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Flotillin-1 palmitoylation is essential for its stability and subsequent tumor promoting capabilities

Oncogene volume 43, pages 1063–1074 (2024)Cite this article

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Abstract

Flotillin-1 contributes to invasion and metastasis in triple negative breast cancer (TNBC) and is modified post-translationally through palmitoylation. Palmitoylation, the process of conjugating palmitoyl-CoA to proteins, plays an essential role in protein stability and trafficking. Thus far, there has not been any investigation into the role of flotillin-1 palmitoylation in the context of metastasis in vivo. To address the role of flotillin-1 palmitoylation in metastasis, MDA-MB-231 cells expressing palmitoylation defective flotillin-1 constructs were used as models. Compared to flotillin-1 WT expressing tumors, flotillin-1 palmitoylation defective displayed abrogated tumor progression and lung metastasis in vivo in both spontaneous and experimental models. Further mechanistic investigation led to the identification of zDHHC5 as the main palmitoyl acyltransferase responsible for palmitoylating endogenous flotillin-1. Modulation of flotillin-1 palmitoylation status through mutagenesis, zDHHC5 silencing, and 2-bromopalmitate inhibition all resulted in the proteasomal degradation of flotillin-1 protein. To assess if flotillin-1 palmitoylation can be inhibited for potential clinical relevance, we designed a competitive peptide fused to a cell penetrating peptide sequence, which displayed efficacy in blocking flotillin-1 palmitoylation in vitro without altering palmitoylation of other zDHHC5 substrates, highlighting its specificity. Additionally, TNBC xenograft tumor models expressing a doxycycline inducible flotillin-1 palmitoylation inhibiting peptide displayed attenuated tumor growth and lung metastasis. Collectively, these results reveal a novel palmitoylation dependent mechanism which is essential for the stability of flotillin-1 protein. More specifically, disruption of flotillin-1 palmitoylation through mutagenesis or competitive peptide promoted flotillin-1 protein degradation, subsequently impeding its tumor promoting and metastasis-inducing effects in TNBC tumor models.

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Fig. 1: Palmitoylation-defective flotillin-1 displays altered stability and localization in TNBC cell lines.
Fig. 2: zDHHC5 contributes to the stability of endogenous flotillin-1 by preventing its proteasomal degradation.
Fig. 3: Flotillin-1 palmitoylation defective TNBC cells display attenuated tumor growth and lung metastasis.
Fig. 4: Targeting flotillin-1 palmitoylation with a competitive peptide attenuates tumor growth and lung metastasis.

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Acknowledgements

We would like to thank Dr. Brittany Harlow for offering her support during data collection as well as the UT Austin flow cytometry core for their support with the FACS experimentation.

Funding

University of Texas Vice President of Research Creative Grant.

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Authors and Affiliations

  1. Department of Nutritional Sciences, The University of Texas at Austin, Austin, TX, USA

    Bryan McClellan, Crystal N. Wilson, Christopher A. Jolly & Linda deGraffenried

  2. Department of Molecular Biosciences, The University of Texas at Austin, Austin, TX, USA

    Crystal N. Wilson

  3. Mays Cancer Center, University of Texas Health Science Center, San Antonio, TX, USA

    Andrew J. Brenner, Christopher A. Jolly & Linda deGraffenried

  4. Division of Hematology and Oncology, Department of Medicine, University of Texas Health Science Center, San Antonio, TX, USA

    Andrew J. Brenner

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BM designed and performed all experiments unless stated otherwise and wrote the manuscript, CW assisted in animal experiments and edited the manuscript, AB edited the manuscript and provided supervision, CJ edited the manuscript and provided supervision, LD edited the manuscript, provided supervision, and acquired funding for the study.

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Correspondence to Linda deGraffenried.

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McClellan, B., Wilson, C.N., Brenner, A.J. et al. Flotillin-1 palmitoylation is essential for its stability and subsequent tumor promoting capabilities. Oncogene 43, 1063–1074 (2024). https://doi.org/10.1038/s41388-024-02946-0

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