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Research ArticleExperimental Studies

Peptide L13S Derived from the BRN2 POU Domain Reduces Metastasis In Vivo and Inhibits Cell Migration and Invasion In Vitro

MARIA CAROLINA MARIANO CESAR, RENATO ARRUDA MORTARA, VICTÓRIA SANTOS SOUZA, VITORIA RODRIGUES GUIMARAES ALVES, THAYSA PASCHOALIN, MARCO ANTONIO SOUFEN, GUILHERME MENDES DE FREITAS, FERNANDA FERNANDES MIRANDA DA CUNHA, DAYANE BATISTA TADA, ANDREY SENOS DOBROFF and DENISE COSTA ARRUDA
Anticancer Research January 2024, 44 (1) 71-84; DOI: https://doi.org/10.21873/anticanres.16789
MARIA CAROLINA MARIANO CESAR
1Núcleo Integrado de Biotecnologia, Universidade de Mogi das Cruzes, Mogi das Cruzes, SP, Brazil;
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RENATO ARRUDA MORTARA
2Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo, São Paulo, SP, Brazil;
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VICTÓRIA SANTOS SOUZA
1Núcleo Integrado de Biotecnologia, Universidade de Mogi das Cruzes, Mogi das Cruzes, SP, Brazil;
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VITORIA RODRIGUES GUIMARAES ALVES
1Núcleo Integrado de Biotecnologia, Universidade de Mogi das Cruzes, Mogi das Cruzes, SP, Brazil;
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THAYSA PASCHOALIN
3Departamento de Biofísica, Universidade Federal de São Paulo, São Paulo, SP, Brazil;
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MARCO ANTONIO SOUFEN
4Faculdade de Medicina da Universidade de Mogi das Cruzes, Universidade de Mogi das Cruzes, Mogi das Cruzes, SP, Brazil;
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GUILHERME MENDES DE FREITAS
1Núcleo Integrado de Biotecnologia, Universidade de Mogi das Cruzes, Mogi das Cruzes, SP, Brazil;
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FERNANDA FERNANDES MIRANDA DA CUNHA
1Núcleo Integrado de Biotecnologia, Universidade de Mogi das Cruzes, Mogi das Cruzes, SP, Brazil;
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DAYANE BATISTA TADA
5Instituto de Ciência e Tecnologia, Universidade Federal de São Paulo, São José dos Campos, SP, Brazil;
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ANDREY SENOS DOBROFF
6Division of Molecular Medicine, Department of Internal Medicine, University of New Mexico School of Medicine, Albuquerque, NM, U.S.A.;
7University of New Mexico Comprehensive Cancer Center, Albuquerque, NM, U.S.A.
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DENISE COSTA ARRUDA
1Núcleo Integrado de Biotecnologia, Universidade de Mogi das Cruzes, Mogi das Cruzes, SP, Brazil;
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  • For correspondence: denisearr@gmail.com
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Abstract

Background/Aim: The Brain-Specific Homeobox/POU Domain Protein 2 (BRN2) transcription factor supports melanoma progression by regulating the expression of several genes involved in cell migration and invasion. We hypothesized that a peptide designed based on the POU domain of BRN2 could block the BRN2 transcription activity and, consequently, reduce metastasis. Materials and Methods: Cell viability was accessed by Trypan Blue exclusion dye assay and xCelligence platform. Wound-healing scratch assay and transwell invasion with matrigel membrane assay were performed to analyze cell migration and invasion. The internalization mechanism of the L13S peptide was investigated using confocal microscopy and wound-healing scratch assay. The impact of L13S on cell protein expression was analyzed through western blotting. In vivo assays were conducted to evaluate the protective effect and toxicity of L13S in a metastatic model using murine melanoma cells. Results: Here, we show that the peptide named L13S can inhibit the migration and invasion of murine melanoma cells (B16F10-Nex2) as well as the migration of human melanoma cells (SK-MEL-25 and A375) by regulating the expression of proteins involved in motility. Mechanistically, we found that L13S is internalized by murine melanoma cells via macropinocytosis and binds actin filaments and nuclei. More importantly, in vivo studies indicated that the peptide was able to significantly inhibit lung metastasis in syngeneic models without off-target effects and with virtually no cytotoxicity toward normal organs. Conclusion: L13S peptide is a strong candidate for further development as an anticancer agent for the treatment of melanoma metastasis.

Key Words:
  • Peptide
  • BRN2
  • POU domain
  • melanoma
  • internalization
  • migration
  • invasion
  • metastatic model
  • Received November 7, 2023.
  • Revision received November 30, 2023.
  • Accepted December 4, 2023.
  • Copyright © 2024 International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.
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Peptide L13S Derived from the BRN2 POU Domain Reduces Metastasis In Vivo and Inhibits Cell Migration and Invasion In Vitro
MARIA CAROLINA MARIANO CESAR, RENATO ARRUDA MORTARA, VICTÓRIA SANTOS SOUZA, VITORIA RODRIGUES GUIMARAES ALVES, THAYSA PASCHOALIN, MARCO ANTONIO SOUFEN, GUILHERME MENDES DE FREITAS, FERNANDA FERNANDES MIRANDA DA CUNHA, DAYANE BATISTA TADA, ANDREY SENOS DOBROFF, DENISE COSTA ARRUDA
Anticancer Research Jan 2024, 44 (1) 71-84; DOI: 10.21873/anticanres.16789

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Peptide L13S Derived from the BRN2 POU Domain Reduces Metastasis In Vivo and Inhibits Cell Migration and Invasion In Vitro
MARIA CAROLINA MARIANO CESAR, RENATO ARRUDA MORTARA, VICTÓRIA SANTOS SOUZA, VITORIA RODRIGUES GUIMARAES ALVES, THAYSA PASCHOALIN, MARCO ANTONIO SOUFEN, GUILHERME MENDES DE FREITAS, FERNANDA FERNANDES MIRANDA DA CUNHA, DAYANE BATISTA TADA, ANDREY SENOS DOBROFF, DENISE COSTA ARRUDA
Anticancer Research Jan 2024, 44 (1) 71-84; DOI: 10.21873/anticanres.16789
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Keywords

  • peptide
  • BRN2
  • POU domain
  • Melanoma
  • internalization
  • migration
  • invasion
  • metastatic model
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