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Pharmacological attenuation of melanoma by tryptanthrin pertains to the suppression of MITF-M through MEK/ERK signaling axis

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Abstract

Melanoma is the most aggressive among all types of skin cancers. The current strategies against melanoma utilize BRAFV600E, as a focal point for targeted therapy. However, therapy resistance developed in melanoma patients against the conventional anti-melanoma drugs hinders the ultimate benefits of targeted therapies. A major mechanism by which melanoma cells attain therapy resistance is via the activation of microphthalmia-associated transcription factor-M (MITF-M), the key transcription factor and oncogene aiding the survival of melanoma cells. We demonstrate that tryptanthrin (Tpn), an indole quinazoline alkaloid, which we isolated and characterized from Wrightia tinctoria, exhibits remarkable anti-tumor activity towards human melanoma through the down-regulation of MITF-M. Microarray analysis of Tpn-treated melanoma cells followed by a STRING protein association network analysis revealed that differential expression of genes in melanoma converges at MITF-M. Furthermore, in vitro and in vivo studies conducted using melanoma cells with differential MITF-M expression status, endogenously or ectopically, demonstrated that the anti-melanoma activity of Tpn is decisively contingent on its efficacy in down-regulating MITF-M expression. Tpn potentiates the degradation of MITF-M via the modulation of MEK1/2-ERK1/2-MITF-M signaling cascades. Murine models demonstrate the efficacy of Tpn in attenuating the migration and metastasis of melanoma cells, while remaining pharmacologically safe. In addition, Tpn suppresses the expression of mutated BRAFV600E and inhibits Casein Kinase 2α, a pro-survival enzyme that regulates ERK1/2 homeostasis in many tumor types, including melanoma. Together, we point to a promising anti-melanoma drug in Tpn, by virtue of its attributes to impede melanoma invasion and metastasis by attenuating MITF-M.

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Data availability

The data that support the findings of this study are available from the corresponding author on reasonable request.

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Acknowledgements

The Department of Biotechnology supported Lifetime imaging facility at RGCB is acknowledged for the development of genetically encoded cell probes and fluorescence imaging. The authors are grateful to Dr.T.R.Santosh Kumar, Scientist G, RGCB, and Dr. Shine V.J., Post-Doctoral Researcher, RGCB for the help rendered in conducting the mt-roGFP assay. The authors acknowledge the technical help rendered by Ms. Rayginia P.Tennyson, Ms. Keerthana.C.K. and Mr. Prakash. R. The authors thank the veterinarians, Dr. Archana.S, Dr. Arya Aravind, and Dr. Vishnu Sunil Jaikumar, and all the personnel of the Animal research facility for the help rendered during the animal experiments. This work was financially supported by research grants from the Department of Biotechnology, Government of India (Ruby John Anto) and the Council for Scientific and Industrial Research, Government of India (Anwar Shabna, Jayesh Antony, and Vijayakurup Vinod).

Funding

This work was financially supported by research grants from Department of Biotechnology, Government of India and Council for Scientific and Industrial Research, Government of India.

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Author notes

  1. Vijayasteltar B. Liju

    Present address: The Shraga Segal Department of Microbiology Immunology and Genetics, Faculty of Health Sciences, Ben-Gurion University of the Negev, 84105, Beer Sheva, Israel

  2. Sreekumar U. Aiswarya

    Present address: Department of Biotechnology, University of Calicut, Malappuram, Kerala, 673635, India

  3. Jayesh Antony

    Present address: Department of Zoology, St. Thomas College, Palai, Kottayam, Kerala, India

  4. Vinod Vijayakurup

    Present address: Department of Anatomy and Cell Biology, Cancer and Genetics Research Complex, University of Florida, Gainesville, FL, 32610, USA

  5. Anwar Shabna, Jayesh Antony and Vinod Vijayakurup have contributed equally to this work.

Authors and Affiliations

  1. Division of Cancer Research, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram, Kerala, 695014, India

    Anwar Shabna, Jayesh Antony, Vinod Vijayakurup, Minakshi Saikia, Vijayasteltar B. Liju, Archana P. Retnakumari, Vijai V. Alex, Mundanattu Swetha, Sreekumar U. Aiswarya, Somaraj Jannet & Ruby John Anto

  2. Department of Biotechnology and Microbiology, Thalassery Campus, Kannur University, Kannur, Kerala, 670661, India

    Nisthul A. Amrutha & Sadasivan Chittalakkottu

  3. KRIBS-BioNest, Third Campus of Rajiv Gandhi Centre for Biotechnology (RGCB) Kalamassery, Kochi, Kerala, India

    Uma Subramanian Unni

  4. Department of Pathology, Government Medical College, Kottayam, Kerala, 686008, India

    Sankar Sundaram

  5. Indian Institute of Information Technology and Management, Karyavattom, Kazhakkoottam, Kerala, 695581, India

    Daisy R. Sherin

  6. The Shraga Segal Department of Microbiology Immunology and Genetics, Faculty of Health Sciences, Ben-Gurion University of the Negev, 84105, Beer Sheva, Israel

    Nikhil Ponnoor Anto

  7. Department of Biotechnology, University of Calicut, Malappuram, Kerala, 673635, India

    Smitha V. Bava

  8. Department of Medical Microbiology, Immunology and Cell Biology, Southern Illinois University-School of Medicine, PO Box 19626, Springfield, IL, USA

    Sophia Ran

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  1. Anwar Shabna
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Contributions

RJA conceived and supervised the study. The design and methodology of the study was developed by RJA, JA, AS and VV. Material preparation, data collection and analysis were performed by AS, JA, VV, MS, VBL, RP, NA, CS, DR, SJ, AVV, MS, SUA, USU, SS and SR. Review and editing of the manuscript was performed by NPA, VV, JA and SVB. All authors commented on the previous versions of the manuscript. All authors have read and approved the final manuscript and have given consent for publishing the work.

Corresponding author

Correspondence to Ruby John Anto.

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Conflict of interest

The authors have no relevant financial or non-financial interests to disclose.

Ethical approval

All the animal studies were done in accordance with the protocols approved by the Institutional Animal Ethics Committee (IAEC), (IAEC/298/RUBY/2015, IAEC/299/RUBY/2015, IAEC/300/RUBY/2015, IAEC/192/RUBY/2012, IAEC/696/RUBY/2018) Rajiv Gandhi Centre for Biotechnology, Department of Biotechnology, Government of India.

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Not applicable as human subjects were not involved in the study.

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Shabna, A., Antony, J., Vijayakurup, V. et al. Pharmacological attenuation of melanoma by tryptanthrin pertains to the suppression of MITF-M through MEK/ERK signaling axis. Cell. Mol. Life Sci. 79, 478 (2022). https://doi.org/10.1007/s00018-022-04476-y

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