The structural conformation of the tachykinin domain drives the anti-tumoural activity of an octopus peptide in melanoma BRAFV600E

Br J Pharmacol. 2022 Oct;179(20):4878-4896. doi: 10.1111/bph.15923. Epub 2022 Jul 29.

Abstract

Background and purpose: Over past decades, targeted therapies and immunotherapy have improved survival and reduced the morbidity of patients with BRAF-mutated melanoma. However, drug resistance and relapse hinder overall success. Therefore, there is an urgent need for novel compounds with therapeutic efficacy against BRAF-melanoma. This prompted us to investigate the antiproliferative profile of a tachykinin-peptide from the Octopus kaurna, Octpep-1 in melanoma.

Experimental approach: We evaluated the cytotoxicity of Octpep-1 by MTT assay. Mechanistic insights on viability and cellular damage caused by Octpep-1 were gained via flow cytometry and bioenergetics. Structural and pharmacological characterization was conducted by molecular modelling, molecular biology, CRISPR/Cas9 technology, high-throughput mRNA and calcium flux analysis. In vivo efficacy was validated in two independent xerograph animal models (mice and zebrafish).

Key results: Octpep-1 selectively reduced the proliferative capacity of human melanoma BRAFV600E -mutated cells with minimal effects on fibroblasts. In melanoma-treated cells, Octpep-1 increased ROS with unaltered mitochondrial membrane potential and promoted non-mitochondrial and mitochondrial respiration with inefficient ATP coupling. Molecular modelling revealed that the cytotoxicity of Octpep-1 depends upon the α-helix and polyproline conformation in the C-terminal region of the peptide. A truncated form of the C-terminal end of Octpep-1 displayed enhanced potency and efficacy against melanoma. Octpep-1 reduced the progression of tumours in xenograft melanoma mice and zebrafish.

Conclusion and implications: We unravel the intrinsic anti-tumoural properties of a tachykinin peptide. This peptide mediates the selective cytotoxicity in BRAF-mutated melanoma in vitro and prevents tumour progression in vivo, providing a foundation for a therapy against melanoma.

Keywords: ROS; melanoma; metabolism; mitochondria; tachykinin-receptors.

MeSH terms

  • Adenosine Triphosphate
  • Animals
  • Antineoplastic Agents* / pharmacology
  • Calcium
  • Cell Line, Tumor
  • Humans
  • Melanoma* / drug therapy
  • Melanoma* / pathology
  • Mice
  • Mutation
  • Octopodiformes / chemistry
  • Peptides / pharmacology
  • Proto-Oncogene Proteins B-raf / genetics
  • Proto-Oncogene Proteins B-raf / therapeutic use
  • RNA, Messenger
  • Reactive Oxygen Species
  • Tachykinins / genetics
  • Tachykinins / therapeutic use
  • Zebrafish / genetics

Substances

  • Antineoplastic Agents
  • Peptides
  • RNA, Messenger
  • Reactive Oxygen Species
  • Tachykinins
  • Adenosine Triphosphate
  • BRAF protein, human
  • Proto-Oncogene Proteins B-raf
  • Calcium