Twist1-induced epithelial dissemination requires PRKD1 signaling

Dan Georgess, Veena Padmanaban, Orit Katarina Sirka, Kester Coutinho, Alex Choi, Gabriela Frid, Neil M. Neumann, Takanari Inoue, Andrew J. Ewald

Research output: Contribution to journalArticle

Abstract

Dissemination is an essential early step in metastasis but its molecular basis remains incompletely understood. To define the essential targetable effectors of this process, we developed a 3D mammary epithelial culture model, in which dissemination is induced by overexpression of the transcription factor Twist1. Transcriptomic analysis and ChIP-PCR together demonstrated that protein kinase D1 (Prkd1) is a direct transcriptional target of Twist1 and is not expressed in the normal mammary epithelium. Pharmacologic and genetic inhibition of Prkd1 in the Twist1-induced dissemination model demonstrated that Prkd1 was required for cells to initiate extracellular matrix (ECM)-directed protrusions, release from the epithelium, and migrate through the ECM. Antibody-based protein profiling revealed that Prkd1 induced broad phosphorylation changes, including an inactivating phosphorylation of β-catenin and two microtubule depolymerizing phosphorylations of Tau, potentially explaining the release of cell-cell contacts and persistent activation of Prkd1. In patients with breast cancer, TWIST1 and PRKD1 expression correlated with metastatic recurrence, particularly in basal breast cancer. Prkd1 knockdown was sufficient to block dissemination of both murine and human mammary tumor organoids. Finally, Prkd1 knockdown in vivo blocked primary tumor invasion and distant metastasis in a mouse model of basal breast cancer. Collectively, these data identify Prkd1 as a novel and targetable signaling node downstream of Twist1 that is required for epithelial invasion and dissemination. Significance: Twist1 is a known regulator of metastatic cell behaviors but not directly targetable. This study provides a molecular explanation for how Twist1-induced dissemination works and demonstrates that it can be targeted.

Original languageEnglish (US)
Pages (from-to)204-218
Number of pages15
JournalCancer Research
Volume80
Issue number2
DOIs
StatePublished - Jan 15 2020

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Protein Kinases
Breast Neoplasms
Phosphorylation
Extracellular Matrix
Breast
Epithelium
Organoids
Neoplasm Metastasis
Catenins
Microtubules
Transcription Factors
Recurrence
Polymerase Chain Reaction
Antibodies
Neoplasms
Proteins

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Georgess, D., Padmanaban, V., Sirka, O. K., Coutinho, K., Choi, A., Frid, G., ... Ewald, A. J. (2020). Twist1-induced epithelial dissemination requires PRKD1 signaling. Cancer Research, 80(2), 204-218. https://doi.org/10.1158/0008-5472.CAN-18-3241

Twist1-induced epithelial dissemination requires PRKD1 signaling. / Georgess, Dan; Padmanaban, Veena; Sirka, Orit Katarina; Coutinho, Kester; Choi, Alex; Frid, Gabriela; Neumann, Neil M.; Inoue, Takanari; Ewald, Andrew J.

In: Cancer Research, Vol. 80, No. 2, 15.01.2020, p. 204-218.

Research output: Contribution to journalArticle

Georgess, D, Padmanaban, V, Sirka, OK, Coutinho, K, Choi, A, Frid, G, Neumann, NM, Inoue, T & Ewald, AJ 2020, 'Twist1-induced epithelial dissemination requires PRKD1 signaling', Cancer Research, vol. 80, no. 2, pp. 204-218. https://doi.org/10.1158/0008-5472.CAN-18-3241
Georgess D, Padmanaban V, Sirka OK, Coutinho K, Choi A, Frid G et al. Twist1-induced epithelial dissemination requires PRKD1 signaling. Cancer Research. 2020 Jan 15;80(2):204-218. https://doi.org/10.1158/0008-5472.CAN-18-3241
Georgess, Dan ; Padmanaban, Veena ; Sirka, Orit Katarina ; Coutinho, Kester ; Choi, Alex ; Frid, Gabriela ; Neumann, Neil M. ; Inoue, Takanari ; Ewald, Andrew J. / Twist1-induced epithelial dissemination requires PRKD1 signaling. In: Cancer Research. 2020 ; Vol. 80, No. 2. pp. 204-218.
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