathways that trigger γδ T cell detection. Nature, 2023, 621(7977): 188–195. https://doi.org/10.1038/s41586-023-06482-x
Menden, K., Marouf, M., Oller, S., Dalmia, A., Magruder, D. S., Kloiber, K., Heutink, P., Bonn, S. Deep learning–based cell composition analysis from tissue expression profiles. Science Advances, 2020, 6(30): eaba2619. https://doi.org/10.1126/sciadv.aba2619
Shi Y, Liu J, Li L, et al. Patient-derived skin tumor organoids with immune cells respond to metformin. Cell Organoid, 2024,https://doi.org/10.26599/CO.2024.9410001