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The antimetastatic activity of orlistat is accompanied by an antitumoral immune response in mouse melanoma

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Abstract

Purpose

Fatty acid synthase (FASN), the multifunctional enzyme responsible for endogenous fatty acid synthesis, is highly expressed and associated with poor prognosis in several human cancers, including melanoma. Our group has previously shown that pharmacological inhibition of FASN with orlistat decreases proliferation, promotes apoptosis, and reduces the metastatic spread of B16-F10 cells in experimental models of melanoma. While most of the orlistat antitumor properties seem to be closely related to direct effects on malignant cells, its impact on the host immune system is still unknown.

Methods

The effects of orlistat on the phenotype and activation status of infiltrating leukocytes in primary tumors and metastatic lymph nodes were assessed using a model of spontaneous melanoma metastasis (B16-F10 cells/C57BL/6 mice). Cells from the primary tumors and lymph nodes were mechanically dissociated and immune cells phenotyped by flow cytometry. The expression of IL-12p35, IL-12p40, and inducible nitric oxide synthase (iNOS) was analyzed by qRT-PCR and production of nitrite (NO2−) evaluated in serum samples with the Griess method.

Results

Orlistat-treated mice exhibited a 25% reduction in the number of mediastinal lymph node metastases (mean 3.96 ± 0.78, 95% CI 3.63–4.28) compared to the controls (mean 5.7 ± 1.72; 95% CI 5.01–6.43). The drug elicited an antitumor immune response against experimental melanomas by increasing maturation of intratumoral dendritic cells (DC), stimulating the expression of cytotoxicity markers in CD8 T lymphocytes and natural killer (NK) cells, as well as reducing regulatory T cells (Tregs). Moreover, the orlistat-treatment increased serum levels of nitric oxide (NO) concentrations.

Conclusion

Taken together, these findings suggest that orlistat supports an antitumor response against experimental melanomas by increasing CD80/CD81-positive and IL-12-positive DC populations, granzyme b/NKG2D-positive NK populations, and perforin/granzyme b-positive CD8 T lymphocytes as well as reducing Tregs counts within experimental melanomas.

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Acknowledgements

The authors thank Fabiana Rossetto de Morais (Flow Cytometry Facility, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, São Paulo, Brazil) for the help with flow cytometry and data analysis. This work was supported by a grant from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP—2008/57471-7). LYA and FSM were supported by FAPESP fellowships (2012/25160-8 and 2010/52670-1, respectively). DCB was supported by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil (CAPES)—Finance code 001.

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Author notes

  1. Luciana Y. de Almeida and Flávia S. Mariano contributed equally to this work.

Authors and Affiliations

  1. Department of Oral Diagnosis, School of Dentistry of Piracicaba, State University of Campinas (UNICAMP), Piracicaba, São Paulo, Brazil. Av. Limeira 901, CP 52, Areão, Piracicaba, SP, 13414-903, Brazil

    Luciana Y. de Almeida, Flávia S. Mariano, Débora C. Bastos, Fernanda S. Moreira, Ricardo D. Coletta, Renata O. Mattos-Graner & Edgard Graner

  2. Urologic Oncology Program/Uro-Oncology Research Laboratories, Cedars-Sinai Medical Center, Samuel Oschin Comprehensive Cancer Institute, Los Angeles, CA, 90048, USA

    Karen A. Cavassani & Janna Raphelson

  3. Department of Basic and Applied Immunology, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Brazil

    Vânia S. Mariano

  4. Department of Oral Diagnosis and Pathology, School of Dentistry, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil

    Michelle Agostini

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  1. Luciana Y. de Almeida
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de Almeida, L.Y., Mariano, F.S., Bastos, D.C. et al. The antimetastatic activity of orlistat is accompanied by an antitumoral immune response in mouse melanoma. Cancer Chemother Pharmacol 85, 321–330 (2020). https://doi.org/10.1007/s00280-019-04010-1

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