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A novel Pyk2-derived peptide inhibits invadopodia-mediated breast cancer metastasis

Oncogene volume 42, pages 278–292 (2023)Cite this article

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A Correction to this article was published on 09 December 2022

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Abstract

Dissemination of cancer cells from the primary tumor into distant body tissues and organs is the leading cause of death in cancer patients. While most clinical strategies aim to reduce or impede the growth of the primary tumor, no treatment to eradicate metastatic cancer exists at present. Metastasis is mediated by feet-like cytoskeletal structures called invadopodia which allow cells to penetrate through the basement membrane and intravasate into blood vessels during their spread to distant tissues and organs. The non-receptor tyrosine kinase Pyk2 is highly expressed in breast cancer, where it mediates invadopodia formation and function via interaction with the actin-nucleation-promoting factor cortactin. Here, we designed a cell-permeable peptide inhibitor that contains the second proline-rich region (PRR2) sequence of Pyk2, which binds to the SH3 domain of cortactin and inhibits the interaction between Pyk2 and cortactin in invadopodia. The Pyk2-PRR2 peptide blocks spontaneous lung metastasis in immune-competent mice by inhibiting cortactin tyrosine phosphorylation and actin polymerization-mediated maturation and activation of invadopodia, leading to reduced MMP-dependent tumor cell invasiveness. The native structure of the Pyk2-PRR2:cortactin-SH3 complex was determined using nuclear magnetic resonance (NMR), revealing an extended class II interaction surface spanning the canonical binding groove and a second hydrophobic surface which significantly contributes to ligand affinity. Using structure-guided design, we created a mutant peptide lacking critical residues involved in binding that failed to inhibit invadopodia maturation and function and consequent metastatic dissemination in mice. Our findings shed light on the specific molecular interactions between Pyk2 and cortactin and may lead to the development of novel strategies for preventing dissemination of primary breast tumors predicted at the time of diagnosis to be highly metastatic, and of secondary tumors that have already spread to other parts of the body.

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Fig. 1: Increased expression of Pyk2 and cortactin is associated with breast cancer metastasis and poor patient prognosis.
Fig. 2: A cell-permeable peptide derived from the second proline-rich region of Pyk2 blocks spontaneous lung metastasis in an immune-competent xenograft mouse model.
Fig. 3: Pyk2-PRR2 suppresses MMP-dependent 3D invasiveness of breast tumor cells.
Fig. 4: Pyk2-PRR2 peptide inhibits Pyk2-cortactin interactions in invadopodia and consequent invadopodium maturation and function in breast tumor cells.
Fig. 5: Pyk2-PRR2 inhibits cortactin tyrosine phosphorylation and consequent barbed end formation in invadopodia.
Fig. 6: Structure of the cortactin-SH3:Pyk2-PRR2 complex.
Fig. 7: A Pyk2-PRR2 super-mutant peptide does not inhibit the interaction between Pyk2 and cortactin at invadopodia.
Fig. 8: The Pyk2 super-mutant peptide does not block spontaneous lung metastasis in a xenograft mouse model.

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Acknowledgements

This paper is dedicated to the memory of Mr. Abd Elhaleem Abu Salah, who sadly passed away before completion of this work. We wish to thank Dr. Avraham Samson, Dr. Moshe Dessau, and Ms. Trishna Saha for technical assistance and advice, Mr. Jonathan Solomon and Ms. Michal Gendler for critical reading of the manuscript, and Ms. Natalia Saleev and Ms. Michal Gendler for technical assistance during revisions. This work was funded by the Israel Cancer Research Fund (grant number 20-101-PG), the Israel Cancer Association (grant number 20210071) and the Israel Science Foundation (grant number 2142/21) (to HG-H), and the Israel Science Foundation (grant number 964/19) (to JC).

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Author notes

  1. These authors contributed equally: Shams Twafra, Chana G. Sokolik.

  2. These authors jointly supervised this work: Tal Sneh, Kolluru D. Srikanth.

Authors and Affiliations

  1. Cell Migration and Invasion Laboratory, The Azrieli Faculty of Medicine, Bar-Ilan University, Safed, 1311502, Israel

    Shams Twafra, Tal Sneh, Kolluru D. Srikanth, Tomer Meirson, Alessandro Genna & Hava Gil-Henn

  2. Bio-NMR Laboratory, Department of Chemistry, Bar-Ilan University, Ramat-Gan, 52900, Israel

    Chana G. Sokolik & Jordan H. Chill

  3. Davidoff Cancer Center, Rabin Medical Center-Beilinson Hospital, Petah Tikva, Israel

    Tomer Meirson

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Contributions

Conceptualization: JHC, HG-H; formal analysis: ST, CGS, TM; funding acquisition: JHC, HG-H; investigation: ST, CGS, TS, KDS, AG; supervision: JHC, HG-H; writing - original draft: JHC, HG-H; writing - review and editing: ST, CGS, TS, KDS, AG, JHC, HG-H.

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Correspondence to Jordan H. Chill or Hava Gil-Henn.

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Twafra, S., Sokolik, C.G., Sneh, T. et al. A novel Pyk2-derived peptide inhibits invadopodia-mediated breast cancer metastasis. Oncogene 42, 278–292 (2023). https://doi.org/10.1038/s41388-022-02481-w

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